Binding molecules, chimeric antigen receptors, vaccine preparations, and methods for their production.

Abstract

The invention relates to a binding molecule possessing one or more (preferably all) highly specific binding levels to the human cytomegalovirus (HCMV) US28 protein, very low nonspecific binding levels to healthy (uninfected) cells, and / or unknown binding capacity of the variant, as well as the nucleic acid molecules encoding the aforementioned binding molecules. The binding molecules are designed to bind to the extracellular domain 3 (ECD3) of the human cytomegalovirus (HCMV) US28 protein, the third of four extracellular domains represented by US28, corresponding to positions 167 to 183 of the US28 protein sequence as determined by SEQ ID NO:5. The binding molecules of the invention have been shown to possess superior binding properties, including specific binding specificity for metastatic and / or invasive HCMV-infected cancers, including breast cancers.According to several preferred schemes, the binding molecule is selected from antibodies (including, for example, BiTE antibodies) and chimeric antigen receptors (CARs), or their functional variants, fragments, fusion proteins, and / or conjugates. The invention also relates to cells expressing the aforementioned binding molecules, such as CAR-expressing cells, including CAR-T cells, CAR-NK cells, and CAR-M cells.

Description

[0001] Therapeutic and Diagnostic Agents and Uses Thereof

[0002] FIELD OF INVENTION

[0003] The present invention relates to the field of virology. More specifically, the invention relates to therapeutic and diagnostic agents targeting a specific region of the US28 protein, as encoded by human cytomegalovirus (HCMV), and therapies related thereto including but not limited to HCMV-infected cancers and other conditions associated with latent or lytic HCMV infections.

[0004] REFERENCES

[0005] The listing or discussion of an apparently prior-published document in this specification should not necessarily be taken as an acknowledgement that the document is part of the state of the art or is common general knowledge. The references disclosed, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein by reference.

[0006] BACKGROUND

[0007] Human cytomegalovirus (HCMV), also known as human herpes Virus 5 (HHV- 5), is a ubiquitous, opportunistic DNA virus carried by 56-94% of the population worldwide (Geisler et al, Cancers, 2019, 11 : 1842; Zuhair et al, Rev Med Virol, 2019. 29(3) : p. e2034).

[0008] In most immunocompetent individuals, HCMV infections are asymptomatic, remain undiagnosed and are considered harmless, since viral replication is well- controlled by the host immune system (Boeckh and Geballe, J Clin Invest, 2011. 121(5): p. 1673-80).

[0009] Similar to all herpesviruses, after primary infection, HCMV establishes a lifelong persistence as a latent infection. The latent infection is characterized by low-level or non-existent virus replication with the viral genome residing predominantly in the CD34+hematopoietic progenitor cell population residing in the bone marrow (Collins- McMillen et al., Viruses, 2018. 10(8)).

[0010] It is assumed that latent HCMV may intermittently reactivate in a stochastic manner unless continuously controlled by the host immune system. For this reason, the virus may cause complications in certain circumstances, for example in immunocompromised patients, in whom not only primary HCMV infection, but also reinfection or reactivation can cause a life-threatening disease that affects many organs causing considerable morbidity and mortality (Boeckh and Geballe, supra; Griffiths et al, J Pathol, 2015. 235(2) : p. 288-97). HCMV is one of the most common congenital viral infections and most important cause of birth defects (Davis et a / , Birth Defects Res, 2017. 109(5) : p. 336-346). It is becoming increasingly clear that HCMV infection over the life-course may also play a role in the pathogenesis of atherosclerosis, autoimmune diseases, and several malignancies, particularly glioblastoma multiforme (Soderberg-Naucler, J Intern Med, 2006, 259(3) : p. 219-46; Cobbs, Curr Opin Virol, 2019. 39: p. 49-59). HCMV serostatus may additionally impact the clinical course of burns, trauma, and sepsis (Soderberg-Naucler, 2006, supra; Limaye, et al, JAMA, 2008. 300(4) : p. 413-22; Osawa Singh, Crit Care, 2009. 13(3): p. R68).

[0011] Latent HCMV infection can be reactivated during an inflammatory process when the progenitor cells differentiate into monocyte / infiltrating macrophages or dendritic cells (DCs), and these cells can disseminate the virus to peripheral organs (Soderberg- Naucler et al, Cell, 1997, 91: 119-126). Reactivated HCMV, carried by these inflammatory cells, can reach all body tissues, and infect and replicate in a broad number of cell types (Ljungman et al., Infect. Dis. Clin. North. Am., 2010, 24: 319- 337). The infection is further transmitted by all body fluids, including saliva and breast milk (Hamprecht et al., Lancet, 2001, 357: 513-518). Ninety percent of breast milk samples from HCMV seropositive women contain the virus, and that results in about 30% HCMV prevalence in children at one year of age. Nursing and parental contact, therefore, constitutes an important route to acquiring the HCMV infection in early infancy or childhood (Hamprecht et al., 2001, supra).

[0012] The Cytomegalovirus Genome and Genetic Diversity:

[0013] As discussed in Berg et al, 2019, PLoS ONE 14(9): e0222053, the CMV genome consists of monopartite, linear, double-stranded DNA and is roughly 235 kb in size. It contains more than 750 translated ORFs (Stern-Ginossar et al., Science, 2012, 338(6110) : 1088-93) which can be divided into two regions - the unique long (UL) and unique short (US) regions - flanked by terminal and internal inverted repeats.

[0014] Cytomegalovirus has adapted a wide range of strategies to avoid immune detection and facilitate dissemination of infection. These strategies are based on manipulation and modulation of the host's immune response during infection, e.g. by expression of virally encoded homologs of receptors and ligands important for the normal function of the human immune system. By encoding a 2 to 3-fold greater number of gene products than other human herpesviruses, many of which have been shown to interact with and manipulate the human immune system (Mocarski, Trends Microbiol., 2002; 10(7) :332-9), CMV has an unparalleled number of tools available for modifying the host's immune response. The genetic variation between circulating CMV strains is large and a recent study reported that 75% of the strains contain disruptive mutations and polymorphisms in several genes (Sijmons eta / ., J. Virol., 2015, 89(15) : 7673-7695). In order to exclude disruptive mutations due to serial passage, the authors of the study only used strains passaged 1-2 times and verified most of the observed mutations directly from clinical samples. For the genes UL40 and UL111A, mutations causing functional knockouts were found in 9.9% and 5.5% of the investigated strains, respectively (Sijmons et al., 2015, supra). UL111A is a functional interleukin-10 homolog that can inhibit a normal immune response (Mocarski, 2002, supra; Engel and Ang u Io, Adv Exp Med Biol., 2012, 738:256-76). The signal peptide of UL40 facilitates surface expression of HLA-E on infected cells, which is a ligand for a natural killer cell inhibitory receptor (Wilkinson et al., J Clin Virol., 2008; 41(3) :206-12).

[0015] Other CMV genes that are also highly variable are the chemokine homolog UL146 where 14 distinct genotypes have been identified (Dolan et al., J Gen Virol., 2004, 85(Pt 5) : 1301-12), and the chemokine scavenging receptor (Kledal et al., FEBS Lett., 1998; 441(2): 209-14) and G protein-coupled receptor US28 where numerous N-terminal polymorphisms have been reported (Goffard et al., virus Genes, 2006; 33(2) : 175-81; Arav-Boger et al., J Infect Dis., 2002; 186(8) : 1057-64).

[0016] Berg et al, 2019 (supra) reports that this degree of genetic diversity is not observed for other human herpesviruses (Sijmons et al., 2015, supra) and poses the question of why CMV exerts such variability among important immunomodulatory genes and how it affects the virus-host interaction.

[0017] It is considered that a large part of the HCMV pathogenesis is associated with viral latency, which is closely linked to virus ability to escape from the humoral and cellular host immune responses through a number of mechanisms (Manandhar et al., Int J Mol Sci, 2019. 20(15)). One of the most important of such mechanisms include this high, ever-changing genetic diversity of the HCMV. The HCMV genome varies between different individuals and even within the same host (Gorzer et al., J Virol, 2010, 84(14) : 7195-7203; Renzette et al., PLoS Pathog, 2011, 7(5) : el001344; Renzette et al., Curr Opin Virol, 2014. 8: 109-15; Renzette et al., Proc Natl Acad Sci USA, 2015, 112(30) : E4120-8; Renzette et al., J Virol, 2017, 91(5)). New host infections give rise to a unique viral strain for each infected individual and generate selection events where a new genotype becomes dominant due to the selective pressure of the immune response (Renzette et al., 2011, supra). It is possible that both viral and host factors can contribute to fostering viral genetic drift during the HCMV infection (Vabret et al., Trends Immunol, 2017, 38(1) : 53-65; Christensen & Paludan, Cell Mol Immunol, 2017, 14(1) : 4-13). In addition, each patient is likely to be infected with multiple CMV strains as previously extensively reported in the literature (Renzette et al., 2015, supra). The presence of multiple strains in the same individual enable recombination from the different HCMV strains. Recombination is considered to stand out as a major driver of HCMV genetic diversity (Suarez et al., J Infect Dis, 2019, 220(5) : 781-791; Sijmons et al., 2015, supra; Lassalle et al., Virus Evol, 2016, 2(1): vew017; Cudini et al., Proc Natl Acad Sci USA, 2019, 116(12) : 5693- 5698). Reasserting the highly diverse regions would create new combinations to ensure efficient immune evasion, which also can impact the pathogenicity of the virus. Consistently, mixed HCMV infection has been associated with poor clinical outcome in immunocompromised individuals in several studies (Coaquette et al., Clin Infect Dis, 2004, 39(2) : 155-161; Lisboa et al., Transpl Infect Dis, 2012, 14(2): 132-140; Houldcroft et al., Front Microbiol, 2016, 7: 1317).

[0018] HCMV diversity is moreover driven by genetic polymorphisms, which are not evenly distributed across the genome (Sijmons et al., 2015, supra). Selection is stronger in protein regions exposed on the virion surface and for viral proteins expressed at the host cell membrane in the extracellular domains (Mozzi et al., PLoS Pathog, 2020, 16(5) : el008476). The selective pressure exerted by the host immune system has likely played a major role in the shaping of genetic diversity among circulating HCMV strains. Thus, several sites targeted by positive selection are located within epitopes recognized by human antibodies or in protein regions that directly interact with host molecules involved in immune response (Sijmons eta / ., 2015, supra; Mozzi et al., 2020, supra). These features are consistent with an ongoing hide and seek interplay between HCMV and the human immune system.

[0019] The strain variability and constantly mutating virus make both viral diagnostics and the vaccine and drug development demanding against the HCMV and set limits for the current antiviral drug treatment. Vaccine development against HCMV has over many years been of high priority for the medical community, but no effective vaccines have so far been approved against HCMV.

[0020] Oncogenic Properties of HCMV

[0021] HCMV encoded proteins display diverse oncogenic functions (Geisler et al, 2019, supra). Upon entry into the host cell, tegument proteins of the HCMV virion, such as pUL48, are released, disabling cellular intrinsic and innate immune responses, and promoting enhanced metabolic activity of the host cells (Kumari et al. , Cell Death Dis., 2017, 8: e3078). These HCMV-encoded proteins may enable the cells to surpass the Gl-phase to facilitate rapid cell division (Kumari et al., 2017, supra). Through upregulation of anti-apoptotic genes and downregulation of pro-apoptotic genes, cells enter a state of enhanced survival. After the entry of viral DNA into the cell nucleus, cellular RNA polymerases I and II (Pol I and II) are employed to transcribe the viral genes by binding to the major immediate early promoter (MIEP) (Kostopoulou et al., Oncotarget, 2017, 8: 96536- 96552). The first genes that are expressed are the immediate early (IE) genes. The IE proteins derived from such genes act as transcription factors controlling both early and late viral gene expression, and direct host gene expression. Such proteins are necessary to establish lytic infection and are crucial for viral reactivation from latency (Kumari et al., 2017, supra; Tamrakar et a / ., J. Virol., 2005, 79: 15477-15493). Lytic HCMV infection leads to a dysregulated cell cycle, and the IE gene products interfere with key cellular factors, including retinoblastoma protein family (Rb), cyclins, p53, Wnt, phosphatidylinositol 3-kinase / Akt, human telomerase reverse transcriptase (hTERT), and N F-KB to increase the immortal properties of infected cells (Moussawi et al., Sci. Rep., 2018, 8: 12574). These pathways are commonly activated in cancer cells. Activation of mitogenic signals, delivered by proto-oncogenes such as Fos and Myc, can be induced by IE proteins in HCMV infected cells (Hagemeier et al., J. Virol., 1992, 66: 4452-4456). Moreover, the MYB gene is induced in HCMV infected cells resembling the enhanced MYB gene expression in HPV-related carcinoma (Moussawi et al., 2018, supra). In addition to the mitogenic signals, HCMV infection causes chromosomal aberrations through deterioration of DNA repair pathways, resulting in genetic instability in the infected cells (Straat et al., J. Natl. Cancer Inst., 2009, 101 : 488-497; Siew et al., J. Biomed. Sci., 2009, 16: 107). This drives the development of genetic mutations.

[0022] Various HCMV-encoded, G-protein-coupled-receptor (GPCR)-like proteins, including US27, US28, UL33, and UL78, have been reported to display important oncogenic functions (Heukers et al., Oncogene, 2018, 37: 4110-4121). G-proteins activate both metabolic and oncogenic key signaling pathways, such as cAMP and the PI3K signaling pathways, of which the latter is critical for the emergence of anchorageindependent growth and oncogenic transformation of epithelial cells (Moussawi et al., 2018, supra; Boroughs et al., Nat. Cell Biol., 2015, 17: 351-359). The HCMV-2.7 early gene transcript is a long non-coding (Inc) RNA that interacts directly with complex I of the respiratory chain in mitochondria, preventing mitochondria-induced cell death by inhibiting Fas-ligand interactions and granzyme B by binding to caspase 8, improving the oxidative capacity and maintaining energy production in the infected cells (Reeves et al., Science, 2007, 316: 1345-1348).

[0023] HCMV has also developed several ways to manipulate the innate and adaptive immune responses to decrease its immune surveillance and improve its chances of surviving in its immunocompetent host, which may well account for the important immune evasive mechanisms in the HCMV-infected cancer cells. HCMV encodes multiple proteins that modulate NK cell recognition of the infected cells (Fielding et al., PLoS Pathog., 2014, 10: el004058), and increase CD8+T cell tolerability for the viral proteins. HCMV encoded proteins can stimulate the development of an immature phenotype of DC, which reduces the activation of CD4+T cell responses (Wagner et al., J. Leukoc Biol., 2008, 83: 56-63), and additionally, decreases the elimination of infected cells by CD8+cytotoxic T cells.

[0024] HCMV therapeutic targets:

[0025] Currently, the only antiviral therapy for HCMV available relies on nucleoside analogs, such as ganciclovir (GCV) and valganciclovir (VAL-GCV) (Rawlinson et al., Lancet Infect Dis, 2017, 17(6): el77-el88; James & Kimberlin, Curr Opin Pediatr, 2016, 28(1) : 81-85), which have several disadvantages such as poor bioavailability, toxic side-effects and the risk of developing drug resistance. Current results indicate that DNA polymerase (UL54) and viral phosphotransferase (UL97), two highly polymorphic HCMV genes, play important role in drug resistance against GCV (Komatsu et al., Antiviral Res, 2014, 101 : 12-25).

[0026] In addition, importantly, the existing antivirals can only be used to treat lytic HCMV infections, but cannot clear the latent virus. Eradication or reducing the latent reservoir would be a favorable way to reduce the burden of HCMV related diseases in several patient groups.

[0027] The previously developed anti-HCMV drugs, such as ganciclovir (GCV), foscarnet (FOS), and cidofovir (CDV), all target the UL54 viral DNA polymerase. Yet, antiviral toxicity and HCMV antiviral drug resistance constitute a growing therapeutic challenge in the transplant setting and so new anti-HCMV drugs with novel viral targets are highly needed (Burrel et al, 2014, Lack of influence of human cytomegalovirus (HCMV) susceptibility to current antiviral drugs on HCMV-encoded US28 chemokine receptor polymorphism, Poster presentation at ECCMID 2014, Barcelona).

[0028] Burrel et al, 2014 (supra) taught that, because of its potential roles in viral dissemination and persistence as well as in smooth muscle cell migration and tumorigenesis, HCMV-encoded US28 constitutes a potential target for novel antiviral therapies.

[0029] HCMV US28 is a seven transmembrane protein belonging to a class of G-protein coupled receptors (GCPRs). GCPRs constitute the largest family of proteins targeted by approved drugs (Sriram & Insel, Mol Pharmacol, 2018. 93(4) : 251-258), and share common architecture, each consisting of a single polypeptide with an extracellular N- terminus, an intracellular C-terminus and seven hydrophobic transmembrane domains (TM1-TM7) linked by three extracellular loops (ECL1-3) (Alexander et al., Br J Pharmacol, 2019, 176 Suppl 1 : S21-S141). The full sequence of US28 as encoded by HCMV strain DB (Accession number KT959235) is provided in the present application as SEQ ID NO: 5, wherein:

[0030] - the N-terminal extracellular domain (also referred to herein as ECD1 as it is the first extracellular domain) is provided herein as SEQ ID NO: 1 and corresponds to positions 1-37 of SEQ ID NO:5,

[0031] - the first extracellular loop (ECL1; although also referred to herein as ECD2 as it is the second extracellular domain) is provided herein as SEQ ID NO: 2 and corresponds to positions 91-101 of SEQ ID NO: 5,

[0032] - the second extracellular loop (ECL2; although also referred to herein as ECD3 as it is the third extracellular domain) is provided herein as SEQ ID NO: 3 and corresponds to positions 167-183 of SEQ ID NO: 5; and

[0033] - the third extracellular loop (ECL3; although also referred to herein as ECD4 as it is the fourth extracellular domain) is provided herein as SEQ ID NO: 4 and corresponds to positions 250-273 of SEQ ID NO: 5.

[0034] Burrel et al (supra) assessed the levels of polymorphism in the US28 protein amongst HCMV clinical strains, and concluded that the level of polymorphisms for US28 amongst clinical strains is higher than the level of polymorphisms previously reported for other HCMV-encoded proteins, such as UL97 phosphotransferase and UL44 processivity factor, although this polymorphism does not significantly vary according to HCMV susceptibility or resistance to currently approved antiviral drugs (i.e., GCV, FOS, and CDV), supporting therefore the idea that HCMV encoded US28 chemokine receptor may constitute a promising viral target for anti-HCMV drugs, especially in case of HCMV resistance.

[0035] A US28-focussed approach was taken by the authors of WO 2019 / 151865, as also reported in the equivalent journal article De Groof eta / , 2019, Mol. Pharmaceutics, 16: 3145-3156. The authors reported that they had generated single heavy chain variable domain antibodies (VHH), exemplified by a particular VHH referred to as VUN100 (SEQ ID NO: 60 of the present application), that was said to specifically detect US28 in glioblastoma (GBM) tissues and inhibit ligand-dependent and constitutive US28 activity, and which the authors reported to consequently impair US28-dependent GBM growth in vitro and in vivo in an orthotopic xenograft model.

[0036] VUN 100 was shown to bind to a discontinuous epitope, which comprise multiple binding positions in the N-terminal extracellular region of US28 (positions 1-37, also referred to herein as ECD1), and further influenced by the presence of the third extracellular loop (ECL3, positions 250-273, also referred to herein as ECD4) of US28, as discussed in Example 3 of WO 2019 / 151865 (page 36, lines 11-32) and the legend to Fig 2 of De Groof et al, 2019 (supra). An assay to compare the binding of the VUN 100 Ab to US28-expressing HEK293T membranes versus to mock transfected HEK293T membranes showed that 20% of the mock transfected cells were bound by VUN100, and it only achieved a relative specificity score of 4 for the US28-expressing HEK293T membranes (De Groof et al., 2019 (supra) in their supporting information, Figure SI. A thereof, and Table 2 of the present application). The apparent ability to distinguish between US28- expressing and US28-negative cells, with a specificity score of only around 4, is potentially sub-optimal, and raises concerns about the ability of VUN100 to provide specifically targeted effects to HCMV-infected cells, whilst avoiding unacceptable levels of off-target side effects in healthy cells. It also raises concerns about the ability of VUN100 to be useful in the context of reliably identifying US28-expressing cells (in particular, HCMV-infected cells) in assays, including diagnostic assays, such as for use in immunohistochemistry (IHC). Furthermore, in a further publication, it has been acknowledged that the potency of US28-targeting nanobodies needs to be validated in a viral and potentially in vivo setting (De Groof et al., 2021, Pharmacol Rev 73:828- 846).

[0037] The provision of binding molecules having a substantially greater ability than VUN100 to bind specifically to US28-expressing cells (in particular, HCMV-infected cells) and / or to minimize off-target binding to cells that do not express US28 (in particular, cells that are not HCMV infected), both in vivo and / or when used in assays, including IHC, would be highly desirable.

[0038] Moreover, as noted above, the level of polymorphisms for US28 amongst clinical strains is higher than the level of polymorphisms previously reported for other HCMV- encoded proteins (Burrel et al, supra) and numerous N-terminal polymorphisms of US28 have been reported (Goffard et al., 2006, supra; Arav-Boger et al., 2002, supra).

[0039] The present inventor therefore considered the possibility that the presence of high levels of polymorphisms in the N-terminal region of US28, as bound by VUN100, may render the VUN 100 VHH molecule incapable of binding consistently to different HCMV strains. Indeed, when considered in that context, the results in relation to the binding of VUN100 to the different HCMV strains in Figure 8D of WO 2019 / 151865 show a difference in binding between the VHL / E, Merlin and TB40 / E strains of HCMV, with binding being particularly reduced in strain TB40 / E (Bl type) at around only half the level of binding observed against the Merlin strain. Moreover, further characterisation of VUN100 is reported in a pre-printed article available online by De Groof et a / , 2020 (doi: https: / / doi.org / 10.1101 / 2020.05.12.071860), wherein Fig 2 of the supplementary data gives the results of the % induced IE expression in the nucleus of CD14+ monocytes bound by VUN100. All cells tested were from HCMV seropositive individuals, and confirmed to be latently infected with HCMV, although the strain(s) of HCMV infecting each donor were undetermined. The level of IE expression induced by VUN100 binding to these HCMV-positive CD14+ cells from each of the four different patients varied substantially, with the reported figures being 57%, 33%, 22% and 4% (a range of difference of greater than 14-fold), respectively for cells from donors 1-4. This high level of response variability following the binding of VUN 100 to the confirmed HCMV-positive cells seems to be most likely due to the infection of each of the donors with different HCMV strains, and thus a strong indication that the binding ability of VUN 100 will vary considerably between different strains of HCMV. The same figure also shows high levels of response variability (in excess of 7-fold levels of difference) following the binding of a bivalent form of VUN 100 (termed VUN 100b by De Groof et al, 2020 (supra) as represented by SEQ ID NO: 63 of the present application) to the same group of HCMV-positive cells from the donors 1-4, again providing results indicative of strain-specific binding sensitivities.

[0040] In order to make use of US28 as a therapeutic target, it is important to overcome one or more of the most important obstacles, described above, not only including the relatively high levels of non-specific binding activity that has been reported for the art-known VUN 100 molecule, but also to minimise off-target binding activity, and to provide US28-binding molecules that overcome obstacles of strain diversity, viral mutations, mutagenic drift and the ability of the virus to hide from the immune system during viral latency.

[0041] It is therefore an object of the present invention to provide binding molecules which can bind highly specifically to biological materials that express US28 and / or which are positive for HCMV infection, compared to healthy human cells which should be much lower and / or to minimise the absolute levels of off-target binding to healthy human cells. For example, the provision of binding molecules that provide, or direct, a cytotoxic effect to the cells to which the binding molecules become bound are of great interest for combatting HCMV infections and conditions associated therewith, and it is an object of the invention to provide binding molecules that can target these effects in a way that minimises or avoids unacceptable (e.g. therapeutically-unacceptable) levels of off-target cytotoxic effects in healthy human cells. In particular, it is one of the objects of the present invention to provide binding molecules having a higher level of specificity for US28 and / or HCMV-infected cells than the VUN 100 Ab of WO 2019 / 151865 and De Groof et al, 2019 (supra) and / or than the VUN 100b bivalent molecule of De Groof et al, 2020 (supra), when assessed for specificity in binding to biological materials (e.g. cells) that express US28 and / or which are positive for HCMV infection compared to biological materials (e.g. equivalent cells) that do not express US28 and which are not positive for HCMV infection. It is another object of the present invention to provide binding molecules against US28 that are specific for the binding of biological materials that express US28 and / or which are positive for HCMV infected US28-expressing cells, compared to corresponding biological materials that do not express US28 (such as healthy human cells), and / or which display absolute levels of off-target binding to healthy human cells that are markedly lower than the VUN 100 Ab molecules as noted above.

[0042] It is a further object of the present invention to provide binding molecules which are strain agnostic, and ideally therefore capable of targeting all, or substantially all, HCMV infections irrespective of the strain, or combination of strains, of HCMV present and / or capable of providing an ongoing effect during the course of treatment of HCMV infections, despite the possibility of the rise of one or more HCMV mutations in the infecting strain(s) within the individual(s) being treated. In particular, binding molecules against US28 that have binding characteristics that show a greater degree of strain agnostic binding than the VUN 100 Ab are of particular interest.

[0043] SUMMARY OF THE INVENTION

[0044] The present invention provides binding molecules having one or more (preferably all) of highly specific binding to the US28 protein of human cytomegalovirus (HCMV), very low levels of non-specific binding to healthy (non-infected) cells, and / or a strain-agnostic binding ability, as well as nucleic acid molecules encoding the said binding molecules.

[0045] The binding molecules of the present invention are designed to bind within extracellular domain 3 (ECD3) of a US28 protein of human cytomegalovirus (HCMV).

[0046] The binding molecules of the present invention have been demonstrated to have excellent binding properties, including those described above, and as further described herein. For example, the binding molecules of the present invention have also surprisingly been demonstrated to provide particularly advantageous binding specificity for aggressive and / or metastasizing HCMV-infected cancers, including breast cancers.

[0047] In certain preferred embodiments, the binding molecule is selected from an antibody (including, for example, a BiTE antibody) and a chimeric antigen receptor (CAR), or functional variants, fragments, fusion proteins, and / or conjugates thereof, as well as nucleic acid molecules encoding the same. Said binding molecule may, for example, include or be bound to a cytotoxic component or other effector component, having the ability to exert an influence on (such as to inhibit or kill) any cells bound by the binding molecule. Said binding molecule may, for example, include or be bound to a component that can recruit other agents (e.g. other proteins, drugs, cells or any other substance of choice) in such a way that the recruited agent has a specifically- targeted ability to exert an influence on (such as to inhibit or kill) cells bound by the binding molecule; a non-limiting example therefore is a BiTE molecule, which possesses the ability to recruit a T-cell to act upon cells bound by said BiTE. Also provided are cells expressing said binding molecules, including examples in which the binding molecule is a CAR, and said cells may be CAR-expressing cells, including CAR- T cells, CAR-NK cells, and CAR-M cells.

[0048] These, and further disclosures of the present invention are described in more detail by the following description and the appended claims and figures.

[0049] Accordingly, a first aspect of the present invention provides binding molecule, comprising one or more polypeptide chains, said binding molecule having binding specificity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of human cytomegalovirus (HCMV). In accordance with the present invention, ECD3 of the US28 protein comprises, consists essentially of, or consist of, an amino acid sequence presented in the US28 protein encoded by a strain of HCMV at positions corresponding to positions 167 to 183 of the US28 protein encoded by the DB strain of human cytomegalovirus (HCMV) as set forth in SEQ ID NO: 5.

[0050] For example, and without limitation, a binding molecule of the first aspect of the present invention may be selected from an antibody or a chimeric antigen receptor (CAR).

[0051] The binding molecule of the first aspect of the present invention may, for example, have binding specificity to an epitope present entirely within extracellular domain 3 (ECD3) of the US28 protein of HCMV.

[0052] The binding molecule of the first aspect of the present invention may, for example, have binding specificity to a linear epitope within ECD3 of the US28 protein.

[0053] The binding molecule of the first aspect of the present invention may, for example, have a strain agnostic binding specificity to an epitope within ECD3 of a US28 protein of HCMV. For example, the binding molecule may have a binding specificity to an epitope within ECD3 of a US28 protein of HCMV, wherein the binding specificity is agnostic to two or more (such as all) of HCMV strains, for example agnostic to 4D- variant strains and 4N-variant strains (each as described further herein), optionally two or more (such as all) HCMV strains selected from the group consisting of DB, Towne, AF1, VHL / E, AD169, BL, DAVIS, JP, Merlin, PH, TB40 / E, Toledo, TR and VR1814 (FIX).

[0054] Accordingly, the binding molecule of the first aspect of the present invention may, for example, have binding specificity to an epitope within ECD3 of the US28 protein of HCMV, irrespective of whether the ECD3 of the US28 protein comprises the sequence of a 4D-variant or a 4N-variant: - wherein the 4D-variant comprises the sequence of TKKDNQCMTDYDYLEVS (SEQ ID NO: 7) as found in ECD3 of US28 as encoded by a first group of HCMV strains, such as Towne, VR1814, TB40 / E, Merlin, JP, Ad 169, AF1, VHL / E, BL and DAVIS; and

[0055] - wherein the 4N-variant comprises the sequence of TKKNNQCMTDYDYLEVS (SEQ ID NO: 6) as found in ECD3 of US28 as encoded by a second group of HCMV strains, such as Toledo, TR and DB strains.

[0056] By making use of the protocols described herein, the applicant has consistently and repeatedly generated numerous anti-ECD3 antibodies with one or more of the above-noted beneficial binding properties, including antibodies referred to herein by the following designations (the sequences of which are also provided below): US28- 13-5G6-1D3; US28-13-1C10-1C10; US28-13-1H3-1A10; US28-14-2C2-1G4; US28- 13-1C10-1G9; and US28-14-4E4-1E8.

[0057] In one embodiment, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 13-5G6-1D3 (generally abbreviated herein to "1D3") as defined by SEQ ID NOs: 8, 9, 10, 14, 15 and 16, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1D3, and optionally wherein the binding molecule is selected from an antibody and a CAR.

[0058] Optionally, the binding molecule according to this embodiment may comprise (a) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable heavy chain (VH) of antibody 1D3, as defined by SEQ ID NOs: 8, 9, and 10, respectively; and / or (b) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable light chain (VL) of antibody 1D3, as defined by SEQ ID NOs: 14, 15, and 16, respectively.

[0059] Additionally, or alternatively, in a further option the binding molecule according to this embodiment may comprise: (a) at least one variable heavy chain (VH) polypeptide that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID Nos: 8, 9, and 10, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 12; and / or (b) at least one variable light chain (VL) polypeptide that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 14, 15, and 16, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 18. The following sequences of the US28-13-5G6-1D3 antibody are identified herein, with reference to the sequence identification numbers (SEQ ID NOs) described below:

[0060] In another embodiment, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six CDRs corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 13-1C10-1C10 (generally abbreviated herein to "1C10") as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1C10, and optionally wherein the binding molecule is selected from an antibody and a CAR.

[0061] Optionally, the binding molecule according to this embodiment may comprise (a) one, two, or all three, of the CDR 1, 2, and 3, sequences of the VH of antibody 13- 1C10-1C10, as defined by SEQ ID NOs: 112, 113, and 114, respectively; and / or (b) one, two, or all three, of the CDR 1, 2, and 3, sequences of the VL of antibody 13- 1C10-1C10, as defined by SEQ ID NOs: 117, 83, and 118, respectively.

[0062] Additionally, or alternatively, in a further option the binding molecule according to this embodiment may comprise: (a) at least one VH polypeptide that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID NOs: 112, 113, and 114, respectively, and optionally wherein the at least one VH polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 104; and / or (b) at least one VL polypeptide that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 117, 83, and 118, respectively, and optionally wherein the VL polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 108. The following sequences of the US28-13-1C10-1C10 antibody are identified herein, with reference to the sequence identification numbers (SEQ ID NOs) described below:

[0063] In another embodiment, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six CDRs corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 13-1H3-1A10 (generally abbreviated herein to "1A10") as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1A10, and optionally wherein the binding molecule is selected from an antibody and a CAR.

[0064] Optionally, the binding molecule according to this embodiment may comprise (a) one, two, or all three, of the CDR 1, 2, and 3, sequences of the VH of antibody 13- 1H3-1A10, as defined by SEQ ID NOs: 112, 113, and 114, respectively; and / or (b) one, two, or all three, of the CDR 1, 2, and 3, sequences of the VL of antibody 13-1H3- 1A10, as defined by SEQ ID NOs: 117, 83, and 118, respectively.

[0065] Additionally, or alternatively, in a further option the binding molecule according to this embodiment may comprise: (a) at least one VH polypeptide that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID NOs: 112, 113, and 114, respectively, and optionally wherein the at least one VH polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 122; and / or (b) at least one VL polypeptide that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 117, 83, and 118, respectively, and optionally wherein the VL polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 126. The following sequences of the US28-13-1H3-1A10 antibody are identified herein, with reference to the sequence identification numbers (SEQ ID NOs) described below:

[0066] In another embodiment, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six CDRs corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 13-1C10-1G9 (generally abbreviated herein to "1G9") as defined by SEQ ID NOs: 76, 77, 78, 82, 83 and 84, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1G9, and optionally wherein the binding molecule is selected from an antibody and a CAR.

[0067] Optionally, the binding molecule according to this embodiment may comprise (a) one, two, or all three, of the CDR 1, 2, and 3, sequences of the VH of antibody 13- 1C10-1G9, as defined by SEQ ID NOs: 76, 77, and 78, respectively; and / or (b) one, two, or all three, of the CDR 1, 2, and 3, sequences of the VL of antibody 13-1C10- 1G9, as defined by SEQ ID NOs: 82, 83, and 84, respectively.

[0068] Additionally, or alternatively, in a further option the binding molecule according to this embodiment may comprise: (a) at least one VH polypeptide that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID NOs: 76, 77, and 78, respectively, and optionally wherein the at least one VH polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 68; and / or (b) at least one VL polypeptide that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 82, 83, and 84, respectively, and optionally wherein the VL polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 72. The following sequences of the US28-13-1C10-1G9 antibody are identified herein, with reference to the sequence identification numbers (SEQ ID NOs) described below:

[0069] In another embodiment, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six CDRs corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 14-4E4-1E8 (generally abbreviated herein to "1E8") as defined by SEQ ID NOs: 76, 95, 96, 82, 99 and 100, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1E8, and optionally wherein the binding molecule is selected from an antibody and a CAR.

[0070] Optionally, the binding molecule according to this embodiment may comprise (a) one, two, or all three, of the CDR 1, 2, and 3, sequences of the VH of antibody 14- 4E4-1E8, as defined by SEQ ID NOs: 76, 95, and 96, respectively; and / or (b) one, two, or all three, of the CDR 1, 2, and 3, sequences of the VL of antibody 14-4E4-1E8, as defined by SEQ ID NOs: 82, 99, and 100, respectively.

[0071] Additionally, or alternatively, in a further option the binding molecule according to this embodiment may comprise: (a) at least one VH polypeptide that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID NOs: 76, 95, and 96, respectively, and optionally wherein the at least one VH polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 88; and / or (b) at least one VL polypeptide that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 82, 99, and 100, respectively, and optionally wherein the VL polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 92. The following sequences of the US28-14-4E4-1E8 antibody are identified herein, with reference to the sequence identification numbers (SEQ ID NOs) described below:

[0072] In another embodiment, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six CDRs corresponding to any one, two, three, four, five or all six of the following consensus CDR sequences (wherein replacement amino acids for a particular position are indicated in parenthesis, and * indicates an absence of an amino acid at that position):

[0073] (a) VH-CDR1 corresponding to S(Y / H)A(M / L)S (SEQ ID NO: 167);

[0074] (b) VH-CDR2 corresponding to SISS(G / R)G(S / R)TYYPDSVKG (SEQ ID NO: 168);

[0075] (c) VH-CDR3 corresponding to GG(S / T)(T / R / H)(M / H / Y)(I / S)(T / Y) (T / G)(G / N)(L / *)GF(A / D)(Y / F) (SEQ ID NO: 169);

[0076] (d) VL-CDRI corresponding to S(A / V)SSSVSYMH (SEQ ID NO: 170);

[0077] (e) VL-CDR2 corresponding to D(T / S)SKLAS (SEQ ID NO: 171); and / or

[0078] (f) VL-CDR3 corresponding to QQW(S / T / *)SN(* / N)PP(I / L)T (SEQ ID NO: 172).

[0079] In another embodiment, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six CDRs corresponding to any one, two, three, four, five or all six of the following consensus CDR sequences (wherein replacement amino acids for a particular position are indicated in parenthesis, and * indicates an absence of an amino acid at that position):

[0080] (a) VH-CDR1 corresponding to S(Y / H)A(M / L)S (SEQ ID NO: 167);

[0081] (b) VH-CDR2 corresponding to SISS(G / R)GRTYYPDSVKG (SEQ ID NO: 174); (c) VH-CDR3 corresponding to GG(S / T)(T / R / H)(M / H / Y)(I / S)(T / Y) (T / G)(G / N)GF(A / D)(Y / F) (SEQ ID NO: 175);

[0082] (d) VL-CDR1 corresponding to S(A / V)SSSVSYMH (SEQ ID NO: 170);

[0083] (e) VL-CDR2 corresponding to D(T / S)SKI_AS (SEQ ID NO: 171); and / or

[0084] (f) VL-CDR3 corresponding to QQW(T / *)SN(* / N)PPIT (SEQ ID NO: 176).

[0085] Functional variants of any one or more of the VH-CDR1, VH-CDR2, VH-CDR3, VL- CDR1, VL-CDR2 and / or VL-CDR3 sequences as defined above for any of antibodies US28-13-5G6-1D3, 13-1C10-1C10, 13-1H3-1A10, 13-1C10-1G9, 14-4E4-1E8 may optionally comprise one of the consensus sequences as set forth above for the VH- CDR1, VH-CDR2, VH-CDR3, VL-CDR1, VL-CDR2 and / or VL-CDR3 sequences, respectively.

[0086] Sequences corresponding to the above-noted SEQ ID NOs are given in the section of this application entitled "Sequences", which follows the Examples, in this application.

[0087] In certain embodiments, a binding molecule according to the first aspect of the present invention may be selected from the group consisting of:

[0088] (a) bivalent antibodies, such as IgG-scFv antibodies (for example, wherein a first binding domain is an intact IgG and a second binding domain is an scFv attached to the first binding domain at the N-terminus of a light chain and / or at the C-terminus of a light chain and / or at the N-terminus of a heavy chain and / or at the C-terminus of a heavy chain of the IgG, or vice versa),'

[0089] (b) monovalent antibodies, such as a DuoBody® or 'knob-in-hole' bispecific antibody (for example, an scFv-KIH, scFv-KIHr, a BiTE-KIH or a BiTE-KIHr;

[0090] (c) scFvz-Fc antibodies;

[0091] (d) bispecific antibodies, such as bispecific T-cell engager (BiTE) antibodies;

[0092] (e) dual variable domain (DVD)-Ig antibodies;

[0093] (f) dual-affinity re-targeting (DART)-based antibodies (for example, DART2-

[0094] Fc or DART);

[0095] (g) trispecific antibodies, such as DNL-Fabs antibodies;

[0096] (h) scFv-HSA-scFv antibodies;

[0097] (I) single domain antibodies;

[0098] (j) heavy-chain-only IgGs (hdgGs), such as camelid IgG (e.g. VHH antibodies) and shark immunoglobulin new antigen receptor (IgNAR), and single chain antibodies thereof; and

[0099] (k) a chimeric antigen receptor (CAR) comprising an extracellular domain that composes, consists essentially of, or consists of a binding molecule according to the first aspect of the present invention, for example an extracellular domain that comprises any one of options (a) to (h) of this list, or combinations thereof. Additionally, or alternatively, a binding molecule according to the first aspect of the present invention may be selected from the group consisting of:

[0100] (i) a bispecific immune cell engager antibody, for example, a bispecific T- cell engager (BiTE), and optionally wherein the BiTE antibody comprises a CD3-binding domain; or

[0101] (ii) monoclonal antibody, optionally a recombinant monoclonal antibody, for example, a monoclonal antibody produced recombinantly by CHO cells.

[0102] The first aspect of the present invention also provides a functional fragment of a binding molecule as defined above, wherein the functional fragment:

[0103] (a) comprises or consists of an antigen-binding fragment of a binding molecule as defined by any of the preceding claims, or a variant, fusion or derivative thereof selected from the group consisting of: an Fv fragment (such as a single chain Fv fragment (scFv), or a disulphide-bonded Fv fragment), a Fab-like fragment (such as a Fab fragment, a Fab' fragment or a F(ab)2 fragment), and single domain antibodies (dAbs, including single and dual formats, such as dAb-linker-dAb and nanobodies);

[0104] (b) provides one or more of the binding characteristics of a binding molecule of the first aspect of the present invention, as defined herein;

[0105] (c) comprises the CDR sequences of a binding molecule of the first aspect of the present invention, as defined herein; and / or

[0106] (d) comprises the VH and / or VL sequences of a binding molecule of the first aspect of the present invention, as defined herein.

[0107] Fusions of the binding molecules of the first aspect of the present invention are also provided herein. For example, a binding molecule as defined above, or a functional fragment of said binding molecule as defined above, is provided wherein the binding molecule or the functional fragment thereof comprises a fusion polypeptide sequence, said fusion polypeptide sequence comprising a first amino acid sequence fused to a second amino acid sequence, wherein the first amino acid sequence comprises or consists of at least one of the polypeptide chains of the binding molecule or of the functional fragment thereof, and the second amino acid sequence is a fusion partner.

[0108] In certain embodiments, the binding molecule of the first aspect of the present invention is, or comprised within, a chimeric antigen receptor (CAR). Accordingly, the first aspect of the present invention also provides a CAR comprising:

[0109] (i) an extracellular domain, wherein the extracellular domain comprises or consists of a binding molecule of the first aspect of the present invention as defined herein, or a functional fragment of said binding molecule as defined herein;

[0110] (ii) a transmembrane domain; and

[0111] (iii) an intracellular domain; wherein the extracellular domain of the CAR has binding specificity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of human cytomegalovirus (HCMV), and wherein ECD3 of the US28 protein comprises an amino acid sequence presented in the US28 protein at positions corresponding to positions 167 to 183 of the US28 protein encoded by human cytomegalovirus (HCMV) as set forth in SEQ ID NO: 5

[0112] Optionally, in said CAR:

[0113] (a) the extracellular domain of the CAR has binding specificity to an epitope present entirely within extracellular domain 3 (ECD3) of the US28 protein of HCMV;

[0114] (b) the extracellular domain of the CAR has binding specificity to a linear epitope within ECD3 of the US28 protein;

[0115] (c) the extracellular domain of the CAR has binding specificity to an epitope within ECD3 of a US28 protein of HCMV that is HCMV strain agnostic, for example, binding specificity to an epitope within ECD3 of a US28 protein of HCMV that is agnostic to two or more (such as all) of HCMV strains selected from the group consisting of DB, Towne, AD169, DAVIS, BL, JP, Merlin, PH, TB40 / E, Toledo, TR, VHL / E and VR1814 (FIX); and / or

[0116] (d) the extracellular domain of the CAR has specificity to an epitope within ECD3 of the US28 protein of HCMV, irrespective of whether the ECD3 of the US28 protein comprises the sequence of:

[0117] - TKKDNQCMTDYDYLEVS (SEQ ID NO: 7) as found in ECD3 of US28 as encoded by a majority of HCMV strains, or

[0118] - TKKNNQCMTDYDYLEVS (SEQ ID NO: 6) as found in ECD3 of US28 as encoded by a minority of HCMV strains.

[0119] In certain embodiments, in a CAR according to the first aspect of the present invention:

[0120] (a) the extracellular domain of the CAR comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 1D3 as defined by SEQ ID NOs: 8, 9, 10, 14, 15 and 16, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1D3;

[0121] (b) the extracellular domain of the CAR comprises:

[0122] (i) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable heavy chain (VH) of antibody 1D3, as defined by SEQ ID NOs: 8, 9, and 10, respectively; and / or

[0123] (ii) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable light chain (VL) of antibody 1D3, as defined by SEQ ID NOs: 14, 15, and 16, respectively; and / or (c) the extracellular domain of the CAR comprises: (i) at least one variable heavy chain (VH) polypeptide sequence that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID Nos: 8, 9, and 10, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 12; and / or (ii) at least one variable light chain (VL) polypeptide sequence that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 14, 15, and 16, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 18.

[0124] In another embodiment, in a CAR according to the first aspect of the present invention:

[0125] (a) the extracellular domain of the CAR comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 1C10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1C10;

[0126] (b) the extracellular domain of the CAR comprises:

[0127] (i) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable heavy chain (VH) of antibody 1C10, as defined by SEQ ID NOs: 112, 113, and 114, respectively; and / or

[0128] (ii) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable light chain (VL) of antibody 1C10, as defined by SEQ ID NOs: 117, 83, and 118, respectively; and / or

[0129] (c) the extracellular domain of the CAR comprises: (i) at least one variable heavy chain (VH) polypeptide sequence that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID Nos: 112, 113, and 114, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 104; and / or (ii) at least one variable light chain (VL) polypeptide sequence that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 117, 83, and 118, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 108.

[0130] In another embodiment, in a CAR according to the first aspect of the present invention:

[0131] (a) the extracellular domain of the CAR comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 1A10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1A10;

[0132] (b) the extracellular domain of the CAR comprises:

[0133] (i) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable heavy chain (VH) of antibody 1A10, as defined by SEQ ID NOs: 112, 113, and 114, respectively; and / or

[0134] (ii) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable light chain (VL) of antibody 1A10, as defined by SEQ ID NOs: 117, 83, and 118, respectively; and / or

[0135] (c) the extracellular domain of the CAR comprises: (i) at least one variable heavy chain (VH) polypeptide sequence that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID Nos: 112, 113, and 114, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 122; and / or (ii) at least one variable light chain (VL) polypeptide sequence that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 117, 83, and 118, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 126.

[0136] In another embodiment, in a CAR according to the first aspect of the present invention:

[0137] (a) the extracellular domain of the CAR comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 1G9 as defined by SEQ ID NOs: 76, 77, 78, 82, 83 and 84, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1G9;

[0138] (b) the extracellular domain of the CAR comprises:

[0139] (i) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable heavy chain (VH) of antibody 1G9, as defined by SEQ ID NOs: 76, 77, and 78, respectively; and / or

[0140] (ii) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable light chain (VL) of antibody 1G9, as defined by SEQ ID NOs: 82, 83, and 84, respectively; and / or

[0141] (c) the extracellular domain of the CAR comprises: (i) at least one variable heavy chain (VH) polypeptide sequence that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID Nos: 76, 77, and 78, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 68; and / or (ii) at least one variable light chain (VL) polypeptide sequence that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 82, 83, and 84, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 72.

[0142] In another embodiment, in a CAR according to the first aspect of the present invention:

[0143] (a) the extracellular domain of the CAR comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 1E8 as defined by SEQ ID NOs: 76, 95, 96, 82, 99 and 100, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1E8;

[0144] (b) the extracellular domain of the CAR comprises:

[0145] (i) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable heavy chain (VH) of antibody 1E8, as defined by SEQ ID NOs: 76, 95, and 96, respectively; and / or

[0146] (ii) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable light chain (VL) of antibody 1E8, as defined by SEQ ID NOs: 82, 99, and 100, respectively; and / or

[0147] (c) the extracellular domain of the CAR comprises: (I) at least one variable heavy chain (VH) polypeptide sequence that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID Nos: 76, 95, and 96, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 88; and / or (ii) at least one variable light chain (VL) polypeptide sequence that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 82, 99, and 100, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 92.

[0148] Optionally, in said CAR, the extracellular domain is an antibody, for example a single-chain variable fragment (scFv).

[0149] The transmembrane domain of a CAR according to the first aspect of the present invention may, for example, comprise the transmembrane domain of a protein, for example the transmembrane domain of a transmembrane receptor protein, and optionally wherein the transmembrane domain comprises the transmembrane domain of a protein selected from the group consisting of the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD8, CD45 and CD4.

[0150] The extracellular domain of a CAR according to the first aspect of the present invention may, for example, be connected to the transmembrane domain by a hinge region. The intracellular domain of a CAR according to the first aspect of the present invention may, for example, comprise an intracellular signalling domain, for example wherein: (a) the intracellular signalling domain comprises one or more immunoreceptor tyrosine-based activation motifs (ITAMs); and / or (b) the intracellular signalling domain comprises a signalling domain of CD3 zeta, Fc receptor gamma, Fc receptor beta, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, and CD66d.

[0151] The intracellular domain of a CAR according to the first aspect of the present invention may, for example, comprise one or more costimulatory domains, for example: (a) wherein the one or more costimulatory domains includes one or more functional signalling domains obtained from a protein selected from the group consisting of CD28, 41BB, 0X40, ICOS, CD27, and DAP10; (b) wherein the intracellular domain incorporates a costimulatory domain proximal to the intracellular signalling domain, (c) wherein the intracellular domain comprises two or more costimulatory domains, for example two in-line costimulatory domains, and / or (d) wherein the intracellular domain incorporates separate cytokine signals.

[0152] A CAR according to the first aspect of the present invention may, for example, additionally comprise a leader sequence.

[0153] A second aspect of the present invention provides a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, wherein the nucleic acid molecule comprises, or the combination of multiple distinct nucleic acid molecules collectively comprise, one or more nucleic acid sequences that, individually or in combination, encode the binding molecule of the first aspect of the present invention, for example an antibody or CAR according to the first aspect of the present invention.

[0154] A third aspect of the present invention provides a vector comprising (or combination of multiple distinct vectors which collectively comprise) a nucleic acid molecule according to the second aspect of the present invention, or combination of multiple distinct nucleic acid molecules according to the second aspect of the present invention. The, or each, vector of the third aspect of the present invention may, for example, be selected from the group consisting of a retroviral vector, a plasmid, a lentivirus vector, and an adenoviral vector.

[0155] Optionally a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules according to the second aspect of the present invention, and / or a vector according to the third aspect of the present invention, comprise one or more additional sequences, wherein the or each additional sequence encodes one or more selectable markers.

[0156] A fourth aspect of the present invention provides a cell, or a population of cells (optionally a homogeneous or heterogeneous population of cells) comprising the nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the second aspect of the present invention, and / or a vector according to the third aspect of the present invention, optionally wherein the cell expresses one or more binding molecules according to the first aspect of the present invention (such as one or more antibodies, and / or one or more CARs), said one or more binding molecules and / or CARs being encoded by the nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the second aspect of the present invention, or a vector according to the third aspect of the present invention. Said cells may, optionally, be selected from isolated cells, ex vivo cells, and in vitro cells.

[0157] A cell according to the fourth aspect of the present invention may, for example, comprise: (a) a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the second aspect of the present invention, wherein the encoded binding molecule is an antibody, a functional fragment of said antibody, or an antibody of functional fragment thereof that comprises a fusion polypeptide sequence, according to the first aspect of the present invention; and / or (b) a vector according to the third aspect of the present invention, wherein said vector comprises a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules as defined by option (a) of this paragraph.

[0158] A cell according to the fourth aspect of the present invention may, for example, comprise: (a) a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the second aspect of the present invention, wherein the encoded binding molecule is a CAR according to the first aspect of the present invention; and / or (b) a vector according to the third aspect of the present invention, wherein said vector comprises a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules as defined by part (a) of this paragraph. Without limitation, said cell may, for example, be selected from the group consisting of: a T cell, natural killer (NK) cell, and a macrophage. Accordingly, the cell may optionally be a CAR-T cell, a CAR-NK cell or a CAR-macrophage, and optionally, when the cell is a CAR-T cell, then for example the T-cell may be selected from the group consisting of CD8+T cells, CD4+ T cells, effector T cells, helper T cells, memory T cells, cytotoxic T lymphocytes (CTLs) , EBV-specific T cell receptor (TCR) or y6-T cell subtypes.

[0159] The fourth aspect of the present invention also provides a cell comprising a binding molecule according the first aspect of the present invention and / or a nucleic acid encoding said binding molecule, optionally wherein said nucleic acid is a nucleic acid or vector as defined by the second or third aspects of the present invention, respectively. For example, the binding molecule may be an antibody according the first aspect of the present invention, a functional fragment of said antibody according the first aspect of the present invention, or an antibody of functional fragment thereof that comprises a fusion polypeptide sequence according the first aspect of the present invention, and optionally wherein the antibody is monoclonal antibody, and further for example wherein the cell is a mammalian cell, such as a CHO cell, that recombinantly expresses the monoclonal antibody.

[0160] The fourth aspect of the present invention also provides a cell comprising a CAR according to the first aspect of the present invention and / or a nucleic acid encoding said CAR, optionally wherein said nucleic acid is a nucleic acid or vector as defined by the second or third aspects of the present invention, respectively. Without limitation, said cell may, for example, be selected from the group consisting of: a T cell, natural killer (NK) cell, and a macrophage. Accordingly, the cell may optionally be a CAR-T cell, a CAR-NK cell or a CAR-macrophage, and optionally, when the cell is a CAR-T cell, then for example the T-cell may be selected from the group consisting of CD8+T cells, CD4+ T cells, effector T cells, helper T cells, memory T cells, cytotoxic T lymphocytes (CTLs) , EBV-specific T cell receptor (TCR) or y6-T cell subtypes.

[0161] A fifth aspect of the present invention provides a method of producing a cell, more particularly a recombinant cell, or a population of such cells (optionally a homogeneous or heterogeneous population of cells), the method comprising introducing a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the second aspect of the present invention, and / or a vector according to third aspect of the present invention, into a cell.

[0162] Said method optionally further comprises a step of selecting cells according to the fifth aspect of the present invention; for example selecting said cells from a heterogeneous cell population, thereby to create an enriched and / or homogeneous cell population. Said selection step may include selecting for the presence of one or more selectable markers present in the nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the second aspect of the present invention, and / or a vector according to third aspect of the present invention.

[0163] A sixth aspect of the present invention provides a method of producing a binding molecule according to the first aspect of the present invention, for example an antibody or a CAR according to the first aspect of the present invention, the method comprising: expressing a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the second aspect of the present invention, and / or a vector according to the third aspect of the present invention, in a cell, more particularly a recombinant cell, or a population of such cells (optionally a homogeneous or heterogeneous population of cells). Optionally, the method of the sixth aspect of the present invention may also comprise the step of isolating the thus-produced binding molecule from the cell; for example, wherein the binding molecule is an antibody according to the first aspect of the present invention, a functional fragment of said antibody, or an antibody of functional fragment thereof that comprises a fusion polypeptide sequence according to the first aspect of the present invention. Said cells may, optionally, be selected from isolated cells, ex vivo cells, and in vitro cells.

[0164] A seventh aspect of the present invention provides an isolated binding molecule that is obtained, or obtainable, by the method of the sixth aspect of the present invention, optionally, wherein the isolated binding molecule is further formulated for administration to a subject.

[0165] An eighth aspect of the present invention provides a conjugate, the conjugate comprising a moiety conjugated to a binding molecule as defined by the first aspect of the present invention, or to a functional fragment of said binding molecule. Without limitation, said moiety may for example be a therapeutic, prophylactic, diagnostic, prognostic, or theragnostic moiety. In some embodiments, the moiety is a drug (for example, wherein the conjugate is an antibody-drug conjugate ("ADC")) and / or a radioactive moiety (for example, wherein the conjugate is suitable for use in radioimmunotherapy ("RIT")).

[0166] A ninth aspect of the present invention provides a method of producing a conjugate according to the eighth aspect of the present invention, the method comprising the steps of:

[0167] (a) providing a binding molecule as defined by the first aspect of the present invention, or a functional fragment of said binding molecule; and

[0168] (b) conjugating a moiety to the binding molecule, or to the functional fragment of said binding molecule.

[0169] The method of the ninth aspect of the present invention may additionally comprise the step of isolating the thus-produced conjugate. The isolated conjugate may therefore be presented in an isolated form, for example in the form of a composition wherein at least 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or substantially 100% (by molar ratio) of the binding molecule, or the functional fragment of said binding molecule, is present in the form of the conjugate. Additionally or alternatively, the isolated form of the conjugate may be a composition wherein at least 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or substantially 100% (by molar ratio) of the moiety, is present in the form of the conjugate. The ninth aspect of the present invention further provides the isolated conjugate, or a composition comprising said isolated conjugate.

[0170] In certain preferred embodiments of the conjugate of the eighth aspect of the present invention, or of the method of the ninth aspect of the present invention, the binding molecule is an antibody according to the first aspect of the present invention, a functional fragment of said antibody, or an antibody of functional fragment thereof that comprises a fusion polypeptide sequence according to the first aspect of the present invention.

[0171] A tenth aspect of the present invention provides an isolated conjugate that is obtained, or obtainable, by the method of the ninth aspect of the present invention, optionally, wherein the isolated conjugate is further formulated for administration to a subject.

[0172] An eleventh aspect of the present invention provides a method of combating HCMV or a disease or condition associated with HCMV, the method comprising administering to a subject, or to ex vivo or in vitro cellular material, any one or more agents selected from the group consisting of: i. a binding molecule according to the first aspect of the present invention,

[0173] II. a functional fragment of said binding molecule according to the first aspect of the present invention, ill. an isolated binding molecule according to the seventh aspect of the present invention, iv. a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the second aspect of the present invention, v. a vector according to the third aspect of the present invention, vi. a cell according to the fourth aspect of the present invention, vii. a conjugate according to the eighth aspect of the present invention, and viii. an isolated conjugate according to the tenth aspect of the present invention.

[0174] To put it another way, the eleventh aspect of the present invention provides one or more of said agents for use in combating a disease or condition associated with HCMV in a subject, or in ex vivo or in vitro cellular material.

[0175] Further, the eleventh aspect of the present invention provides for the use one or more of said agents in the manufacture of a medicament for combating a disease or condition associated with HCMV in a subject, or in ex vivo or in vitro cellular material.

[0176] The disease or condition associated with HCMV, to be combatted in accordance with the eleventh aspect of the present invention may be an HCMV infection or be associated with an HCMV infection. The HCMV infection may, in one embodiment, be a single strain infection. Optionally, in an alternative embodiment, the HCMV infection comprises a multi-strain HCMV infection, wherein the multi-strain HCMV infection comprises infection with more than one different strain of HCMV, for example two or more HCMV strains that encode different US28 protein sequences. Said two or more strains may encode US28 proteins that differ in one or more of the extracellular regions, such as in the N-terminal (ECD1) regional, the first extracellular loop (ECD2) region, the second extracellular loop (ECD3) region, and / or the third extracellular loop (ECD4) region. In one embodiment of interest, the two or more HCMV strains in a multi-strain HCMV infection each encode a US28 protein that differs from the other at least in one or more positions of the N-terminal (ECD1) region; for example they may differ at 1, 2, 3, 4, 5, 6, 8, 9, 10 or more amino acid positions in the N-terminal (ECD1) region. Additionally, or alternatively, in another embodiment of interest, the two or more HCMV strains in a multi-strain HCMV infection each encode a US28 protein that differs from the other at one or more positions of the second extracellular loop (ECD3) region, for example one or more of the HCMV strains in a multi-strain HCMV infection may encode a US28 protein that encodes the 4N-variant of ECD3, and one or more of the other HCMV strains in a multi-strain HCMV infection may encode a US28 protein that encodes the 4D-variant of ECD3.

[0177] The disease or condition associated with HCMV, to be combatted in accordance with the eleventh aspect of the present invention may be a latent HCMV infection (for example, a single or multi-strain latent HCMV infection) or be associated with a latent HCMV infection (optionally a multi-strain latent HCMV infection).

[0178] The disease or condition associated with HCMV, to be combatted in accordance with the eleventh aspect of the present invention may be a lytic HCMV infection (optionally a multi-strain lytic HCMV infection) or be associated with a lytic HCMV infection (optionally a multi-strain lytic HCMV infection).

[0179] The disease or condition associated with HCMV, to be combatted in accordance with the eleventh aspect of the present invention may be a congenital HCMV infection (for example, a single or multi-strain infection), such as a latent congenital single or multi-strain HCMV infection or a lytic congenital single or multi-strain HCMV infection;

[0180] The disease or condition associated with HCMV, to be combatted in accordance with the eleventh aspect of the present invention may be cancer, for example HCMV- infected cancer (optionally a single-strain, or multi-strain, HCMV infected cancer), such as latent HCMV-infected cancer (optionally a single-strain, or multi-strain, latent HCMV infected cancer).

[0181] The disease or condition associated with HCMV, to be combatted in accordance with the eleventh aspect of the present invention may be an epithelial cancer; optionally wherein the epithelial cancer is breast cancer; for example, wherein the breast cancer is triple negative breast cancer (TNBC), or a HER2-positive breast cancer. A HER2-positive breast cancer may, for example, be HER2+ HR- (wherein HR- means hormone receptor-negative, and refers to oestrogen receptor negative (ER-) and progesterone receptor negative (PR-) status); or HER2+ ER+ PR-; or HER2+ ER- PR+; or a triple positive form of breast cancer "TPBC" that is HER2+ ER+ PR+. It is noted that HER2+ HR- is a particularly aggressive form of breast cancer and is of high interest for diagnosis treatment in accordance with the present invention. Said forms of epithelial cancer may optionally be a single-strain, or multi-strain, form of HCMV infected epithelial cancer, for example a latent HCMV- infected form of epithelial cancer (optionally a single-strain, or multi-strain, latent HCMV infected cancer).

[0182] The disease or condition associated with HCMV, to be combatted in accordance with the eleventh aspect of the present invention, may be a metastasising and / or aggressive form of cancer. Said forms of metastasising and / or aggressive cancer may optionally be a single-strain, or multi-strain, form of HCMV infected metastasising and / or aggressive cancer, for example a latent HCMV-infected form of metastasising and / or aggressive cancer (optionally a single-strain, or multi-strain, latent HCMV infected cancer).

[0183] In some embodiments, the disease or condition associated with HCMV, to be combatted in accordance with the eleventh aspect of the present invention may be glioblastoma. In other embodiment described herein, the disease or condition is not glioblastoma and / or the subject to be treated does not have and / or has not been diagnosed as having glioblastoma.

[0184] In some embodiments, the subject (or the ex vivo or in vitro cellular material) to be treated in accordance with the eleventh aspect of the present invention may have, and / or have been diagnosed has having or possessing, HCMV-infected cancer cells, such as latent HCMV-infected cancer cells.

[0185] In some embodiments, the subject to be treated in accordance with the eleventh aspect of the present invention may be, or intended to be, the recipient of a cellular material, such as the donation of a cellular product. Said cellular product may, for example, comprise, consist essentially of, or consist of, living ex vivo cellular material selected from the group that includes: one or more types of ex vivo cells; one or more types of ex vivo cell cultures; one or more types of ex vivo tissues; one or more types of ex vivo tissue cultures; one or more types of ex vivo organs; and / or one or more types of ex vivo organ cultures. Optionally, the cellular product may be derived, directly or indirectly, from a living donor.

[0186] In some embodiments, the subject to be treated in accordance with the eleventh aspect of the present invention may be, or intended to be, the donor of a cellular material, such as the donor of a cellular product. Said cellular product may be comprise, consist essentially of, or consist of any one or more of cells, tissue or an organ from said donor.

[0187] In some embodiments, the ex vivo or in vitro cellular material to be treated in accordance with the eleventh aspect of the present invention may be an ex vivo cellular product. Said ex vivo cellular product may, for example, comprise, consist essentially of, or consist of, living ex vivo cellular material selected from the group that includes: one or more types of ex vivo cells; one or more types of ex vivo cell cultures; one or more types of ex vivo tissues; one or more types of ex vivo tissue cultures; one or more types of ex vivo organs; and / or one or more types of ex vivo organ cultures. Optionally, the cellular product may be derived, directly or indirectly, from a living donor.

[0188] In one embodiment, the one or more agents to be used in accordance with the eleventh aspect of the present invention may, for example, be (or include one or more agents) selected from the group consisting of:

[0189] I. a therapeutic antibody as defined by the first aspect of the present invention,

[0190] II. a functional fragment of said therapeutic antibody as defined by the first aspect of the present invention, ill. a therapeutic antibody that comprises a fusion polypeptide sequence as defined by the first aspect of the present invention; and iv. a functional fragment of said therapeutic antibody that comprises a fusion polypeptide sequence as defined by the first aspect of the present invention.

[0191] For example, the one or more agents used in accordance with the eleventh aspect of the present invention may be (or include one or more agents selected from) a bispecific antibody as defined by the first aspect of the present invention. Without limitation, this may be a bispecific immune cell engager antibody. An exemplary embodiment thereof is a bispecific T-cell engager (BiTE) antibody, optionally wherein, in addition to the region that comprises the binding molecule of the first aspect of the present invention which has binding specificity for the ECD3 region of the US28 protein, the BiTE antibody further comprises a T-cell engaging domain, such as a CD3-binding domain.

[0192] In a further embodiment, the one or more agents to be used in accordance with the eleventh aspect of the present invention may, for example, be (or include) a conjugate according to the eighth aspect of the present invention and / or the tenth aspect of the present invention, such as conjugate that is an antibody-drug conjugate ("ADC"), or a conjugate that comprises radioactive moiety, such as conjugate that is suitable for use in radioimmunotherapy ("RIT").

[0193] In another embodiment, the one or more agents to be used in accordance with the eleventh aspect of the present invention may, for example, be (or include) a cell (or population of cells, for example a homogeneous population of cells) wherein the or each cell comprises a CAR according to the fourth aspect of the present invention. Said cell or population of cells is typically isolated and / or formulated for administration to a subject. The or each cell may, for example, comprise a CAR according to the first aspect of the present invention and / or a nucleic acid encoding said CAR, optionally wherein said nucleic acid is a nucleic acid or vector as defined by the second or third aspects of the present invention, respectively. The or each cell may, for example, be a cell according to the fourth aspect of the present invention. Without limitation, said cell or cells may, for example, be selected from the group consisting of: a T cell, natural killer (NK) cell, and a macrophage. Accordingly, the cell may optionally be a CAR-T cell, a CAR-NK cell or a CAR-macrophage, and optionally, when the cell is a CAR-T cell, then for example the T-cell may be selected from the group consisting of CD8+T cells, CD4+ T cells, effector T cells, helper T cells, memory T cells, cytotoxic T lymphocytes (CTLs) , EBV-specific T cell receptor (TCR) or y6-T cell subtypes.

[0194] Optionally, in accordance with the eleventh aspect of the present invention, the subject may be administered a further substance, such as a further therapeutic, prophylactic, diagnostic, prognostic, or theragnostic substance, and optionally wherein the further substance may be administered separately, sequentially or simultaneously with the, or each of the one or more agents.

[0195] Accordingly, the eleventh aspect of the present invention also provides a method of treating a subject in need thereof, by administering to the subject a therapeutic, prophylactic, diagnostic, prognostic, or theragnostic substance, wherein the subject is also treated separately, sequentially (for example, before, or after), or simultaneously, with the, or each of the one or more agents.

[0196] In the embodiment in which the treatment is simultaneous, then the one or more agents to be used in accordance with the eleventh aspect of the present invention may be formulated and / or administered in combination with the therapeutic, prophylactic, diagnostic, prognostic, or theragnostic substance; or may be formulated separately but administered simultaneously as two separate formulations.

[0197] For example, in one embodiment, the disease or condition to be treated may be a form of cancer (such as one or more forms of cancer as disclosed above), and the additional therapeutic, prophylactic, diagnostic, prognostic, or theragnostic substance may be targeted to the cancer.

[0198] A twelfth aspect of the present invention provides an agent for use in medicine, wherein the agent is selected from the group consisting: i. a binding molecule according to the first aspect of the present invention, II. a functional fragment of said binding molecule as defined by the first aspect of the present invention, ill. an isolated binding molecule according to the seventh aspect of the present invention, iv. a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the second aspect of the present invention, v. a vector according to the third aspect of the present invention, vi. a cell according to the fourth aspect of the present invention, vii. a conjugate according to the eighth aspect of the present invention, and viii. an isolated conjugate according to the tenth aspect of the present invention.

[0199] Said agent may, for example, be an agent as define above, in respect of the eleventh aspect of the present invention.

[0200] A thirteenth aspect of the present invention provides a peptide or polypeptide comprising, consisting essentially of, or consisting of, the sequence TKKNNQCMTDYDYLEVS (SEQ ID NO: 6), or comprising the sequence of an immunogenic fragment of SEQ ID NO: 6. Said peptide or polypeptide is not the US28 protein. Preferably the only US28-derived sequence in said peptide or polypeptide is the sequence of SEQ ID NO: 6 or the sequence of the immunogenic fragment of SEQ ID NO: 6.

[0201] A fourteenth aspect of the present invention provides a peptide or polypeptide comprising, consisting essentially of, or consisting of, the sequence TKKDNQCMTDYDYLEVS (SEQ ID NO:7), or comprising the sequence of an immunogenic fragment of SEQ ID NO: 7. Said peptide or polypeptide is not the US28 protein. Preferably the only US28-derived sequence in said peptide or polypeptide is the sequence of SEQ ID NO: 7 or the sequence of the immunogenic fragment of SEQ ID NO: 7.

[0202] An immunogenic fragment of the reference sequence SEQ ID NO: 6 or 7, in accordance with the thirteenth or fourteenth aspect of the present invention, respectively, comprises less than the full sequence of the reference sequence, and preferably comprises at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 consecutive amino acids of the reference sequence.

[0203] In one embodiment, an immunogenic fragment of the peptide or polypeptide of the thirteenth and / or fourteenth aspect of the present invention, comprises, consists essentially of, or consists of, a sequence that is common to, and present within, both of SEQ ID NO: 6 and SEQ ID NO: 7.

[0204] Also provided by the thirteenth and fourteenth aspects of the present invention are peptides or polypeptides comprising, consisting, or consisting essentially of, an immunogenic fragment or variant of a reference sequence selected from TKKNNQCMTDYDYLEVS (SEQ ID NO: 6), or TKKDNQCMTDYDYLEVS (SEQ ID NO: 7), respectively, wherein the immunogenic fragment or variant comprises the sequence of the epitope within ECD3 of US28 that is bound by any of antibodies 1D3, 1C10, 1A10, 1G9 and / or 1E8, as described herein (by which is included also an scFv comprising a VH polypeptide sequence having the VH sequence of 1D3, 1C10, 1A10, 1G9 and / or 1E8, as defined by SEQ ID Nos: 12, 104, 122, 68 and 88, respectively, and a VL polypeptide sequence having the VL sequence of 1D3, 1C10, 1A10, 1G9 and / or 1E8 as defined by SEQ ID Nos: 18, 108, 126, 72 and 92, respectively). Preferably said peptides or polypeptides are bound specifically by 1D3, 1C10, 1A10, 1G9 and / or 1E8.

[0205] A fifteenth aspect of the present invention provides a combination of at least two distinct peptides and / or polypeptides, comprising a first peptide or polypeptide and a second peptide or polypeptide, wherein: the first peptide or polypeptide comprises a comprises, consists essentially of, or consists of, the sequence TKKNNQCMTDYDYLEVS (SEQ ID NO: 6), or an immunogenic fragment thereof, such as an immunogenic fragment as defined by the thirteenth aspect of the present invention, with the proviso that said immunogenic fragment comprises at least the 4N amino acid of SEQ ID NO: 6; and the second peptide or polypeptide comprises a comprises, consists essentially of, or consists of, the sequence TKKDNQCMTDYDYLEVS (SEQ ID NO: 7), or an immunogenic fragment thereof, such as an immunogenic fragment as defined by the fourteenth aspect of the present invention, with the proviso that said immunogenic fragment comprises at least the 4D amino acid of SEQ ID NO: 7.

[0206] A sixteenth aspect of the present invention provides a fusion protein comprising, consisting essentially of, or consisting of, a first amino acid sequence fused, either directly or via one or more linker amino acid sequences, to a second amino acid sequence, wherein the first amino acid sequence is the sequence of a peptide or polypeptide as defined by the thirteenth or fourteenth aspect of the present invention; and the second amino acid sequence is a fusion partner.

[0207] Optionally, the fusion partner is a carrier protein, such as a carrier protein that is selected to provide a fusion protein that is suitable for immunisation and generation of antibodies against the first amino acid sequence. For example, the carrier protein may be selected from the group consisting of keyhole limpet hemocyanin (KLH), HSA (human serum albumin), BSA (bovine serum albumin), OVA (ovalbumin), tetanus toxoid (TT), diphtheria toxoid (DT), a genetically modified cross-reacting material (CRM) of diphtheria toxin, meningococcal outer membrane protein complex (OMPC) and H. influenzae protein D (HiD).

[0208] A seventeenth aspect of the present invention provides a combination of at least two distinct fusion proteins, comprising a first fusion protein, and a second fusion protein, wherein: the first fusion protein according to the sixteenth aspect of the present invention comprises, as the first amino acid sequence of the first fusion protein, a sequence that comprises a comprises, consists essentially of, or consists of, the sequence TKKNNQCMTDYDYLEVS (SEQ ID NO: 6), or an immunogenic fragment or variant thereof, with the proviso that said immunogenic fragment or variant includes the 4N amino acid of SEQ ID NO: 6; and the second fusion protein according to the sixteenth aspect of the present invention comprises, as the first amino acid sequence of the second fusion protein, a sequence that comprises, consists essentially of, or consists of, the sequence TKKDNQCMTDYDYLEVS (SEQ ID NO: 7), or an immunogenic fragment or variant thereof, with the proviso that said immunogenic fragment or variant includes the 4D amino acid of SEQ ID NO: 7.

[0209] An eighteenth aspect of the present invention provides a conjugate, comprising a moiety conjugated to a peptide or polypeptide as defined by either or both of the thirteenth and fourteenth aspects of the present invention, or to a fusion protein as defined by the sixteenth aspect of the present invention.

[0210] The moiety of said conjugate may be conjugated directly to the peptide or polypeptide as defined by either or both of the thirteenth and fourteenth aspects of the present invention, or to the fusion protein as defined by the sixteenth aspect of the present invention. Alternatively, the moiety of said conjugate may be conjugated indirectly, such as via a linker, to the peptide or polypeptide as defined by either or both of the thirteenth and fourteenth aspects of the present invention, or to the fusion protein as defined by the sixteenth aspect of the present invention.

[0211] Optionally, the moiety may be a carrier, for example a carrier protein, such as a carrier selected from KLH (keyhole limpet hemocyanin), HSA (human serum albumin), BSA (bovine serum albumin), OVA (ovalbumin), tetanus toxoid (TT), diphtheria toxoid (DT), a genetically modified cross-reacting material (CRM) of diphtheria toxin, meningococcal outer membrane protein complex (OMPC) and H. influenzae protein D (HiD).

[0212] A nineteenth aspect of the present invention provides a combination of at least two distinct conjugates, wherein the combination comprises: a first conjugate according to the eighteenth aspect of the present invention, wherein the first conjugate comprises, consists essentially of, or consists of, a moiety conjugated to a peptide or polypeptide, wherein the peptide or polypeptide comprises, consists essentially of, or consists of, the sequence TKKNNQCMTDYDYLEVS (SEQ ID NO: 6), or an immunogenic fragment or variant thereof, with the proviso that said immunogenic fragment or variant includes the 4N amino acid of SEQ ID NO: 6; and a second conjugate according to the eighteenth aspect of the present invention, wherein the second conjugate comprises, consists essentially of, or consists of, a moiety conjugated to a peptide or polypeptide, wherein the peptide or polypeptide comprises, consists essentially of, or consists of, the sequence TKKDNQCMTDYDYLEVS (SEQ ID NO: 7), or an immunogenic fragment or variant thereof, with the proviso that said immunogenic fragment or variant includes the 4D amino acid of SEQ ID NO: 7. A twentieth aspect of the present invention provides a method of producing a conjugate according to the nineteenth aspect of the present invention, the method comprising the steps of:

[0213] (a) providing a peptide or polypeptide as defined by either or both of the thirteenth and / or fourteenth aspects of the present invention, a combination thereof as defined by the fifteenth aspect of the present invention, or a fusion protein as defined by the sixteenth aspect of the present invention; and

[0214] (b) conjugating a moiety thereto.

[0215] A twenty-first aspect of the present invention provides a method of producing a combination of at least two distinct conjugates as defined by the nineteenth aspect of the present invention.

[0216] In one embodiment, the method comprising the steps of: (a) providing or producing the first conjugate, as defined by the eighteenth aspect of the present invention, by a method according to the twentieth aspect of the present invention; (b) providing or producing the second conjugate, as defined by the eighteenth aspect of the present invention, by a method according to the twentieth aspect of the present invention (wherein the first and second conjugates are distinct); and (c) combining the first and second conjugates, thereby to form a combination according to the nineteenth aspect of the present invention.

[0217] In another embodiment, the method comprising the steps of: (a) providing a combination of at least two distinct peptides and / or polypeptides according to fifteenth aspect of the present invention, or a combination of at least two distinct fusion proteins according to the seventeenth aspect of the present invention; and (b) conjugating a moiety to the combination of at least two distinct peptides and / or polypeptides according to fifteenth aspect of the present invention, or to the combination of at least two distinct fusion proteins according to the seventeenth aspect of the present invention, thereby to form a combination according to the nineteenth aspect of the present invention.

[0218] The methods of the twentieth or twenty-first aspect of the present invention optionally comprise the step of isolating the thus-produced conjugate or combination of conjugates.

[0219] A twenty-second aspect of the present invention provides an isolated conjugate, or combination of conjugates, that is obtained, or obtainable, by the method of any of twentieth or twenty-first aspect of the present invention, optionally, wherein the isolated conjugate is further formulated for administration to a subject.

[0220] A twenty-third aspect of the present invention provides nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, wherein the nucleic acid molecule comprises, or the combination of multiple distinct nucleic acid molecules collectively comprises, one or more nucleic acid sequences that, individually or in combination, encode one or more peptides and / or polypeptides according to either or both of the thirteenth and / or fourteenth aspects of the present invention, a combination of at least two distinct peptides and / or polypeptides according to the fifteenth aspect of the present invention, a fusion protein according to the sixteenth aspect of the present invention, and / or a combination of at least two distinct fusion proteins according to the seventeenth aspect of the present invention.

[0221] The, or each, nucleic acid molecule according to the twenty-third aspect of the present invention may, for example, be each independently selected from a DNA or RNA molecule. The, or each, nucleic acid molecule according to the twenty-third aspect of the present invention may, for example, be each independently selected from a single-stranded or a double-stranded nucleic acid molecule.

[0222] A twenty-fourth aspect of the present invention provides a vector comprising a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the twenty-third aspect of the present invention. Any vector may be used, although without limitation, said vector may optionally be selected from the group consisting of a retroviral vector, a plasmid, a lentivirus vector, and an adenoviral vector.

[0223] A twenty-fifth aspect of the present invention provides a cell comprising the nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the twenty-third aspect of the present invention, or the vector according to the twenty-fourth aspect of the present invention. Optionally, the cell expresses one or more peptide or polypeptide selected from either or both of the thirteenth and / or fourteenth aspects of the present invention, a combination of at least two distinct peptides and / or polypeptides according to the fifteenth aspect of the present invention, a fusion protein according to the sixteenth aspect of the present invention, and / or a combination of at least two distinct fusion proteins according to the seventeenth aspect of the present invention.

[0224] A twenty-sixth aspect of the present invention provides a cell that is exposed to, and / or comprising, one or more peptide or polypeptide selected from either or both of the thirteenth and / or fourteenth aspects of the present invention, a combination of at least two distinct peptides and / or polypeptides according to the fifteenth aspect of the present invention, a fusion protein according to the sixteenth aspect of the present invention, and / or a combination of at least two distinct fusion proteins according to the seventeenth aspect of the present invention, a conjugate according to eighteenth aspect of the present invention, a combination of at least two distinct conjugates according the nineteenth aspect of the present invention, a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the twenty-third aspect of the present invention, and / or a vector according to the twenty-fourth aspect of the present invention.

[0225] A twenty-seventh aspect of the present invention provides a method of isolating and / or enriching cells comprising a T cell receptor (TCR) with specificity to an epitope in ECD3 of US28 (e.g. naturally occurring T cells, or recombinant cells expressing a CAR according to the first aspect of the present invention, for example CAR T-cells, CAR NK-cells and / or CAR-macrophages), wherein the method comprises the step of using one or more agents to isolate and / or enrich cells with binding specificity to one or both of the sequences of SEQ ID Nos: 6 and / or 7, wherein the one or more agents is or are selected from the group consisting of a peptide or polypeptide according to either or both of the thirteenth and / or fourteenth aspects of the present invention, a combination of at least two distinct peptides and / or polypeptides according to the fifteenth aspect of the present invention, a fusion protein according to the sixteenth aspect of the present invention, and / or a combination of at least two distinct fusion proteins according to the seventeenth aspect of the present invention, a conjugate according to eighteenth aspect of the present invention, and / or a combination of at least two distinct conjugates according the nineteenth aspect of the present invention.

[0226] In one embodiment of the method of the twenty-seventh aspect of the present invention, the sequences can be formulated as an MHC tetramer, for example a Class I MHC tetramer for antigen-specific CD8+T cells detection, a Class II MHC tetramer for antigen-specific CD4+T cells detection, or a fluorophore-labelled tetramer for flow cytometry or fluorescence microscopy. Optionally, the T-cell may be selected from the group consisting of CD8+T cells, CD4+ T cells, effector T cells, helper T cells, memory T cells, cytotoxic T lymphocytes (CTLs), EBV-specific T cell receptor (TCR) or y6-T cell subtypes.

[0227] A twenty-eighth aspect of the present invention provides an MHC tetramer comprising a peptide or polypeptide according a peptide or polypeptide according to either or both of the thirteenth and / or fourteenth aspects of the present invention, a combination of at least two distinct peptides and / or polypeptides according to the fifteenth aspect of the present invention, optionally wherein the or each peptide comprises or corresponds to SEQ ID NO:6 or SEQ ID NO:7, or an immunogenic fragment of either or both, for example wherein the MHC tetramer is a Class I MHC tetramer for antigen-specific CD8+T cells detection, a Class II MHC tetramer for antigen-specific CD4+T cells detection, a fluorophore-labelled tetramer for flow cytometry or fluorescence microscopy. The MHC tetramer may further be used in isolating and / or enriching cells comprising a T cell receptor (TCR) with specificity to an epitope in ECD3 of US28, for example, a T-cell selected from the group consisting of CD8+T cells, CD4+T cells, effector T cells, helper T cells, memory T cells, cytotoxic T lymphocytes (CTLs), EBV-specific T cell receptor (TCR) or y6-T cell subtypes and / or a recombinant cell expressing a CAR according to the first aspect of the present invention, for example a CAR T-cell, CAR NK-cell and / or CAR-macrophage.

[0228] A twenty-ninth aspect of the present invention provides a vaccine composition suitable for use in vaccinating against, reducing the risk of, preventing, or combating a disease or condition associated with human cytomegalovirus (HCMV). The vaccine may be an active or passive vaccine. An active vaccine according to the twenty-ninth aspect of the present invention may trigger an immune response directed to an epitope present within ECD3 of a US28 protein of HCMV. A passive vaccine according to the twenty-ninth aspect of the present invention may be a composition that provides an immune response directed to an epitope present within ECD3 of a US28 protein of HCMV. As discussed above in the context of other aspects of the present invention, the ECD3 of the US28 protein comprises, consists essentially of, or consist of, an amino acid sequence presented in the US28 protein encoded by a strain of HCMV at positions corresponding to positions 167 to 183 of the US28 protein encoded by the DB strain of human cytomegalovirus (HCMV) as set forth in SEQ ID NO: 5.

[0229] In some embodiments, the vaccine composition of the twenty-ninth aspect of the present invention triggers and / or provides an immune response:

[0230] (a) to one or more epitopes present entirely within extracellular domain 3 (ECD3) of the US28 protein of HCMV;

[0231] (b) to one or more linear epitopes within ECD3 of the US28 protein;

[0232] (c) to one or more epitopes within ECD3 of a US28 protein of HCMV that is an epitope, or are epitopes, present in identical form in both the 4D-variant strains and 4N-variant strains of HCMV, wherein the 4D-variant strain of HCMV encodes a US28 protein comprising an ECD3 having the sequence of TKKDNQCMTDYDYLEVS (SEQ ID NO: 7) and wherein the 4N-variant strain of HCMV encodes a US28 protein comprising an ECD3 having the sequence of TKKNNQCMTDYDYLEVS (SEQ ID NO: 6); and / or

[0233] (d) wherein the immune response that is triggered or provided by the vaccine is HCMV strain agnostic to the 4D-variant strains and 4N-variant strains of HCMV, and triggers and / or provides an immune response that is directed to one or more of the 4D-variant HCMV strains selected from Towne, VR1814, TB40 / E, Merlin, JP, Ad 169, VHL / E, BL, AF1 and DAVIS and is also directed to one or more of the 4N-variant HCMV strains selected from Toledo, TR and DB.

[0234] Said vaccine composition may be a passive vaccine, and / or optionally comprise: (a) one or more binding molecules according to the first aspect of the present invention, (b) one or more functional fragments of said one or more binding molecules as defined by the first aspect of the present invention, (c) one or more isolated binding molecules according to the seventh aspect of the present invention, (d) one or more nucleic acid molecules, or combination of multiple distinct nucleic acid molecules, according to the second aspect of the present invention, (e) one or more vectors according to the third aspect of the present invention, (f) one or more cells according to the fourth aspect of the present invention, (g) one or more conjugates according to the eighth aspect of the present invention, and / or (h) one or more isolated conjugates according to the tenth aspect of the present invention.

[0235] Alternatively, said vaccine composition may be an active vaccine, and / or optionally comprise:

[0236] (a) one or more peptides or polypeptides according to either or both of the thirteenth and / or fourteenth aspects of the present invention, a combination of at least two distinct peptides and / or polypeptides according to the fifteenth aspect of the present invention, a fusion protein according to sixteenth aspect of the present invention, a combination of at least two distinct fusion proteins according to the seventeenth aspect of the present invention, a conjugate according to the eighteenth aspect of the present invention, and / or a combination of at least two distinct conjugates according to the nineteenth aspect of the present invention;

[0237] (b) one or more nucleic acid molecules, or combination of multiple distinct nucleic acid molecules, according to the twenty-third aspect of the present invention, and / or the vector according to the twenty-fourth aspect of the present invention; and / or

[0238] (c) a cell, such as an antigen-presenting cell (e.g. a dendritic cell), or a homogeneous or heterogeneous population of said cells, wherein the or each of said cells is loaded with one or more of the following : a peptide or polypeptide according to either or both of the thirteenth and / or fourteenth aspects of the present invention, a combination of at least two distinct peptides and / or polypeptides according to the fifteenth aspect of the present invention, a fusion protein according to sixteenth aspect of the present invention, a combination of at least two distinct fusion proteins according to the seventeenth aspect of the present invention, a conjugate according to the eighteenth aspect of the present invention, a combination of at least two distinct conjugates according to the nineteenth aspect of the present invention, one or more nucleic acid molecules, or combination of multiple distinct nucleic acid molecules, according to the twenty-third aspect of the present invention, and / or the vector according to the twenty-fourth aspect of the present invention.

[0239] A thirtieth aspect of the present invention provides a method of vaccinating against, reducing the risk of, preventing, and / or combating a disease or condition associated with HCMV, the method comprising administering to a subject a vaccine according to the twenty-ninth aspect of the present invention. The thirtieth aspect of the present invention provides a vaccine according to the twenty-ninth aspect of the present invention for use in vaccinating against, reducing the risk of, preventing, and / or combating a disease or condition associated with HCMV in a subject.

[0240] The thirtieth aspect of the present invention provides for the use of a vaccine according to the twenty-ninth aspect of the present invention in the manufacture of a medicament for vaccinating against, reducing the risk of, preventing, and / or combating a disease or condition associated with HCMV.

[0241] Said disease or condition associated with HCMV may, for example, be a disease or condition associated with HCMV as disclosed above in the context of the eleventh aspect of the present invention. Optionally, the disease or condition is a latent HCMV infection, or is a disease or condition associated with a latent HCMV infection.

[0242] In some embodiments, the method of vaccinating against, reducing the risk of, preventing, and / or combating a disease or condition associated with HCMV may comprise administering the vaccine to the subject only once.

[0243] In other embodiments, the method of vaccinating against, reducing the risk of, preventing, and / or combating a disease or condition associated with HCMV may comprise administering the vaccine to the subject vaccine twice or multiple times. For example, in the embodiment in which the vaccine is an active vaccine, it may be appropriate to separately administer a primary dose, and a subsequent booster dose, to the subject.

[0244] A thirty-first aspect of the present invention provides a method of assessing one or more biological conditions and / or biological characteristics of a subject and / or of ex vivo biological material, wherein the method comprises: (a) contacting the subject and / or the ex vivo biological material with a binding molecule, for example as defined by the first aspect of the present invention, or a conjugate, for example as defined by the eighth or tenth aspect of the present invention; and (b) making an assessment of the subject and / or the ex vivo biological material based on a direct and / or indirect measurement of the binding of the binding molecule or conjugate to the subject and / or the ex vivo biological material.

[0245] In some embodiments, the method of assessing (which can include diagnosing) one or more biological conditions and / or biological characteristics of a subject and / or of ex vivo biological material is by In Situ Hybridisation (ISH) for the specific detection of one or more nucleic acid sequences encoded by HCMV, most preferably wherein said ISH does not detect nucleic acid sequences encoded by a healthy (i.e. not infected by HCMV) subject and / or a healthy ex vivo biological material. By way of non-limiting example, the ISH may use nucleic acid sequences as discussed in the Examples of the present application. In some embodiments, the biological condition is cancer and / or the biological characteristics are related to cancer. In some embodiments, the cancer is selected from one or more of the cancers specified herein, optionally wherein the cancer is not glioblastoma. In some embodiments, the cancer is selected from the group consisting of: breast cancer (for example HER2+ breast cancer, or triple negative breast cancer), astrocytoma, glioblastoma, adrenal cortical cancer, kidney cancer, cardiac sarcoma, liver cancer, and vascular smooth muscle cancers; optionally wherein the cancer is not glioblastoma.

[0246] In some embodiments, the method of assessing (which can include diagnosing) one or more biological conditions and / or biological characteristics of a subject and / or of ex vivo biological material is by immunohistochemistry (IHC) for the specific detection of one or more protein sequences encoded by HCMV, most preferably wherein said IHC does not detect protein sequences encoded by a healthy (i.e. not infected by HCMV) subject and / or a healthy ex vivo biological material. In some embodiments, the IHC uses a binding molecule that allows for the specific detection of one or more protein sequences only expressed, or only surface expressed, by a latent HCMV infection. In some embodiments, the IHC uses a binding molecule that allows for the specific detection of one or more protein sequences only expressed, or only surface expressed, by a lytic HCMV infection. In some embodiments, the IHC uses a binding molecule that allows for the specific detection of one or more protein sequences expressed, for example surface expressed, by a both lytic and latent HCMV infections. In some embodiments, the IHC may be performed (e.g. using biological tissue in which the cells have not been permeabilised) to allow only for the detection of cell surface- expressed proteins. In other embodiments, the IHC may be performed (e.g. using biological tissue in which the cells have been permeabilised) to allow for the detection of intracellular proteins. By way of non-limiting examples, the IHC may use an antibody-based molecule for the specific detection of one or more protein sequences encoded by HCMV, as discussed in the Examples of the present application and / or using one or more of the binding molecules of the present invention. In some embodiments, the biological condition is cancer and / or the biological characteristics are related to cancer. In some embodiments, the cancer is selected from one or more of the cancers specified herein, optionally wherein the cancer is not glioblastoma. In some embodiments, the cancer is selected from the group consisting of: breast cancer (for example HER2+ breast cancer, or triple negative breast cancer), astrocytoma, glioblastoma, adrenal cortical cancer, kidney cancer, cardiac sarcoma, liver cancer, and vascular smooth muscle cancers; optionally wherein the cancer is not glioblastoma.

[0247] A subject and / or living ex vivo biological material in which HCMV infection has been positively identified by the thirty-second aspect of the present invention can be an exemplary subject and / or material that can be treated in accordance with the other aspects of the present invention as described herein.

[0248] A thirty-second aspect of the present invention provides a method of combating a HCMV infection (such as a latent HCMV infection and / or a lytic HCMV infection and / or a multi-strain HCMV infection) in living ex vivo biological material, the method comprising contacting the living ex vivo biological material with any one or more agents selected from the group consisting of: i. a binding molecule according to the first aspect of the present invention,

[0249] II. a functional fragment of said binding molecule as defined by the first aspect of the present invention, ill. an isolated binding molecule according to the seventh aspect of the present invention, iv. a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to the second aspect of the present invention, v. a vector according to the third aspect of the present invention, vi. a cell according to the fifth aspect of the present invention, vii. a conjugate according to the eighth aspect of the present invention, and viii. an isolated conjugate according to the tenth aspect of the present invention.

[0250] The thirty-second aspect of the present invention also provides living ex vivo biological material that is obtained, or obtainable, by the method of this aspect.

[0251] In one embodiment of the method of the thirty-second aspect of the present invention, and / or the living ex vivo biological material obtained, or obtainable, thereby, the ex vivo living biological material comprises, consists essentially of, or consists of, living ex vivo biological material selected from the group that includes: one or more types of ex vivo cells; one or more types of ex vivo cell cultures; one or more types of ex vivo tissues; one or more types of ex vivo tissue cultures; one or more types of ex vivo organoids; one or more types of ex vivo organoid cultures; one or more types of ex vivo organs; and / or one or more types of ex vivo organ cultures.

[0252] A thirty-third aspect of the present invention provides a method of treating a subject in need thereof, comprising administering ex vivo living biological material as defined by the thirty-second aspect of the present invention, to the subject.

[0253] For example, the method may be a method of transplantation of the ex vivo living biological material, such as an organ or tissue transplant. Said method can be used to prevent, or reduce the risk of, the transmission of and HCMV infection, or a disease or condition associated with an HCMV infection in the recipient of the transplant. A thirty-fourth aspect of the present invention provides a method of screening for a binding molecule having binding specificity and / or binding affinity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of human cytomegalovirus (HCMV), the method comprising:

[0254] (a) providing one or more peptides corresponding an amino acid sequence present in ECD3 of the US28 protein,

[0255] (b) providing one or more candidate binding molecules;

[0256] (c) determining the binding specificity and / or binding affinity of one or more candidate binding molecules to the one or more peptides.

[0257] A thirty-fifth aspect of the present invention provides a method of producing a composition that comprises multiple copies of a binding molecule, said method comprising causing the reproduction of a selected candidate binding molecule that has been selected in accordance with the method of the thirty-fourth aspect of the present invention.

[0258] A thirty-sixth aspect of the present invention provides a method of assessing a selected candidate binding molecule that has been selected in accordance with the method of the thirty-fourth aspect of the present invention and / or produced in accordance with the method of the thirty-fifth aspect of the present invention, said method comprising and identifying the structure(s) within the selected candidate binding molecule that provides its binding characteristics (in particular, the binding specificity and / or binding affinity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of human cytomegalovirus (HCMV)), for example, by identifying the, or each, CDR sequence in a selected candidate binding molecule that is an antibody or CAR.

[0259] A thirty-seventh aspect of the present invention provides a method of producing a composition that comprises multiple copies of a binding molecule having binding specificity and / or binding affinity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of human cytomegalovirus (HCMV), wherein said binding molecule comprises the, or each, of the structure(s) (e.g. CDR sequences) that have been identified within a selected candidate binding molecule as providing its binding characteristics, in accordance with the method of the thirty-sixth aspect of the present invention, said method comprising causing the reproduction of the binding molecule.

[0260] The thirty-seventh aspect of the present invention further provides a composition of binding molecule obtained by the same aspect.

[0261] DESCRIPTION OF THE FIGURES

[0262] Figure 1. Western blot analysis was used to verify the US28 protein expression in the transformed CHO-US28-A1 cells by using an anti-HIS Ab. From the left, on the first line: the ladder, on the second line: protein extract from the CHO control cells, on the third line: protein extract from the CHO-US28-A1 cells and on the fourth line: the 6HIS positive control. The C-terminal part of the US28-A1 construct, which was transfected into the CHO-US28-A1 cells, contains a 6HIS tag. Staining with the anti- HIS antibody, that binds on the 6HIS tag marked out a specific band at -41 kDa when compared with the ladder, which is equivalent with the earlier reported size of the HCMV US28 protein. This band, which is circled in the picture, was not present in the control CHO-cells, therefore verifying the expression of US28 in the transfected CHO- US28-A1 cells but not in the control CHO cells. The 6HIS positive control is positive indicating that the 6HIS Ab binds to its target as expected.

[0263] Figure 2. Flowcytometry analysis showing surface binding of the US28-13-5G6- 1D3 antibody clone on CHO-US28-A1 cells. From the left, blank control, 2nd antibody control and clone US28-13-5G6-1D3 Ab binding on the surface of the CHO-US28-A1 cells. The US28-13-5G6-1D3 Ab bound to 53,4% of the CHO-US28-A1 cells, whereas the blank and secondary antibody controls showed low binding: 0.11% and 1.63% respectively.

[0264] Figure 3. The positive surface binding of the clone US28-13-5G6-1D3 on CHO- US28-A1 cells was further validated by gradient dilution in FACS analysis. From the left, the clone US28-13-5G6-1D3 Ab surface binding to the CHO-US28-A1 cells in dilutions 1 :20, 1 : 50, 1:200 (w / v).

[0265] Figure 4. The positive surface binding of the clone US28-13-5G6-1D3 on CHO- US28-A1 cells was compared with the US28 negative CHO control cells. The clone US28-13-5G6-1D3 Ab bound 15-fold more to the surface of the US28 positive CHO- US28-A1 cells than to the US28 negative CHO cells in 1 : 50 (w / v).

[0266] Figure 5. The original ELISA analysis showed equal qualitative binding of the US28-13-5G6-1D3 antibody clones on both Bio-Peptide-1 (US28 ECD3 genotype 4N) and Bio-Peptide-2 (US28 ECD3 genotype 4D). Since no other mutations in the ECD3 has been observed, these results indicate that the binding of the US28-13-5G6-1D3 Ab to its target is HCMV strain agnostic.

[0267] Figure 6. Qualitative ELISA analysis showing binding of the recombinant antibody product US28-13-5G6-1D3 rAb to Bio-Peptides 1 and 2. These results show strong and equal binding of the generated antibody to both genetic variants of US28 ECD3 confirming that the recombinant antibody binds well to its target and maintains the HCMV strain agnostic binding properties.

[0268] Figure 7. Surface binding of 13-5G6-1D3 rAb on CHO-US28-A1 cells, and on respective control CHO cells were validated by using FACS analysis. The bars showing results for CHO cells are colored with dark grey whereas the bars showing results for CHO-US28-A1 cells are colored with pale grey color. From the left, the first bar shows measurement of the blank control with no antibody for the CHO-US28-A1 and CHO cells, respectively; the second bar from the left shows surface binding of the antimouse IgG-Alexa 488 secondary Ab alone on both cell types; the third bar from the left shows surface binding of 13-5G6-1D3 rAb in 1 : 10 dilution (w / v) on both cell types; the fourth bar from the left shows 13-5G6-1D3 rAb surface binding in 1 : 50 dilution (w / v) on CHO-US28-A1 cells only; and the fifth bar from the left shows the surface binding of 13-5G6-1D3 rAb in 1 : 100 dilution (w / v) on CHO-US28-A1 cells only. The y- axis shows the binding to the percent of cells. Based on these results, the optimal staining dilution for 13-5G6-1D3 rAb is 1 :50 w / v (the third bar from the left). The surface binding of the 13-5G6-1D3 rAb to CHO cells was only measured in 1: 10 dilution (w / v) and was 0.77% after removing the binding to blank and anti-mouse IgG-Alexa 488 secondary Ab controls. These results are consistent with the previous results obtained by using the antibody from 13-5G6-1D3 clone and shows highly specific surface binding of the 13-5G6-1D3 rAb on CHO-US28-A1 cells, which is >21-fold more than to the normal CHO cells.

[0269] Figure 8. Binding of the US28-13-5G6-1D3 rAb on the surface of the HCMV Adl69 infected MRC-5 cell population but not on the Mock cells were shown by FACS analysis. A. The upper row shows the surface binding of the US28-13-5G6-1D3 rAb on Mock versus HCMV Adl69 infected MRC-5 cells showing highly specific binding on the HCMV infected cells. B. To verify that the 13-5G6-1D3 rAb was binding to the surface of the HCMV infected cell population, another set of HCMV Ad 169 infected and Mock cells were permeabilized and stained with the commercial antibody MAB810X against the HCMV Major Immediate Early (IE) antigen, which is an intracellular protein known to be expressed early during the lytic HCMV infection. The MAB810X antibody stained the same HCMV infected MRC-5 cell population as the US28-13-5G6-1D3 rAb, which was not observed for non-infected cells for either antibodies, confirming that the surface binding demonstrated for the US28-13-5G6-1D3 rAb was specific for the HCMV Ad 169 infected cells.

[0270] Figure 9. Non-specific binding of the US28-13-5G6-1D3 rAb was tested by using mouse IgG isotype control instead of the primary antibody prior to incubation with Goat anti-mouse IgG-Alexa 488. The results showed a shift in staining of the HCMV infected MRC-5 cells by the US28-13-5G6-1D3 rAb compared with the isotype control, while staining of uninfected cells (Mock) were similar to the IgG isotype control. Taken together, these results show that the US28-13-5G6-1D3 rAb binds specifically (~16- fold greater binding) to the surface of the lytically HCMV infected cells as compared with the uninfected cells.

[0271] Figure 10. Binding of US28-13-5G6-1D3 rAb to primary PBMCs from three HCMV seropositive individuals was investigated by using the same flowcytometry protocol as in earlier studies. The bars in the figure show the relative surface binding of the US28-13-5G6-1D3 rAb on PBMCs from the respective donors. The results showed surface binding of the US28-13-5G6-1D3 rAb on 18.37%, 3.57% and 5.25% of the total PBMCs from the three individuals, respectively. The relative proportion of PBMCs expressing certain different cell surface markers from the same individuals are listed on under the bars; totals exceed 100% because some of the same cells express more than one of these cell surface markers (however, they are not measured during the same experiment than the US28-13-5G6-1D3 rAb binding). Of the studied PBMCs, only the CD11 + , CD14+ and CD16+ positive cells can be carriers of the HCMV, and these surface markers can partly overlap in different mononuclear cells. The population of mononuclear cells latently infected with HCMV often adds up to about 15% of total PBMCs (although there can be some considerable variation between individuals). Consequently, the observed surface binding of the US28-13-5G6-1D3 rAb on 18.37%, 3.57% and 5.25% of the total PBMCs in donors 1, 2 and 3, respectively, demonstrates binding to a substantial proportion of the PBMCs that can be HCMV carriers. These results are not related to the subtypes of the PBMCs, but indicate that the US28-13- 5G6-1D3 rAb can bind on the surface of latently HCMV infected cells.

[0272] Figure 11. Binding of the commercial US28 polyclonal Ab to CHO-US28-A1 cells was tested and compared with the binding of US28-13-5G6-1D3 rAb to the same cells. A. The US28-13-5G6-1D3 antibody bound to the surface of more than 50% of the CHO-US28-A1 cells (first bar from the left), while the commercial US28 Ab only bound on 10% of the cells (second bar from the left). B. We then tested the commercial US28 Ab on HCMV Ad 169 infected MRC-5 cells, where it showed specific binding to the HCMV infected population. However, the commercial US28 polyclonal Ab also stained the noninfected Mock cells about two-fold compared to the IgG isotype control indicating nonspecific binding on the surface of non-infected MRC-5 cells. The binding specificity of the commercial US28 Ab to HCMV Adl69 positive MRC-5 cells was only 1.7 when compared with the US28 negative Mock MRC-5 cells when the signal from IgG isotype was subtracted.

[0273] Figure 12. US28-13-5G5-1D3 binding in 1 :400 dilution (w / v) to HCMV-infected human tissues were studied by using immunohistochemistry analysis (IHC). A. Staining of the HCMV infected lung tissue with the US28-13-5G6-1D3 rAb demonstrated specificity for the HCMV infected alveolar and endothelial cells. The arrows (in the non-colour version of this figure) mark the US28-13-5G6-1D3 rAb cytoplasmic staining on typical HCMV infected alveolar cells with the characteristic cytomegalo effect in the HCMV positive control slide. B. 13-5G6-1D3 shows positive US28 staining for macrophages, but negative staining for the alveolar cells in normal lung tissue. C. The same macrophages show positive HCMV DNA in the same lung biopsy, whereas there are no HCMV DNA signals in normal alveolar cells, which is coherent with the US28 staining in the same sample. D. Staining of the HCMV infected lung tissue with the MAB810R antibody demonstrated specificity for the HCMV infected alveolar and endothelial cells. The arrows (in the non-colour version of this figure) mark the MAB810R antibody nuclear staining on typical HCMV infected cells with the characteristic cytomegalo effect in the HCMV positive control slide. E. The IE staining is negative for the alveolar HCMV positive macrophages in normal lung tissue. The normal alveolar cells show negative staining as well. F. IgG was used as negative control antibody and showed negative staining for the same lung tissues. Colour versions of Figure 12 are also provided alongside the non-colour versions.

[0274] Figure 13. The HCMV US28 expression was studied by immunohistochemistry (IHC) and in situ hybridisation (ISH) in a breast cancer cohort. A. The US28-13-5G6- 1D3 rAb showed strong cytoplasmic staining (3+) for the cancer cells (marked with arrows in the non-colour version of this figure) in a triple negative breast cancer (TNBC) sample. Most tumor cells in this sample stained positively for the US28-13-5G6-1D3 rAb (brown colour). B. The same sample showed some dot-like positive cytoplasmic staining in a few cells within the whole sample for the anti-IE MAB810R antibody. C. The ISH analysis shows multiple HCMV DNA signals in many nuclei of the same tumor cells, which is typical for many triple negative and HER2 positive tumors in our material. The arrows point out some positive HCMV DNA signals in the figure. No HCMV DNA was seen in the cytoplasm of the cells. D. The negative control IgG Ab shows negative staining. A colour version of Figure 13 is provided on the page after the non-colour version.

[0275] Figure 14. A. Glandular metastasis from HER2 positive breast cancer shows positive intermediate cytoplasmic staining (2+) for the US28-13-5G6-1D3 rAb (brown colour) in all tumor cells. B. Most cancer cells in the same sample were negative for the MAB810R staining. Only one or two cells in the whole sample had dot-like positive, cytoplasmic staining for the MAB810R Ab (not shown in the figure). C. The ISH analysis shows many positive nuclear HCMV DNA signals in the same tumor cells. The arrows point out such positive HCMV DNA signals. The HCMV DNA was not seen in the cytoplasm of the tumor cells. D. The negative control anti-IgG Ab staining for the same sample was negative. A colour version of Figure 14 is also provided.

[0276] Figure 15. The US28 staining and HCMV ISH were negative in many normal adjacent tumor (NAT) breast tissues as exemplified in the figure with the sample BR1008b_J6. A. The US28-13-5G6-1D3 rAb showed negative IHC staining for the normal breast tissue. B. The MAB810R antibody showed also negative staining for the same sample. C. No nuclear or cytoplasmic HCMV DNA signals by ISH were seen for the same sample. D. The IgG staining was also negative. These results indicate that the antibodies showed consistently negative staining for the normal glandular breast tissue where there were no signs of HCMV infection. Of note, the HCMV negative sample BR1008b_J6 was placed on the same TMA slide as the samples BR1008b_D5 and BR1008b_H6, which showed strong positive staining for the US28-13-5G6-1D3 antibody and positive nuclear signals for the HCMV DNA ISH. Thus, the samples are exposed to exactly the same staining conditions and antibody / DNA probe concentrations. The positive staining seen with the US28-13-5G6-1D3 rAb was evaluated to be specific for the HCMV infected cells in both the breast cancer and normal tissues. A colour version of Figure 15 is also provided.

[0277] Figure 16. A. The IHC staining of the primary breast cancer samples with US28-13-5G6-1D3 rAb were specific and positive in ~73% of the 41 studied primary breast tumor samples. 100% of the tumors that were positive for the US28 staining were also positive for the nuclear HCMV DNA signals, and there were multiple signals in the nuclei of 12 / 41 tumors. All 10 normal adjacent tissue controls (n = 10) were negative for the cytoplasmic US28 staining in breast epithelial cells (0) and 7 / 10 were also negative for the ISH signals. B. The positive staining with the US28-13-5G6-1D3 rAb was seen in ~94% of the studied 30 breast cancer metastases and was specific for the metastatic cells. All TNBC (n = 7) and HER2 positive (n = 11) metastases had positive US28-13-5G6-1D3 staining, and the only negative samples (n = 2) were hormone receptor positive cancers. The ISH analysis showed positive nuclear HCMV DNA in 100% of the metastases. 43% of the metastatic tissues showed multiple nuclear DNA signals in the tumor cells. These samples were exclusively of TNBC or HER2+ subtype and the two exceptions had low HR+ status. The ISH results are not shown in the figure.

[0278] Figure 17. Human glioblastoma tissues were studied with the US28-13-5G6- 1D3 rAb and MAB810R antibodies by using IHC and with ISH analysis for HCMV DNA. A. The Glioblastoma grade IV brain tumor shows moderate staining for US28 in tumor cells (brown colour). B. The MAB810R antibody staining shows no IE expression in the same cancer cells. However, the positive cytoplasmic staining for MAB810R was seen in ~90% of the glioblastoma samples. The staining was present in several cells in each positive sample which was different from the breast cancer samples. However, the staining of the glia cells was weaker than for the US28-13-5G6-1D3 rAb. C. The brain tissue NAT is negative for 13-5G6-1D3 staining. D. The ISH analysis shows positive nuclear HCMV DNA signals in the same tumor cells. The arrows indicate these signals. E. The staining of the same Glioblastoma grade IV sample is negative for the IgG antibody. F. The ISH analysis is negative for the same brain tissue NAT sample. The positive glia cells are indicated with arrows in the non-colour version of these figures. A colour version of Figure 17 is provided on the page after the non-colour version. Figure 18. US28-13-5G6-1D3 staining of the human brain cancer cohort containing human astrocytoma grade 1-3 and glioblastoma grade 4 and NAT tissues. All the astrocytoma grade 1 (n = 4) stained negative (0). Of the grade 2-3 astrocytomas, 6 stained negative (0), 37 had positive cytoplasmic staining (1 + ), and 2 had moderately positive cytoplasmic staining (2+). All of the glioblastoma grade 4 (n = 19) tumors showed positive cytoplasmic staining (1 + ) for the US28-13-5G6-1D3 rAb, of which 3 were moderately positive (2+). The ISH analysis confirmed presence of nuclear HCMV DNA in tumor cells of all tumor samples. Only 3 of 10 NAT samples were HCMV DNA positive.

[0279] Figure 19. A. Flowcytometry analysis with the 28-13-5G6-1D3-PE antibody were conducted on various human primary cells to exclude general off-target binding to the surface of these cell types. The following cell lines were studied: human primary adrenal cortical cells, human primary kidney epithelial cells, human primary cardiac microvascular endothelial cells, human primary liver epithelial cells and human primary vein smooth muscle cells. US28 targeting antibody 13-5G6-1D3 showed very low surface binding, far below < 1% to all these cell types indicating that there is no off- target surface binding of the 13-5G6-1D3 against these cell types. These results support our observations on laboratory cells showing low binding to the surface of the normal, US28 negative cells. B. Original human primary cardiac cells were not available, and human cardiac tissue samples were therefore studied for staining of 13- 5G6-1D3 by immunohistochemistry. The example in the figure shows negative 13- 5G6-1D3 staining of the heart muscle tissue similar to the staining with the negative IgG antibody control. The ISH analysis did not show any indications for presence of latent HCMV infection in the human heart muscle. If there were HCMV infections present in the human heart, they were always of productive infection type. The positive control was located on the same TMA plate with the heart samples and was positive: 13-5G6-1D3 showed positive staining for the malignant pheochromocytoma. The ISH control showed also typical HCMV DNA staining in the nucleus of the tumor cells in the same biopsy indicating presence of latent HCMV infection in this tumor type. The positive 13-5G6-1D3 and HCMV ISH results are marked with arrows in the figure. A colour version of Figure 19 is also provided.

[0280] Figure 20. Some pancreatic tissues were studied to exclude general off-target binding of the 28-13-5G6-1D3 to this tissue type. The upper row shows normal pancreatic tissue with negative staining with the 13-5G6-1D3, MAB810R and IgG control antibodies. The HCMV ISH is negative indicating absence of viral HCMV DNA in the normal pancreatic tissue sample. The lower row shows pancreatic tissue sample from another patient with positive 13-5G6-1D3 and MAB810R staining and negative IgG control antibody staining (negative control). The HCMV ISH analysis shows also positive results indicating presence of cytoplasmic HCMV DNA aggregates in the same sample. Thus, the same sample is positive for both HCMV proteins US28 and IE and HCMV DNA but negative for the negative control IgG indicating specific antibody binding. The US28 protein staining is cytoplasmic as expected, the IE protein is mostly located in the cell nucleus as expected since it is a nuclear protein. The HCMV ISH shows large cytoplasmic viral DNA aggregates since the virus is packed in the cytoplasm as indicative for productive HCMV infection. These results indicate that the antibody US28-13-5G6-1D3 does not bind off-target to pancreatic tissue and that the HCMV infection present in the morphologically normal pancreatic tissue is of productive nature in contrast to tumors where it seems to be of latent character. A colour version of Figure 20 is also provided.

[0281] Figure 21. A. Absolute binding levels of a panel of antibodies, comparing the binding properties of prior-art disclosed VUN100 (monovalent, or bivalent) to US28- 13-5G6-1D3, with respect to control CHO cells and CHO cells expressing US28 encoded by different HCMV strains (DB and TB40 / E), as well to CHO cells expressing a modified US28 containing all known US28 mutations ("mutated strain"). B. Binding specificity of the same panel of antibodies to CHO cells expressing different forms of US28 compared to control CHO cells, with results expressed as a fold-increase in binding to the US28-expressing cells compared to control CHO cells. C. Impact of strain differences on the extent of absolute binding levels of the same panel of antibodies, shown as a percentage change in absolute binding levels between the binding to CHO cells expressing US28 encoded by HCMV strains DB and TB40 / E, wherein a smaller percentage change is indicative of a greater degree of strain agnostic binding. D. Percentage retention of binding specificity of the same panel of antibodies, between CHO cells expressing the forms of US28 encoded by HCMV strains DB and TB40 / E, wherein a retention of binding specificity close to 100% is indicative of strain agnostic binding specificity, whereas a more substantial difference (as demonstrated by the two VUN100 antibody samples) indicates a change in binding specificity dependent on the strain that encodes US28 (i.e. a lack of strain agnostic binding specificity). E. Percentage binding of antibodies to CHO cells expressing the different forms of US28 (as encoded by DB, TB40 / E and the mutated form), when normalised to the level of binding observed for 1D3 for each US28 form, wherein changes in the % binding for a given antibody, across the different CHO forms, is indicative of a reduction in strain agnostic binding activity compared to the lD3-mFc antibody, and shows that binding more dependent on the strain that encodes US28 (i.e. a lack of strain agnostic binding compared to the lD3-mFc antibody). Figure 22. Percentage off-target binding of various ECD3-binding molecules of the invention, in comparison with VUN100, as assessed by binding to control CHO cells.

[0282] Figure 23. Fold change in binding specificity for exemplary ECD3-binding molecules binding to US28-expressing CHO versus control CHO cells, relative to the level of specificity of VUN 100 in the same assay. A. Improved binding specificity for 1D3 compared with VUN100 (n=3). B. Improved binding specificity for 1C10 compared with VUN100 (n = 3). C. Improved binding specificity for 1A10 compared with VUN 100 (n = 3). D. Improved binding specificity for 1G4 compared with VUN100 (n = 3). E. Improved binding specificity for 1E8 compared with VUN 100 (n= l).

[0283] DETAILED DESCRIPTION OF THE INVENTION

[0284] The present invention relates to agents targeting a specific region of the US28 protein, as encoded by human cytomegalovirus (HCMV), and therapeutic, prophylactic and diagnostic approaches related thereto including but not limited to HCMV-infected cancers and other conditions associated with latent or lytic HCMV infections.

[0285] HCMV US28 protein represents excellent potential for targeting HCMV infections, including the latent reservoir, because:

[0286] (1) Certain parts of the US28 protein are expressed on the cell surface during both lytic and latent HCMV infections (Elder et al., / 'Science, 2019, 12: 13-26);

[0287] (2) US28 is a GCPR, of which trafficking to the plasma membrane allows both its direct targeting with binding molecules and its use as a transporter of payload due to its endocytosis, which is either constitutive or occurs as a result of ligand binding. GCPRs in general constitute the largest family of proteins targeted by approved drugs (Sriram & Insel, Mol Pharmacol, 2018, 93(4) : 251-258); and

[0288] (3) In contrast to many approved drugs, which target the GCPRs encoded by human DNA, the HCMV US28 is entirely encoded by viral DNA, employing therefore a highly specific drug target exclusively located in the HCMV infected but not in healthy human cells.

[0289] A. Binding Molecules to US28

[0290] When considering the design of binding molecules against US28, the inventor noted that the N-terminal domain is the ligand binding part of the US28 protein, physically extending from the plasma membrane and therefore, most likely exposed to host antibody production, immune response, and genetic selection pressure (Mozzi et al, 2020, supra). Consistently, it is also an area known to contain high inter-strain variability and mutations (Arav-Boger et al., 2002, supra). The less conserved sites are also more susceptible to develop new mutations, which may change the response to treatment, that targets these areas over time (Komatsu et al., Antiviral Res, 2014, 101 : 12-25). Thus, new mutations in HCMV genes are likely to arise in less conserved sites, which are pinpointed by the variations between different viral strains. Most structural antibodies arise most likely against the N-terminal part of the US28 protein (De Groof et al., Mol Pharm, 2019, 16(7): 3145-3156), which may therefore also be a case for the natural antibodies in human body (Elder et al, 2019, supra).

[0291] According to our knowledge, three types of HCMV high-risk oncogenic strains have so far been identified. The DB (KT95923) and BL (MW980585) clinical HCMV isolates have been recently identified to promote oncogenic molecular pathways, establish anchorage-independent growth in vitro and produce tumorigenicity in mice models, and are therefore named as high-risk oncogenic strains (Kumar et al., 2018 and Ahmad et al., 2021). In addition, Soroceanu et al. sequenced the C-terminal part of the US28 gene in 10 HCMV positive glioblastoma tumors. Alignment of the results (presented in NIHMS323374-supplement-l.pdf for the original publication) showed that all of these strains would be similar to the HCMV VHL / E clinical isolate (L20501.1). The high-risk oncogenic strains DB, BL and VHL / E all show different mutations in the US28 gene, especially in the N-terminal part (Table 3). The N-terminal part differ from DB strain in positions E18D; A19E; F25L (VHL / E) and A19D, T21A, F25L (BL). Thus, the amino acid positions 18, 19, 21 and 25 are all mutated in these high-risk oncogenic strains; and other strains are also known to have mutations at positions 8 and / or 15 (Table 3).

[0292] Due to its highly variable sequences, the inventor considered regions of the N- terminal part of the US28 protein (ECD1) as an inappropriate target for a highly specific, and strain agnostic, antibody against US28.

[0293] In contrast, the inventor determined that ECD2 and ECD3 of US28 are highly conserved between the different HCMV strains; ECD2 does not have any known mutations, whereas the inventor's analysis of the known sequences of the ECD3 from many different HCMV strains revealed the existence of only one known mutation. Of the known high-risk oncogenic strains, the DB strain is ECD3 N170N (also referred to herein as the "4N" variant form, as position 170 of the US28 protein corresponds to position 4 of ECD3), whereas the BL and VHL / E strains represent the N 170D (also referred to herein as the "4D") variants (Table 3). The inventor identified that DNA sequence encoding US28 ECD4 DNA contains multiple polymorphic sites, of which only one leads to a more common amino acid change (R267K). Of note, the high-risk oncogenic BL strain contains both ECD4 variants V250L and R267K (Table 3).

[0294] Surprisingly, the present studies demonstrated that ECD2 and the conserved parts of ECD4 protein were not appropriate immunogens in mice, whereas the applicant was able to develop a new approach to the identification of highly specific, strain agnostic, binding molecules against the ECD3 region of HCMV US28. As reported in the examples of the present application, monoclonal antibodies were shown to bind well and specifically on both genetic variants of the US28 ECD3 peptides, on US28 overexpressing US28-CHO-A1 cells, HCMV Adl69 infected MRC-5 cells, primary PBMCs from HCMV seropositive individuals, HCMV infected human lung tissue and several types of aggressive human tumors, such as oesophagus, gastric, rectum, liver, lung, pancreas, cervical cancers, malignant pheochromocytoma and locally advanced colon cancer, breast cancer and its metastasis and glioblastoma grade 4.

[0295] Accordingly, the applicant has provided highly specific, and HCMV strain agnostic, binding molecules against the US28 protein encoded by HCMV, including, but not limited to, in particular, antibodies and chimeric antigen receptors ('CARs'), and uses thereof, for example in diagnostic, prophylactic and therapeutic uses and methods related to HCMV. Also provided are HCMV vaccines and other agents suitable for use in generating said binding molecules.

[0296] A first aspect of the present invention provides a binding molecule, comprising one or more polypeptide chains, said binding molecule having binding specificity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of human cytomegalovirus (HCMV), wherein ECD3 of the US28 protein comprises an amino acid sequence presented in the US28 protein at positions corresponding to positions 167 to 183 of the US28 protein encoded by human cytomegalovirus (HCMV) as set forth in SEQ ID NO: 5. Non-limiting, but particularly preferred, examples of said binding molecule according to the first aspect of the present invention includes antibodies and chimeric antigen receptors (CARs), as discussed further below.

[0297] Epitopes:

[0298] As noted above, binding molecules according to the first aspect of the present invention have binding specificity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of human cytomegalovirus (HCMV), wherein ECD3 of the US28 protein comprises an amino acid sequence presented in the US28 protein at positions corresponding to positions 167 to 183 of the US28 protein encoded by human cytomegalovirus (HCMV) as set forth in SEQ ID NO: 5.

[0299] The epitope to which the binding molecule of the first aspect of the present invention has binding specificity may, for example, preferably be present entirely within extracellular domain 3 (ECD3) of the US28 protein of HCMV.

[0300] The epitope may, for example, be a linear epitope within (preferably entirely within) ECD3 of the US28 protein. Alternatively, the epitope may be a discontinuous and / or conformational epitope within (preferably entirely within) ECD3 of the US28 protein. The epitope may, for example, comprise or consist of 17 or fewer amino acids of the ECD3 of the US28 protein of HCMV, for example it may comprise or consist of 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3 or fewer amino acids of the ECD3 of the US28 protein of HCMV, which may optionally be consecutive amino acids in ECD3 of US28.

[0301] The amino acid sequence presented in the US28 protein at positions corresponding to positions 167 to 183 of the US28 protein encoded by human cytomegalovirus (HCMV) as set forth in SEQ ID NO: 5 may not necessarily be identical to the sequence found in that region of SEQ ID NO: 5, since there may be inter-strain variation in the sequence of ECD3 of US28 amongst different HCMV strains. Accordingly, in this context, the term "corresponding to" refers to the amino acids found in the region that has a corresponding position (that is, in the 2nd extracellular loop, also referred to as the ECD3 region) of the US28 protein encoded by any HCMV strain of interest.

[0302] However, following an analysis of the sequence of the US28 protein from many different clinical and lab strains of HCMV, the applicant has identified the presence of only a single polymorphism within ECD3 of HCMV-encoded US28, thus presenting two alternative ECD3 sequences which are referred to herein as the 4D-variant and the 4N- variant.

[0303] The 4D-variant refers to a sequence variation present in ECD3 of US28, as encoded by a first group of HCMV strains, and appears to be the more common form; around 90% of the sequences of US28 encoded by different HCMV strains, as identified by a BLAST search, show the 4D-variant sequence. The 4D-variant is characterised by comprising the sequence of TKKDNQCMTDYDYLEVS (SEQ ID NO: 7; position 4 of which, as underlined, is D) in ECD3 of US28. Exemplary HCMV strains of the first group, having the 4D-variant of US28 include the Towne, VR1814, TB40 / E, VHL / E, Merlin, JP, Adl69, AF1, BL and DAVIS strains.

[0304] The 4N-variant refers to an alternate sequence variation present in ECD3 of US28, as encoded by a second group of HCMV strains, and appears to be the less common form. The 4D-variant is characterised by comprising the sequence of TKKNNQCMTDYDYLEVS (SEQ ID NO: 6; position 4 of which, as underlined, is N) in ECD3 of US28. Exemplary HCMV strains of the second group, having the 4N-variant of US28 include the Toledo, TR and DB strains. These, and other strains of HCMV showing the 4N-variant sequence in ECD3 of US28 are shown the Table 1.

[0305] The epitope to which the binding molecule of the first aspect of the present invention, such as a HMCV strain agnostic binding molecule of the first aspect of the present invention, has binding specificity is preferably an epitope that is common to, and present within, both the sequence TKKNNQCMTDYDYLEVS (SEQ ID NO: 6, corresponding to the 4N variant of ECD3 of US28) and the sequence of TKKDNQCMTDYDYLEVS (SEQ ID NO: 7, corresponding to the 4D variant of ECD3). To put it another way, the epitope within ECD3 of US28 to which the binding molecule of the first aspect of the present invention has binding specificity, preferably excludes the 4thamino acid residue of each of SEQ ID Nos: 6 and 7, which corresponds to N in the 4N-variant and D in the 4D-variant.

[0306] In the case of linear epitopes, then optionally, epitopes which are common to, and present within, both of the 4N- and 4D-variants, and which exclude the variant 4thamino acid residue, could include all 13 amino acids of the sequence NQCMTDYDYLEVS, or any 12, 11, 10, 9, 8, 7, 6, 5, 4, 3 or fewer amino acids thereof, which may optionally be consecutive amino acids of said sequence.

[0307] In a further option, wherein the epitope is a discontinuous and / or conformational epitope within (preferably entirely within) ECD3 of the US28 protein, then the discontinuous epitope may include any 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 of the 17 amino acids of the 4N variant ECD3 sequence of TKKNNQCMTDYDYLEVS and / or of the 4D variant ECD3 sequence of TKKDNQCMTDYDYLEVS.

[0308] A linear, discontinuous or conformational epitope within ECD3 may exclude one or more of amino acids of ECD3 of US28, which comprises a 1stto through to a 17thposition, within the sequence that corresponds to TKKNNQCMTDYDYLEVS in the 4N variant and TKKDNQCMTDYDYLEVS in the 4D variant.

[0309] In one preferred embodiment, a linear, discontinuous or conformational epitope within ECD3 preferably excludes the 4thposition (N / D) which is known to vary between different HCMV strains.

[0310] Additionally, or alternatively, one or more of the amino acids of ECD3 of US28 may be excluded in the linear, discontinuous or conformational epitope that is bound by binding molecules of the present invention, such as HMCV strain agnostic binding molecules of the first aspect of the present invention.

[0311] For example, a linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 1stposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 2ndposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 3rdposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 5thposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 6thposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 7thposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 8thposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 9thposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 10thposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 11thposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 12thposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 13thposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 14thposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 15thposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 16thposition. A linear, discontinuous or conformational epitope within ECD3 may additionally or alternatively exclude the 17thposition.

[0312] The epitope to which the binding molecule of the first aspect of the present invention, such as a HMCV strain agnostic binding molecule of the first aspect of the present invention, has binding specificity is preferably an epitope that is common to, and present within, greater than 90% of the known clinical and / or lab strains of HCMV (including, at least, all of the HCMV strains disclosed in the present application), for example at least 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or substantially 100% (for example, between 90% and 100%, between 91% and 100%, between 92% and 100%, between 93% and 100%, between 95% and 100%, between 96% and 100%, between 97% and 100%, between 98% and 100%, between 99 and 100%) of the clinical and / or lab strains of HCMV (including, at least, all of the HCMV strains disclosed in the present application).

[0313] In one preferred embodiment, the amino acids in the epitope to which the binding molecule of the first aspect of the present invention has binding specificity may include, or be identical to, the amino acids in the epitope within ECD3 that is bound by any one or more of the US28-13-5G6-1D3, 13-1C10-1C10, 13-1H3-1A10, 13-1C10- 1G9, 14-4E4-1E8 antibodies (commonly abbreviated herein to "1D3", "1C10", "1A10", "1G9" and "1E8", respectively) as described herein (by which is included also an scFv comprising a VH polypeptide sequence having the VH sequence of 1D3, 1C10, 1A10, 1G9 and / or 1E8 as defined by SEQ ID Nos: 12, 104, 122, 68 and 88, respectively, and a VL polypeptide sequence having the VL sequence of 1D3, 1C10, 1A10, 1G9 and / or 1E8 as defined by SEQ ID Nos: 18, 108, 126, 72 and 92, respectively), or vary from said epitope by not more than 5, 4, 3, 2 or 1 amino acids. Binding Properties:

[0314] As reported in the examples of the present application, multiple monoclonal antibodies have been generated against ECD3 of US28, by using the methods described herein, and were shown to bind well and specifically on both genetic variants of the US28 ECD3 peptides and on US28 overexpressing US28-CHO-A1 cells, and certain exemplary antibodies therefrom were further tested and shown to provide excellent binding properties to HCMV Ad 169 infected MRC-5 cells, primary PBMCs from HCMV seropositive individuals, HCMV infected human lung tissue and several types of aggressive human tumors, such as oesophagus, gastric, rectum, liver, lung, pancreas, cervical cancers, malignant pheochromocytoma and locally advanced colon cancer, breast cancer and its metastasis and glioblastoma grade 4. These monoclonal antibodies are exemplary embodiments of binding molecules according to the present invention.

[0315] More specifically, the 1D3, 1C10, 1A10, 1G9 and / or 1E8 antibodies as described herein (by which is included also an scFv comprising a VH polypeptide sequence having the VH sequence of 1D3, 1C10, 1A10, 1G9 and / or 1E8 as defined by SEQ ID Nos: 12, 104, 122, 68 and 88, respectively, and a VL polypeptide sequence having the VL sequence of 1D3, 1C10, 1A10, 1G9 and 1E8 as defined by SEQ ID Nos: 18, 108, 126, 72 and 92, respectively) are preferred exemplary binding molecules according to the present invention, although other binding molecules have been produced and the scope of the first aspect of the present invention is not limited only to 1D3, 1C10, 1A10, 1G9 and / or 1E8, nor only to binding molecules derived therefrom (such as other binding molecules sharing the CDRs of 1D3, 1C10, 1A10, 1G9 and / or 1E8), although these may represent various preferred embodiments. Nevertheless, the 1D3, 1C10, 1A10, 1G9 and / or 1E8 antibodies (by which is included also an scFv comprising a VH polypeptide sequence having the VH sequence of 1D3, 1C10, 1A10, 1G9 and / or 1E8 as defined by SEQ ID Nos: 12, 104, 122, 68 and 88, respectively, and a VL polypeptide sequence having the VL sequence of 1D3, 1C10, 1A10, 1G9 and / or 1E8 as defined by SEQ ID Nos: 18, 108, 126, 72 and 92, respectively) can provide a useful benchmark against which to characterise the binding properties of other binding molecules according to the first aspect of the present invention.

[0316] For example, a binding molecule according to the first aspect of the present invention, having binding specificity to an epitope within extracellular domain 3 (ECD3) of a US28 protein, may demonstrate binding properties that are similar or substantially equivalent to the binding properties of any one, or more, of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8, when tested under the same conditions as 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8. In that context, a binding molecule according to the first aspect of the present invention can be considered to possess "similar or substantially equivalent to the binding properties" to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8, when tested under the same conditions as 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8, respectively, if it displays similar or substantially equivalent binding specificity, similar or substantially equivalent strain agnostic binding properties, and / or similar or substantially equivalent binding affinity, to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8. Such tests may, for example, correspond to any one or more of the tests reported in the present application for assessing the binding properties of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8, for example, any one or more of the tests conducted to determine binding to US28 ECD3 peptides, on US28 overexpressing US28-CHO-A1 cells, HCMV Adl69 infected MRC-5 cells, primary PBMCs from HCMV seropositive individuals, HCMV infected human lung tissue and several types of aggressive human tumors, such as oesophagus, gastric, rectum, liver, lung, pancreas, cervical cancers, malignant pheochromocytoma and locally advanced colon cancer, breast cancer and its metastasis and glioblastoma grade 4.

[0317] A binding molecule according to the first aspect of the present invention may be considered to have similar or substantially equivalent binding specificity to a reference binding molecule of the present invention, for example any one or more of 1D3, 1C10, 1A10, 1G4 and / or 1E8 if, under any one or more tests to determine the ability to bind specifically to US28 ECD3 peptides (compared to a negative control, such as BSA), US28-expressing cells (compared to equivalent cells not expressing US28), HCMV infected cells (compared to equivalent cells without HCMV infection), primary PBMCs from HCMV seropositive individuals (compared to primary PBMCs from HCMV seronegative individuals), HCMV infected human lung tissue (compared to human lung tissue that is not HCMV infected) and / or types of HCMV-infected human tumors, in particular aggressive tumors, such as oesophagus, gastric, rectum, liver, lung, pancreas, cervical cancers, malignant pheochromocytoma and locally advanced colon cancer, breast cancer and its metastasis and glioblastoma grade 4 (compared to equivalent non-cancerous cells that are not infected with HCMV), the binding molecule displays at least 1, 5, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 95, or substantially 100% of the binding specificity of reference binding molecule (e.g. any of 1D3, 1C10, 1A10, 1G4 and / or 1E8). Certain binding molecules according the first aspect of the present invention may have a similar or substantially equivalent binding specificity to the binding specificity of the reference binding molecule (e.g. any one or more of 1D3, 1C10, 1A10, 1G4 and / or 1E8) that is nevertheless lower than the binding specificity of the reference binding molecule (e.g. any one or more of 1D3, 1C10, 1A10, 1G4 and / or 1E8), under any one or more of the foregoing tests; whereas certain other binding molecules according the first aspect of the present invention may have a higher binding specificity than the reference binding molecule (e.g. any one or more of 1D3, 1C10, 1A10, 1G4 and / or 1E8) under any one or more of the foregoing tests.

[0318] A binding molecule according to the first aspect of the present invention may be considered to have similar or substantially equivalent strain agnostic binding properties to a reference binding molecule (e.g. any one or more of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8) if, under any one or more tests to determine the ability to bind specifically to each of 4D- and 4N-variant US28 ECD3 peptides (each compared to a negative control, such as BSA), to each of 4D- and 4N-variant US28-expressing cells (each compared to equivalent cells not expressing US28; and optionally wherein the 4D- and 4N-variants of US28 are, respectively, the US28 sequences encoded by strains TB40 / E and DB of HCMV, and / or further optionally wherein the cells are CHO cells), each of 4D- and 4N-variant US28-encoding HCMV strain infected cells (each compared to equivalent cells without HCMV infection), to primary PBMCs from each of 4D- and 4N- variant US28-encoding HCMV strain seropositive individuals (compared to primary PBMCs from HCMV seronegative individuals), to each of 4D- and 4N-variant US28- encoding HCMV strain infected human lung tissue (compared to human lung tissue that is not HCMV infected) and / or to each of 4D- and 4N-variant US28-encoding HCMV strain types of HCMV-infected human tumors, in particular aggressive tumors, such as oesophagus, gastric, rectum, liver, lung, pancreas, cervical cancers, malignant pheochromocytoma and locally advanced colon cancer, breast cancer and its metastasis and glioblastoma grade 4 (compared to equivalent non-cancerous cells that are not infected with HCMV), the binding molecule displays an equality of binding to the 4D- and 4N-variant that is similar or substantially identical (e.g. ± 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1% or less) to the equality of binding shown by the reference binding molecule (e.g. any one or more of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8).

[0319] A binding molecule according to the first aspect of the present invention may be considered to have similar or substantially equivalent background binding to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8 if, under any one or more tests to determine the background binding to cells that do not express US28 (compared to equivalent cells engineered to express US28), HCMV non-infected cells (compared to equivalent cells with HCMV infection), primary PBMCs from HCMV seronegative individuals (compared to primary PBMCs from HCMV seropositive individuals), HCMV non-infected human lung tissue (compared to human lung tissue that is HCMV infected) and / or types of HCMV non-infected human tumors (compared to equivalent cancerous cells that are infected with HCMV, in particular aggressive tumors, such as oesophagus, gastric, rectum, liver, lung, pancreas, cervical cancers, malignant pheochromocytoma and locally advanced colon cancer, breast cancer and its metastasis and glioblastoma grade 4), the background binding molecule displays at least 1, 5, 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 95, or substantially 100% of the background binding of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8. Certain binding molecules according the first aspect of the present invention may have a similar or substantially equivalent background binding to the background binding of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8 that is nevertheless higher than the background binding of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8, under any one or more of the foregoing tests; whereas certain other binding molecules according the first aspect of the present invention may have lower binding specificity than 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8 under any one or more of the foregoing tests. Alternatively, or additionally, certain binding molecules according the first aspect of the present invention may have lower background binding (e.g. less than 95, 90, 80, 70, 60, 50, 40, 30, 25, 20, 15, 10, 5, 1%, or less than 1%) compared to the background binding of VUNlOO (monovalent VUN100 and / or bivalent VUN 100), under any one or more of the foregoing tests.

[0320] A binding molecule according to the first aspect of the present invention may be considered to have (e.g. ± 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1% or less) substantially equivalent binding affinity to the reference binding molecule (e.g. any one or more of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8) if, under any one or more tests to determine the binding affinity to US28 ECD3 peptides, US28-expressing cells, HCMV infected cells, primary PBMCs from HCMV seropositive individuals, HCMV infected human lung tissue and / or types of HCMV -infected human tumors, in particular aggressive tumors, such as oesophagus, gastric, rectum, liver, lung, pancreas, cervical cancers, malignant pheochromocytoma and locally advanced colon cancer, breast cancer and its metastasis and glioblastoma grade 4, the binding molecule displays a binding affinity that is within, for example, 6, 5, 4, 3, 2 or 1 orders of magnitude of the binding affinity of the reference binding molecule (e.g. any one or more of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8), when expressed as Kd. Certain binding molecules according the first aspect of the present invention may have a lower binding affinity than the reference binding molecule (e.g. any one or more of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8) under any one or more of the foregoing tests; whereas certain other binding molecules according the first aspect of the present invention may have a higher binding affinity than the reference binding molecule (e.g. any one or more of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8) under any one or more of the foregoing tests. It is noted that higher binding affinity is not necessarily always desirable, for example in the context of CARs wherein particularly high binding affinity can be less preferable, and CARs with higher binding specificity generally lead to greater therapeutic benefit than CARs with high binding affinity. A binding molecule according the first aspect of the present invention may be considered to have any one or more of the aforementioned properties. For example, a binding molecule may be considered to have one or more of the following properties (each of which is described in more detail above) :

[0321] 1. similar or substantially equivalent binding specificity to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8;

[0322] 2. similar or substantially equivalent strain agnostic binding properties to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8;

[0323] 3. similar or substantially equivalent background binding to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8;

[0324] 4. lower background binding than the background binding of VUN 100 (monovalent VUN100 and / or bivalent VUN100); and / or

[0325] 5. substantially equivalent binding affinity to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8.

[0326] For example, a binding molecule may have (I) similar or substantially equivalent strain agnostic binding properties to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8; and (II) similar or substantially equivalent background binding to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8. In other embodiments, a binding molecule may have (I) similar or substantially equivalent strain agnostic binding properties to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8; and (II) lower background binding to the background binding of VUN100 (monovalent VUN100 and / or bivalent VUN100). In other embodiments, a binding molecule may have (I) similar or substantially equivalent strain agnostic binding properties to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8; (II) similar or substantially equivalent background binding to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8; and (ill) lower background binding to the background binding of VUN100 (monovalent VUN 100 and / or bivalent VUN100). In other embodiments, a binding molecule may have (I) similar or substantially equivalent strain agnostic binding properties to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8; (II) similar or substantially equivalent background binding to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8; (ill) lower background binding to the background binding of VUN100 (monovalent VUN100 and / or bivalent VUN 100); and (iv) similar or substantially equivalent binding specificity to 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8.

[0327] Further optional assays for charactering the binding properties of binding molecule according to the first aspect of the present invention are discussed below.

[0328] (a) Assay 1:

[0329] As shown in Fig 5 of the present application, an exemplary binding molecule according to the present invention, monoclonal antibody 1D3 that was raised against peptide sequence derived from ECD3 of US28, possesses the ability to bind specifically to peptide sequences derived from ECD3 of US28, compared to a negative control BSA.

[0330] The peptide sequences used in that assay were designated Peptide- 1, having the sequence TKKNNQCMTDYDYLEVS (SEQ ID NO: 6, corresponding to the 4N variant of ECD3) and Peptide-2, having the sequence of TKKDNQCMTDYDYLEVS (SEQ ID NO: 7, corresponding to the 4D variant of ECD3).

[0331] As indicated in Fig 5, the 1D3 antibody shows approximately 27 to 28-fold more specific binding to each of Peptide 1 and 2, compared to BSA control; and the binding levels that are observed of 1D3 to Peptides 1 and 2, both as an absolute level and when normalised against the respective BSA controls, are essentially identical (e.g. less than 5% difference, and likely within the region of experimental error). These results clearly show that 1D3 is both specific for ECD3 of US28, and also strain agnostic.

[0332] In contrast, the binding properties of VUN100 of WO 2019 / 151865 (having the sequence of SEQ ID NO: 60 of the present application) are very different. It is reported to bind to an epitope in the N-terminal region of US28, a region of US28 that possesses high levels of sequence variation between the different HCMV strains. VUN100 is shown to have a relatively low level of binding specificity; a specificity score of 4 was reported by De Groof et a / ., 2019 (supra) in their supporting information, Figure SI. A thereof, and Table 2 of the present application. In addition, VUN 100 shows a considerable difference in binding between the VHL / E, Merlin and TB40 / E strains of HCMV, with binding being particularly reduced in strain TB40 / E (Bl type) at around only half the level of binding observed against the Merlin strain (Figure 8D of WO 2019 / 151865). Further characterisation of VUN 100 is reported in a pre-printed article available online by De Groof et al, 2020 (doi: https: / / doi.org / 10.1101 / 2020.05.12.071860), wherein Fig 2 of the supplementary data gives the results of the % of induced HCMV IE-positive CD 14+ monocytes bound by VUN 100 from four different HCMV seropositive donors, wherein the strain(s) of HCMV within each donor is undetermined. The level of IE positivity induced by VUN 100 binding to these cells varied by up to 14-fold between the different donors. This may be seen as a further indication that the binding ability of VUN 100 varies considerably between cells infected with different strains of HCMV. These disadvantageous properties of VUN 100, in comparison to the binding molecules of the present invention, are further demonstrated in the Examples of the present application, with particular reference to Figure 21A-E, Figure 22 and Figure 23.

[0333] Accordingly, in one embodiment, a binding molecule of the first aspect of the present invention may be characterised as having binding specificity to an epitope within ECD3 of a US28 protein of HCMV if it displays greater binding to Peptide 1 and / or Peptide 2 (preferably both), compared to a negative control (such as BSA), in a binding assay under conditions in which a reference antibody, wherein the reference antibody is 1D3, displays greater binding to Peptide 1 and / or Peptide 2 (preferably both), compared to the same negative control. In another embodiment, the reference binding molecule may be selected from the group consisting of 1C10, 1A10, 1G4, 1G9 and / or 1E8.

[0334] In an additional or alternative embodiment, a binding molecule of the first aspect of the present invention may be characterised as having strain agnostic binding to ECD3 if the respective binding levels that are observed of the binding molecule to each of Peptides 1 and 2 in the aforementioned assay, either as an absolute level and / or when normalised against the binding level observed to the respective negative control (such as BSA) control, are essentially identical, such as within 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1% or less of each other.

[0335] The binding specificity of a binding molecule to an epitope within ECD3 of a US28 protein may, for example, be assessed in an ELISA, such as a methodology as described in the examples of this application, or by a variant of said methodology, such as by coating peptides derived from ECD3 (such as Bio-Peptide 1 and / or Bio-Peptide- 2) on a first set of wells in a microtiter plate and coating a negative control (such as BSA) in a second set of wells in a microtiter plate. The first set of wells may optionally be further subdivided into a first subgroup of the first set of wells that comprises the sequence of a 4N-variant from ECD3 of US28, and a second subgroup of the first set of wells that comprises the sequence of a 4D-variant from ECD3 of US28, to permit the determination of the relative binding specificity to each variant. Following the coating of the first and second sets of wells, a binding molecule of interest may be incubated in each of the first and second sets of wells, optionally with additional testing of a positive control (e.g. any one or more of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8) in respective first and second sets of wells. Following incubation, the wells may be washed and then incubated with an enzyme-conjugated secondary antibody. Following a further wash, a substrate for the conjugated enzyme can be added that undergoes a measurable reaction (e.g. a colour change, the absorbance of) which correlates with the amount of binding for the binding molecule and controls, which gives an indication of binding specificity and / or strain agnostic binding characteristics. Accordingly, in some embodiments, the binding molecule of interest has an absorbance value following ELISA that is indicative of positive binding to one or more (preferably both) of Peptides 1 and / or 2 derived from ECD3 in the first set of wells, compared to the binding to the negative control (such as BSA).

[0336] By "positive binding", we include that the binding molecule of interest can have a higher level of binding to one or more (preferably both) of Peptides 1 and / or 2 compared with the level of binding to a negative control (e.g. BSA), which indicates a higher binding specificity.

[0337] It may, for example, display at least about 2-fold, at least about 5-fold, at least about 10-fold, at least 15-fold, at least 20-fold, at least 25-fold, about 26-fold, about 27-fold, or about 28-fold greater binding to Peptide 1 and / or Peptide 2, compared to a negative control such as BSA under conditions in which antibody 1D3 displays about 27 to 28-fold greater binding to Peptide 1 and / or Peptide 2, compared to a negative control such as BSA. The term "about" as used in this context can include values that are ± 50%, 40%, 30%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% of the stated value (for example, ± 10% of at least about 10-fold refers to the range of from 9-fold to 11-fold). For example, the conditions for such an assay may be selected to be the same as, or equivalent to, the conditions used in the assay used in generating the results shown in Fig 5.

[0338] Alternatively, or additionally, "positive binding" can include that the binding molecule of interest has a similar (e.g. at least ± 50%, 40%, 30%, 20%, 10%, 5% or less), or higher / increased / enhanced / improved, absorbance level compared with a positive control (e.g. 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8).

[0339] (b) Assay 2:

[0340] In a further embodiment, it is preferred that a binding molecule of the present invention binds preferentially to US28-expressing cells (such as CHO cells engineered to express US28) compared to its binding of US28-negative cells (such as wildtype CHO cells), for example, at a 1 : 50 (w / v) dilution, for example as determined by flow cytometry assay. US28-expressing cells are preferably cells that are characterised by displaying cell surface expression of the US28 protein. Surface expression of US28 is typically characterised by the display of the extracellular domains of US28 (ECDs 1, 2, 3 and 4 corresponding, respectively, to positions 1-37, positions 91-101, positions 167- 183 and positions 250-273 of the US28 protein encoded by HCMV strain DB as defined by the sequence of SEQ ID NO: 5, or equivalent sequences of other HCMV strains) on the cell surface.

[0341] Figs 4, 7 and Table 2 of the present application demonstrates that an exemplary binding molecule according to the present invention, monoclonal antibody 1D3 raised against ECD3 of US28, possess the ability to bind specifically to US28-expressing Chinese Hamster Ovary (CHO-US28-A1) cells compared to US28-negative control CHO cells with a specificity score of between around 15 to 21.5 under the binding conditions used. This can be assayed using any suitable technique, for example using FACS analysis, to determine the percentage of total cell count bound. As further reported in Example 1 of the present application, with particular reference to Table 2, although clone US28-13-5G6-1D3 (encoding antibody 1D3) was most fully characterised, other cloned monoclonal antibodies raised against ECD3 of US28 showed excellent binding specificity to CHO-US28-A1 cells, when compared with their binding to US28-negative control CHO cells. As further reported in Example 2, yet further other cloned monoclonal antibodies raised against ECD3 of US28 showed excellent binding specificity to CHO-US28-A1 cells, when compared with their binding to US28-negative control CHO cells, in particular having very low off-target binding (e.g. compared to the much higher levels of off-target binding demonstrated by VUN 100) and in numerous instances also showing higher binding specificity to CHO-US28-A1 cells, when compared with their binding to US28-negative control CHO cells.

[0342] These data clearly demonstrate that, by following the teachings of the present application, there is provided a consistent route to the generation of antibodies raised against ECD3 of US28 that can provide highly specific binding to US28-expressing cells (such as CHO cells) compared to the binding of US28-negative cells (such as US28- negative CHO cells), preferably with low off-target binding levels, a set of characteristics not shared by VUN100.

[0343] In contrast, the binding properties of VUN100 of WO 2019 / 151865 (having the sequence of SEQ ID NO: 60 of the present application) are very different. It is reported to bind to an epitope in the N-terminal region of US28, a region of US28 that possesses high levels of sequence variation between the different HCMV strains. VUN 100 is shown to have a relatively low level of binding specificity: a specificity score of 4 was reported by De Groof et a / ., 2019 (supra) in their supporting information, Figure SI. A thereof (and summarised in Table 2 of the present application) when tested for binding to either US28-expressing HEK293T membranes (HEK+US28) or mock transfected HEK293T membranes. VUN100 is clearly not capable of providing highly specific binding to US28. These disadvantageous properties of VUN100, in comparison to the binding molecules of the present invention, are further demonstrated in the Examples of the present application, with particular reference to Figure 21A-E, Figure 22 and Figure 23.

[0344] Accordingly, in one preferred embodiment, a binding molecule of the first aspect of the present invention will display specific binding, characterised in that its binding specificity for US28-expressing cells (such as CHO-US28-A1 cells, optionally wherein the US28 sequence corresponds to the sequence encoded by strain DB or TB40 / E of HCMV) compared to US28-negative cells (such as CHO cells) is greater than the level of specificity achieved by VUN 100. For example, the level of specificity a binding molecule of the first aspect of the present invention may be greater than the level of specificity achieved by VUN 100 in the same assay by at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 80%, about 90%, about 100% (i.e. about 2-fold), about 150%, about 200% (i.e. about 3-fold), about 300% (i.e. about 4-fold), about 400% (i.e. about 5-fold) or about 500% (i.e. about 6-fold), at the same molar concentration. In this context, the VUN100 comparator may refer to a polypeptide comprising, consisting essentially of, or consisting of, the sequence of any one or more of the sequences of SEQ ID NOs: 60-64. In the context of SEQ ID NOs: 61 and 63, this may include proteins with, or without, the indicated signal peptide sequences.

[0345] In another preferred embodiment, a binding molecule of the first aspect of the present invention will display specific binding, characterised in that its binding specificity for US28-expressing cells (such as CHO-US28-A1 cells, optionally wherein the US28 sequence corresponds to the sequence encoded by strain DB or TB40 / E of HCMV) compared to US28-negative cells (such as CHO cells) is at least about the same level of specificity as any one or more of 1D3, 1C10, 1A10, 1G4 and / or 1E8 in the same assay, at the same molar concentration. The term "about" as used in this context can include values that are ± 80%, 70%, 60%, 50%, 40%, 30%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% of the level of specificity of any one or more of 1D3, 1C10, 1A10, 1G4 and / or 1E8 in the same assay, at the same molar concentration.

[0346] In this context, the 1D3, 1C10, 1A10 and / or 1E8 comparator(s) may refer to an antibody molecule comprising, consisting essentially of, or consisting of: the heavy chain polypeptide sequence of 1D3 as defined by SEQ ID NO: 20 (after removal of the indicated N-terminal leader sequence) and light chain polypeptide sequence of 1D3 as defined by SEQ ID NO: 21 (after removal of the indicated N- terminal leader sequence); the heavy chain polypeptide sequence of 1C10 as defined by SEQ ID NO: 157 (after removal of the indicated N-terminal leader sequence) and light chain polypeptide sequence of 1C10 as defined by SEQ ID NO: 158 (after removal of the indicated N- terminal leader sequence); the heavy chain polypeptide sequence of 1A10 as defined by SEQ ID NO: 161 (after removal of the indicated N-terminal leader sequence) and light chain polypeptide sequence of 1A10 as defined by SEQ ID NO: 162 (after removal of the indicated N- terminal leader sequence); or the heavy chain polypeptide sequence of 1E8 as defined by SEQ ID NO: 153 (after removal of the indicated N-terminal leader sequence) and light chain polypeptide sequence of 1E8 as defined by SEQ ID NO: 154 (after removal of the indicated N- terminal leader sequence).

[0347] Such assays can, for example, be performed to determine the relative binding to US28-expressing cells (such as CHO-US28-A1 cells, optionally wherein the US28 sequence corresponds to the sequence encoded by strain DB or TB40 / E of HCMV) and to US28-negative cells (such as CHO cells), for example, at a molar ratio equivalent to a 1: 50 (w / v) dilution of 1D3, 1C10, 1A10 and / or 1E8, for example as determined by flow cytometry assay.

[0348] (c) Assay 3:

[0349] In a further embodiment, it is preferred that a binding molecule of the first aspect of the present invention can bind preferentially to HCMV-infected cells (such as MRC- 5 cells; cat#CCL-171, RRID: CVCL_0440, American Type Culture Collection (ATCC), Manassas, VA 20110 USA) compared to its binding of equivalent cells without HCMV infection. As shown in Fig 8 of the present application, an exemplary binding molecule according to the present invention, monoclonal antibody 1D3 that was raised against ECD3 of US28, possess the ability to bind to HCMV-infected MRC-5 cells but not to the non-infected (mock) cells, when assayed by using flowcytometry analysis. Under the assay conditions used, 1D3 showed about 5.4% binding to HCMV-infected cells, compared to 0% binding to the non-infected (mock) cells. Further testing, as shown in Fig 9, showed that 1D3 binds specifically (with approximately 16-fold greater specificity) to the surface of the HCMV infected MRC-5 cells as compared with the uninfected cells.

[0350] Accordingly, in a further preferred embodiment, a binding molecule of the first aspect of the present invention will display preferential binding to HCMV-infected cells (such as MRC-5 cells) that is, or is at least, 5-fold, about 6-fold, about 7-fold, about 8- fold, about 9-fold, about 10-fold, about 11-fold, about 12-fold, about 13-fold, about 14-fold, about 15-fold or about 16-fold greater, compared to its binding of equivalent cells without HCMV infection. Such an assay may, for example, be conducted in accordance with the protocol used to generate the results shown in Figs 8 or 9 of the present application, or with an equivalent protocol. The term "about" as used in this context can include values that are ± 50%, 40%, 30%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% of the stated value.

[0351] In a further preferred embodiment, a binding molecule of the first aspect of the present invention will display preferential binding to HCMV-infected cells (such as MRC- 5 cells) compared to its binding of equivalent cells without HCMV infection that is greater than the level of preferential binding achieved by VUN100. For example, the level of preferential binding of a binding molecule of first aspect of the present invention may be greater than the level of preferential binding achieved by VUN 100 in the same assay by at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 80%, about 90%, about 100% (i.e. about 2-fold), about 150%, about 200% (i.e. about 3-fold), about 300% (i.e. about 4-fold), about 400% (i.e. about 5- fold) or about 500% (i.e. about 6-fold), at the same molar concentration. In this context, the VUN100 comparator may refer to a polypeptide comprising, consisting essentially of, or consisting of, the sequence of any one or more of the sequences of SEQ ID NOs: 60-64. In the context of SEQ ID NOs: 61 and 63, this may include proteins with, or without, the indicated signal peptide sequences.

[0352] A binding molecule of the first aspect of the present invention can, for example, display preferential binding to HC MV- infected cells compared to its binding of equivalent cells without HCMV infection that is, or is at least, about 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or substantially 100% of the preferential binding activity of a reference antibody of the present invention (e.g. any one or more of 1D3, 1C10, 1A10, 1G4, 1G9 and / or 1E8) underthe same conditions. The term "about" as used in this context can include values that are ± 50%, 40%, 30%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2% or 1% of the stated value (for example, ± 50% of 10% refers to the range of 5% to 15%). For example, the conditions for such an assay may be selected to be the same as, or equivalent to, the conditions used in the assay used in generating the results shown in Fig 8 or 9.

[0353] Polypeptides

[0354] A binding molecule according to the first aspect of the present invention, comprises, consists essentially of, or consists of one or more polypeptide chains.

[0355] A "polypeptide" is used herein in its broadest sense to refer to a compound of two or more subunit amino acids, amino acid analogs, or other peptidomimetics. The term "polypeptide" thus includes short peptide sequences and also longer polypeptides and proteins. The term "amino acid" as used herein includes the standard twenty genetically-encoded amino acids and their corresponding stereoisomers in the 'D' form (as compared with the natural 'L' form), omega-amino acids other naturally-occurring amino acids, unconventional amino acids (e.g. o,o-disubstituted amino acids, N-alkyl amino acids, etc.) and chemically derivatised amino acids (see below).

[0356] Amino acids herein may be referred to by full name, three letter code or single letter code. When an amino acid is being specifically enumerated, such as "alanine" or "Ala" or "A", the term refers to both L-alanine and D-alanine unless explicitly stated otherwise. Other unconventional amino acids may also be suitable components for polypeptides of the present invention, as long as the desired functional property is retained by the polypeptide. For the peptides shown, each encoded amino acid residue, where appropriate, is represented by a single letter designation, corresponding to the trivial name of the conventional amino acid.

[0357] In one embodiment, the polypeptides as defined herein comprise or consist of L-amino acids.

[0358] It will be appreciated by persons skilled in the art that the polypeptides of the present invention and / or as used as described herein in conjunction with the present invention, may comprise or consist of one or more amino acids which have been modified or derivatised.

[0359] Chemical derivatives of one or more amino acids may be achieved by reaction with a functional side group. Such derivatised molecules include, for example, those molecules in which free amino groups have been derivatised to form amine hydrochlorides, p-toluene sulphonyl groups, carboxybenzoxy groups, t- butyloxycarbonyl groups, chloroacetyl groups or formyl groups. Free carboxyl groups may be derivatised to form salts, methyl and ethyl esters or other types of esters and hydrazides. Free hydroxyl groups may be derivatised to form O-acyl or O-alkyl derivatives. Also included as chemical derivatives are those peptides which contain naturally occurring amino acid derivatives of the twenty standard amino acids. For example: 4-hydroxyproline may be substituted for proline; 5-hydroxylysine may be substituted for lysine; 3-methylhistidine may be substituted for histidine; homoserine may be substituted for serine, and ornithine for lysine. Derivatives also include peptides containing one or more additions or deletions as long as the requisite activity is maintained. Other included modifications are amidation, amino terminal acylation (e.g. acetylation or thioglycolic acid amidation), terminal carboxylamidation (e.g. with ammonia or methylamine), and the like terminal modifications.

[0360] It will be further appreciated by persons skilled in the art that peptidomimetic compounds may also be useful. The term 'peptidomimetic' refers to a compound that mimics the conformation and desirable features of a particular peptide as a therapeutic agent.

[0361] For example, the said polypeptide includes not only molecules in which amino acid residues are joined by peptide (-CO-NH-) linkages but also molecules in which the peptide bond is reversed. Such retro-inverso peptidomimetics may be made using methods known in the art, for example such as those described in Meziere et a / . (1997, J. Immunol., 159(7) : 3230-3237). This approach involves making pseudo-peptides containing changes involving the backbone, and not the orientation of side chains. Retro-inverse peptides, which contain NH-CO bonds instead of CO-NH peptide bonds, are much more resistant to proteolysis. Alternatively, the said polypeptide may be a peptidomimetic compound wherein one or more of the amino acid residues are linked by a -y(CH2NH)- bond in place of the conventional amide linkage.

[0362] In a further alternative, the peptide bond may be dispensed with altogether provided that an appropriate linker moiety which retains the spacing between the carbon atoms of the amino acid residues is used; it may be advantageous for the linker moiety to have substantially the same charge distribution and substantially the same planarity as a peptide bond. It will also be appreciated that the said polypeptide may conveniently be blocked at its N- or C-terminus so as to help reduce susceptibility to exo-proteolytic digestion.

[0363] A variety of un-coded or modified amino acids such as D-amino acids and N- methyl amino acids have also been used to modify mammalian peptides. In addition, a presumed bioactive conformation may be stabilised by a covalent modification, such as cyclisation or by incorporation of lactam or other types of bridges, for example see Veber et al. (1978, Proc. Natl. Acad. Sci. USA, 75:2636) and Thursell et at. (1983, Biochem. Biophys. Res. Comm. I l l : 166), which are incorporated herein by reference.

[0364] Binding Molecule Structures:

[0365] A "binding molecule" in accordance with the first aspect of the present invention typically comprises, consists essentially of, or consists of, one or more polypeptides.

[0366] In some embodiments, a binding molecule (or a portion thereof) may be formed through the combination of multiple polypeptides. For example, a VH polypeptide may be combined with a VL polypeptide, therein forming a Fab fragment. The combination of polypeptides may be a VH polypeptide from one exemplary ECD3-binding molecule combined with a VL polypeptide of the same exemplary ECD3-binding molecule, or with a VL polypeptide of a different exemplary ECD3-binding molecule.

[0367] In some preferred embodiments, the binding molecule is selected from the group consisting of an antibody and a chimeric antigen receptor (CAR).

[0368] As discussed above, and further elucidated in Table 2 of Example 1, and also in Example 2, the present application describes numerous antibodies, raised against ECD3 of US28 in accordance with the methods described herein, with highly beneficial binding properties to US28, including the antibodies 13-5G6-1D3 (generally abbreviated herein as "1D3"), 13-5C6-1B5, 14-1H3-1A6, 13-1C10-1C10 (generally abbreviated herein as "1C10"), 14-1H3-1A10 (generally abbreviated herein as "1A10"), 14-2C2-1G4 (generally abbreviated herein as "1G4"), 13-1C10-1G9 (generally abbreviated herein as"lG9") and 14-4E4-1E8 (generally abbreviated herein as "1E8").

[0369] The present application also provides a method of obtaining further antibodies having binding specificity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of HCMV, in accordance with the thirty-fourth aspect of the present invention, as discussed further in Section N of this application. For example, the method may comprise:

[0370] (a) providing one or more peptides corresponding an amino acid sequence present in ECD3 of the US28 protein, such as one or both that peptides comprise, consist essentially of, or consist of, the polypeptide sequence TKKDNQCMTDYDYLEVS (SEQ ID NO: 7) and / or TKKNNQCMTDYDYLEVS (SEQ ID NO: 6), and / or an immunogenic fragment of either or both;

[0371] (b) providing one or more candidate antibodies (which have optionally been raised in response to one or both of said peptides); and

[0372] (c) determining the binding specificity and / or binding affinity of one or more candidate antibodies to the one or both of said peptides.

[0373] In some preferred embodiments, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 1D3 as defined by SEQ ID NOs: 8, 9, 10, 14, 15 and 16, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1D3, and optionally wherein the binding molecule is selected from an antibody and a CAR.

[0374] The sequences, and identities of the CDR sequences of antibody 1D3 as defined by SEQ ID NOs: 8, 9, 10, 14, 15 and 16 are as follows:

[0375] In another embodiment, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 1C10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83, 118, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1C10, and optionally wherein the binding molecule is selected from an antibody and a CAR.

[0376] The sequences, and identities of the CDR sequences of antibody 1C10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83, 118 are as follows:

[0377] In another embodiment, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 1A10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83, 118, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1A10, and optionally wherein the binding molecule is selected from an antibody and a CAR.

[0378] The sequences, and identities of the CDR sequences of antibody 1A10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83, 118 are as follows:

[0379] In another embodiment, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 1G9 as defined by SEQ ID NOs: 76, 77, 78, 82, 83 and 84, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1G9, and optionally wherein the binding molecule is selected from an antibody and a CAR.

[0380] The sequences, and identities of the CDR sequences of antibody 1G9 as defined by SEQ ID NOs: 76, 77, 78, 82, 83 and 84 are as follows:

[0381] In another embodiment, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody 1E8 as defined by SEQ ID NOs: 76, 95, 96, 82, 99 and 100, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1E8, and optionally wherein the binding molecule is selected from an antibody and a CAR.

[0382] The sequences, and identities of the CDR sequences of antibody 1E8 as defined by SEQ ID NOs: 76, 95, 96, 82, 99 and 100 are as follows:

[0383] In other embodiments, the binding molecule of the first aspect of the present invention comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences (and / or a functional variant of any one or more of said CDR sequences) of any other antibody having binding specificity (and preferably a strain agnostic binding specificity) to the ECD3 of US28, such as either of antibodies 13-5C6-1B5 and 14-1H3- 1A6 as described herein, and / or any antibody that is obtained by the method of obtaining further antibodies having binding specificity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of HCMV), as described above.

[0384] It is appreciated that molecules containing three or fewer CDR regions (in some cases just a single CDR or a part thereof) can be capable of retaining the antigenbinding activity of the antibody from which the CDR(s) are derived. For example, Gao et al (1994, J Biol Chem 269: 32389-93) describe a whole VL chain (including all three CDRs) having high affinity for its substrate.

[0385] Molecules containing two CDR regions are described, for example, by Vaughan & Sollazzo (2001, Combinatorial Chemistry & High Throughput Screening , 4: 417-430). On page 418 (right column - 3 Our Strategy for Design) a minibody including only the Hl and H2 CDR hypervariable regions interspersed within framework regions is described. The minibody is described as being capable of binding to a target. Pessi et al (1993, Nature, 362: 367-9) and Bianchi et al (1994, J. Mol. Biol., 236: 649-59) are referenced by Vaughan & Sollazzo and describe the Hl and H2 minibody and its properties in more detail. Qiu et al (2007, Nature Biotechnology, 25:921-9) demonstrate that a molecule consisting of two linked CDRs are capable of binding antigen (abstract and page 926, right-hand column). Quiocho (1993, Nature, 362: 293-4) provides a summary of the Pessi et al. "minibody" technology. Ladner (2007, Nature Biotechnology, 25:875-7) reviews the Qiu et al. article and comments that molecules containing two CDRs are capable of retaining antigen-binding activity (page 875, right-hand column).

[0386] Molecules containing a single CDR region are described, for example, by Laune et al (1997, JBC, 272: 30937-44) who demonstrate that a range of hexapeptides derived from a CDR display antigen-binding activity (abstract) and note that synthetic peptides of a complete, single, CDR display strong binding activity (page 30942, righthand column). Monnet et al (1999, JBC, 274: 3789-96) show that a range of 12-mer peptides and associated framework regions have antigen-binding activity (abstract) and comment that a CDR3-like peptide alone is capable of binding antigen (page 3785, left-hand column). Heap et al (2005, J. Gen. Virol., 86: 1791-1800) report that a "micro-antibody" (a molecule containing a single CDR) is capable of binding antigen (abstract and page 1791, left-hand column) and shows that a cyclic peptide from an anti-HIV antibody has antigen-binding activity and function. Nicaise et al (2004, Protein Science, 13: 1882-91) show that a single CDR can confer antigen-binding activity and affinity for its lysozyme antigen.

[0387] Where functional variants of particular CDR sequences of an antibody are mentioned, it will be appreciated that one or more of the CDRs in the antibody as defined may be varied. Thus, where the antibody is defined as comprising light chain or heavy chain CDRs (e.g. CDRs 1-3), each having a particular sequence, up to one, two, or three of those particular sequences may be varied. Similarly, where the antibody is defined as comprising light chain and heavy chain CDRs (e.g. six CDRs), each having a particular sequence, up to one, two, three, four, five, or all six of those particular sequences may be varied. The functional variants are typically conservative amino acid substitutions as described further below and / or can include amino acid deletions and / or insertions.

[0388] For example, the VH-CDR1 sequence of 1D3 is an 8-amino acid sequence GFTFTDYY (SEQ ID NO: 8). A functional variant thereof may include one or more (e.g. 1, 2, 3, 4, 5, 6, 7 or 8) conservative amino acid substitutions as described further below and / or can include one or more (e.g. 1, 2, 3, 4, or 5) amino acid deletions and / or one or more amino acid insertions. The VH-CDR1 sequences of 1C10, 1A10, 1G9 and 1E8 are 5-amino acid sequences (SEQ ID NOs: 112, 112, 138, 76 and 76, respectively). Functional variants thereof may include one or more (e.g. 1, 2, 3, 4 or 5) conservative amino acid substitutions as described further below and / or can include one or more (e.g. 1 or 2) amino acid deletions and / or one or more amino acid insertions.

[0389] The VH-CDR2 sequence of 1D3 is a 10-amino acid sequence IRSKANGYTT (SEQ ID NO: 9). A functional variant thereof may include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid substitutions as described further below and / or can include one or more (e.g. 1, 2, 3, 4, 5, 6 or 7) amino acid deletions and / or one or more amino acid insertions. The VH-CDR2 sequences of 1C10, 1A10, 1G9 and 1E8 are 16-amino acid sequences (SEQ ID NOs: 113, 113, 77 and 95, respectively). Functional variants thereof may include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16) conservative amino acid substitutions as described further below and / or can include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13) amino acid deletions and / or one or more amino acid insertions.

[0390] The VH-CDR3 sequence of 1D3 is a 12-amino acid sequence ARDERRTAWLAY (SEQ ID NO: 10). A functional variant thereof may include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12) conservative amino acid substitutions as described further below and / or can include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8 or 9) amino acid deletions and / or one or more amino acid insertions. The VH-CDR3 sequence of 1G9 is a 14-amino acid sequence (SEQ ID NO: 78). A functional variant thereof may include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14) conservative amino acid substitutions as described further below and / or can include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11) amino acid deletions and / or one or more amino acid insertions. The VH-CDR3 sequences of 1C10, 1A10 and 1E8 are 13-amino acid sequences (SEQ ID NOs: 114, 114 and 96, respectively). Functional variants thereof may include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13) conservative amino acid substitutions as described further below and / or can include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) amino acid deletions and / or one or more amino acid insertions.

[0391] The VL-CDR1 sequence of 1D3 is an 11-amino acid sequence QSIVHSNGNTY (SEQ ID NO: 14). A functional variant thereof may include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid substitutions as described further below and / or can include one or more (e.g. 1, 2, 3, 4, 5, 6, 7 or 8) amino acid deletions and / or one or more amino acid insertions. The VL-CDR1 sequences of 1C10, 1A10, 1G9 and 1E8 are 10-amino acid sequences (SEQ ID NOs: 117, 117, 82 and 82, respectively). Functional variants thereof may include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid substitutions as described further below and / or can include one or more (e.g. 1, 2, 3, 4, 5, 6 or 7) amino acid deletions and / or one or more amino acid insertions.

[0392] The VL-CDR2 sequence of 1D3 is a 3-amino acid sequence KVS (SEQ ID NO: 15). A functional variant thereof may include one or more (e.g. 1, 2, 3) conservative amino acid substitutions as described further below and / or can include one or more amino acid deletions and / or one or more amino acid insertions. The VL-CDR2 sequences of 1C10, 1A10, 1G9 and 1E8 are 7-amino acid sequences (SEQ ID NOs: 83, 83, 145, 83 and 99, respectively). Functional variants thereof may include one or more (e.g. 1, 2, 3, 4, 5, 6 or 7) conservative amino acid substitutions as described further below and / or can include one or more (e.g. 1, 2, 3 or 4) amino acid deletions and / or one or more amino acid insertions.

[0393] The VL-CDR3 sequence of 1D3 is a 10-amino acid sequence FQGSHVPTWT (SEQ ID NO: 16). A functional variant thereof may include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid substitutions as described further below and / or can include one or more (e.g. 1, 2, 3, 4, 5, 6 or 7) amino acid deletions and / or one or more amino acid insertions. The VL-CDR3 sequences of 1C10, 1A10, 1G9 and 1E8 are 10-amino acid sequences (SEQ ID NOs: 118, 118, 84 and 100, respectively). A functional variant thereof may include one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) conservative amino acid substitutions as described further below and / or can include one or more (e.g. 1, 2, 3, 4, 5, 6 or 7) amino acid deletions and / or one or more amino acid insertions. Functional variants of one or more of said CDRs can be created by any of the various processes well known in the art. For example, the binding properties of a binding molecule that comprises 1, 2, 3, 4, 5 or 6 CDRs can be developed by a maturation process. For example, the binding affinity provided the CDRs of a binding molecule can be modified (increased, or reduced) by maturation steps; and / or the binding specificity provided the CDRs of a binding molecule can be modified (in general, increased) by maturation steps. It is to be understood that high, or increased, levels of binding affinity to US28 may not always be desirable, in particular if this comes at the expense of unacceptably high levels of binding affinity to off-targets, such as healthy human cells. Binding molecules, including but not limited to CARs and CAR- expressing cells, for example, may benefit from lower levels of binding affinity, but will generally always benefit from optimised levels of binding specificity.

[0394] In one example of a maturation process, a monovalent display phagemid system may be used to modify the avidity effects during antigen-binding screening. Two alternative or combined methods, untargeted mutagenesis and oligonucleotide- directed mutagenesis, can be employed to construct random or defined sublibraries to introduce a large number of mutants of the original binding molecule. The binding molecules that bind to US28-expressing cells, and / or to HCMV-infected cells, with the desired modified properties (such as increased specificity and / or increased or decreased affinity) are then selected by modifying the screening conditions, such as in an assay for stringency.

[0395] Binding molecule of the first aspect of the present invention that comprise functional variants of any of the one, two, three, four, five or all six of the CDR sequences of antibody:

[0396] (a) 1D3 as defined by SEQ ID NOs: 8, 9, 10, 14, 15 and 16, respectively;

[0397] (b) 1C10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively;

[0398] (c) 1A10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively;

[0399] (d) 1G9 as defined by SEQ ID NOs: 76, 77, 78, 82, 83 and 84, respectively; and / or

[0400] (e) 1E8 as defined by SEQ ID NOs: 76, 95, 96, 82, 99 and 100, respectively; and preferably possess one or more binding properties that are similar or substantially equivalent to the binding properties of 1D3, 1C10, 1A10, 1G9 and / or 1E8, when tested under the same conditions as 1D3, 1C10, 1A10, 1G9 and / or 1E8, respectively, as further described above.

[0401] In one embodiment, the binding molecule of the first aspect of the present invention may comprise: (a) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable heavy chain (VH) of antibody: i. 1D3, as defined by SEQ ID NOs: 8, 9, and 10, respectively (and / or a functional variant of any one, two or three of said CDR sequences);

[0402] II. 1C10, as defined by SEQ ID NOs: 112, 113, and 114, respectively (and / or a functional variant of any one, two or three of said CDR sequences); ill. 1A10, as defined by SEQ ID NOs: 112, 113, and 114, respectively (and / or a functional variant of any one, two or three of said CDR sequences); iv. 1G9, as defined by SEQ ID NOs: 76, 77, and 78, respectively (and / or a functional variant of any one, two or three of said CDR sequences); and / or v. 1E8, as defined by SEQ ID NOs: 76, 95, and 96, respectively (and / or a functional variant of any one, two or three of said CDR sequences); and / or

[0403] (b) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable light chain (VL) of antibody: i. 1D3, as defined by SEQ ID NOs: 14, 15, and 16, respectively (and / or a functional variant of any one, two or three of said CDR sequences);

[0404] II. 1C10, as defined by SEQ ID NOs: 117, 83, and 118, respectively (and / or a functional variant of any one, two or three of said CDR sequences); ill. 1A10, as defined by SEQ ID NOs: 117, 83, and 118, respectively (and / or a functional variant of any one, two or three of said CDR sequences); iv. 1G9, as defined by SEQ ID NOs: 82, 83, and 84, respectively (and / or a functional variant of any one, two or three of said CDR sequences); and / or v. 1E8, as defined by SEQ ID NOs: 82, 99, and 100, respectively (and / or a functional variant of any one, two or three of said CDR sequences).

[0405] Additionally, or alternatively, a binding molecule of the first aspect of the present invention may comprise:

[0406] (a) at least one variable heavy chain (VH) polypeptide that comprises CDR 1, 2, and 3 sequences having the sequences of: i. SEQ ID NOs: 8, 9, and 10, respectively (and / or a functional variant of any one, two or three of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 12 (or a functional variant thereof);

[0407] II. SEQ ID NOs: 112, 113, and 114, respectively (and / or a functional variant of any one, two or three of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 104 (or a functional variant thereof); ill. SEQ ID NOs: 112, 113, and 114, respectively (and / or a functional variant of any one, two or three of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 122 (or a functional variant thereof); iv. SEQ ID NOs: 76, 77, and 78, respectively (and / or a functional variant of any one, two or three of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 68 (or a functional variant thereof); and / or v. SEQ ID NOs: 76, 95, and 96, respectively (and / or a functional variant of any one, two or three of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 88 (or a functional variant thereof); and / or

[0408] (b) at least one variable light chain (VL) polypeptide that comprises CDR 1, 2 and 3 sequences having the sequences of: i. SEQ ID NOs: 14, 15, and 16, respectively (and / or a functional variant of any one, two or three of said CDR sequences), and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 18;

[0409] II. SEQ ID NOs: 117, 83, and 118, respectively (and / or a functional variant of any one, two or three of said CDR sequences), and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 108; ill. SEQ ID NOs: 117, 83, and 118, respectively (and / or a functional variant of any one, two or three of said CDR sequences), and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 126; iv. SEQ ID NOs: 82, 83, and 84, respectively (and / or a functional variant of any one, two or three of said CDR sequences), and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 72; and / or v. SEQ ID NOs: 82, 99, and 100, respectively (and / or a functional variant of any one, two or three of said CDR sequences), and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 92.

[0410] The sequences, and identities of the variable heavy chain (VH) variable light chain (VL) polypeptide sequence of antibody 1D3 as defined by SEQ ID NOs: 12 and

[0411] 18, respectively are as follows:

[0412] The sequences, and identities of the variable heavy chain (VH) variable light chain (VL) polypeptide sequence of antibody 1C10 as defined by SEQ ID NOs: 104 and

[0413] 108, respectively are as follows:

[0414] The sequences, and identities of the variable heavy chain (VH) variable light chain (VL) polypeptide sequence of antibody 1A10 as defined by SEQ ID NOs: 122 and 126, respectively are as follows:

[0415] The sequences, and identities of the variable heavy chain (VH) variable light chain (VL) polypeptide sequence of antibody 1G9 as defined by SEQ ID NOs: 68 and

[0416] 72, respectively are as follows:

[0417] The sequences, and identities of the variable heavy chain (VH) variable light chain (VL) polypeptide sequence of antibody 1E8 as defined by SEQ ID NOs: 88 and

[0418] 92, respectively are as follows:

[0419] In other embodiments, the binding molecule of the first aspect of the present invention comprises (a) at least one variable heavy chain (VH) polypeptide that comprises CDR 1, 2, and 3 sequences of the VH chain of another antibody having binding specificity (and preferably a strain agnostic binding specificity) to the ECD3 of US28 (and / or a functional variant of any one, two or three of said CDR sequences), and / or (b) at least one variable light chain (VL) polypeptide that comprises CDR 1, 2 and 3 sequences of the VL chain of the same other antibody having binding specificity (and preferably a strain agnostic binding specificity) to the ECD3 of US28 (and / or a functional variant of any one, two or three of said CDR sequences). Said other antibody can, for example, be either of antibodies 13-5C6-1B5 and 14-1H3-1A6 as described herein, and / or another antibody that is obtained by the method of obtaining further antibodies having binding specificity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of HCMV, as described above. When the antibody is defined as having a light chain variable region comprising a particular amino acid sequence and a heavy chain variable region having a particular amino acid sequence, it will be appreciated that up to one or two of those sequences may be varied as defined. The variation may be solely within the non-CDR regions of the sequences, solely within the CDR regions of the sequences, or within both the non- CDR and CDR regions of the sequences. Typically, the variation is outside of the CDR regions, and so the variants of the light chain variable regions and heavy chain variable regions, may comprise any of the particular CDR sequences defined herein.

[0420] Where the antibody comprises a variant of a heavy chain variable region and / or a light chain variable region as defined herein (e.g. a VH selected from SEQ ID NOs: 12, 104, 122, 130, 68 and 88 and / or a VL selected from SEQ ID NOs: 18, 108, 126, 134, 72 and 92), the variant may have at least 1, 2, 3, 4, or 5 amino acid substitutions. Typically, the variants do not have more than 30, 20, or 10 amino acid substitutions. Hence, the variants may have at least 1, 2, 3, 4 or 5 amino acid substitutions but not more than 30, 20 or 10 amino acid substitutions.

[0421] Where the antibody comprises a variant of a heavy chain variable region and / or a light chain variable region as defined herein (e.g. a VH selected from SEQ ID NO: 12, 104, 122, 130, 68 and 88 and / or a VL selected from SEQ ID NO: 18, 108, 126, 134, 72 and 92), the variant generally has at least 70% sequence identity to the defined amino acid sequence, for example at least 75%, 80%, 85%, 90% or 95% sequence identity, and more generally has 95-99% sequence identity to the defined amino acid sequence (e.g. 96%, 97%, 98% or 99% sequence identity). The level of variation may be applicable solely to the non-CDR region. For example, a variant of a heavy chain variable region and / or a light chain variable region as defined herein (e.g. a VH selected from SEQ ID NO: 12, 104, 122, 130, 68 and 88 and / or a VL selected from SEQ ID NO: 18, 108, 126, 134, 72 and 92), may have at least 70% identity (e.g. 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity) to the defined amino acid sequence but comprise one or more (such as all) identical CDRs as defined herein.

[0422] The percent sequence identity between two polypeptides may be determined using suitable computer programs, for example the GAP program of the University of Wisconsin Genetic Computing Group and it will be appreciated that percent identity is calculated in relation to polypeptides whose sequence has been aligned optimally. The alignment may alternatively be carried out using the Clustal W program (Thompson et al., (1994) Nucleic Acids Res 22, 4673-80). The parameters used may be as follows: Fast pairwise alignment parameters: K-tuple(word) size; 1, window size; 5, gap penalty; 3, number of top diagonals; 5. Scoring method : x percent. Multiple alignment parameters: gap open penalty; 10, gap extension penalty; 0.05. Scoring matrix: BLOSUM. Binding molecule of the first aspect of the present invention that comprise functional variants of the variable heavy chain (VH) and / or variable light chain (VL) polypeptide sequence of antibody 1D3, 1C10, 1A10, 1G9 and / or 1E8 as defined by SEQ ID NOs: 12, 104, 122, 68 and 88 for the VH and 18, 108, 126, 72 and 92 for the VL, respectively, preferably possess one or more binding properties that are similar or substantially equivalent to the binding properties of 1D3, 1C10, 1A10, 1G9 and / or 1E8, when tested under the same conditions as 1D3, 1C10, 1A10, 1G9 and / or 1E8, respectively, as further described above.

[0423] Typically, it is preferred that the amino acid substitutions of the variants disclosed herein are conservative amino acid substitutions, for example where an amino acid residue is replaced with an amino acid residue having a similar side chain. Conservative amino acid substitutions are well known in the art and include (original residue substitution) Ala (A) Vai, Gly or Pro; Arg (R) Lys or His; Asn (N) Gin;

[0424] Asp Cys (C) Ser; Gin Glu (G) Asp; Gly (G) Ala; His (H)

[0425] Arg; lie (I) Leu; Leu (L) lie, Vai or Met; Lys (K) Arg; Met (M) Leu; Phe (F) Tyr; Pro (P) Ala; Ser ( Thr or Cys; Thr (T) Ser; Trp Tyr (Y) Leu or Ala.

[0426] Antibodies:

[0427] In one non-limiting, but preferred embodiment, a binding molecule according to the first aspect of the present invention may be, of comprise, an antibody.

[0428] The term "antibody" as used herein optionally includes antigen-binding fragments of said antibody. The term "chimeric antigen receptor (CAR)" as used herein optionally includes antigen-binding fragments of said CAR.

[0429] By "antibody or antigen-binding fragment thereof" we include substantially intact antibody molecules, as well as chimeric antibodies, humanised antibodies, isolated human antibodies, single chain antibodies, monospecific antibodies, bispecific antibodies, antibody heavy chains, antibody light chains, homodimers and heterodimers of antibody heavy and / or light chains, and antigen-binding fragments and derivatives of the same. Suitable antigen-binding fragments and derivatives include Fv fragments (e.g. single chain Fv and disulphide-bonded Fv), Fab-like fragments (e.g. Fab fragments, Fab' fragments and F(ab)2 fragments), single variable domains (e.g. VH and VL domains) and single domain antibodies (dAbs, including single and dual formats [i.e. dAb-linker-dAb], and nanobodies). The potential advantages of using antibody fragments, rather than whole antibodies, are several-fold. The smaller size of the fragments may lead to improved pharmacological properties, such as better penetration of solid tissue. Moreover, antigen-binding fragments such as Fab, Fv, ScFv and dAb antibody fragments can be expressed in and secreted from recombinant sources (e.g. CHO cells, E. coli, etc.) thus allowing the facile production of large amounts of the said fragments.

[0430] By "an antibody" or "an antigen-binding fragment thereof" we include substantially intact antibody molecules, as well as chimeric antibodies, humanised antibodies, and isolated human antibodies.

[0431] An "an antibody", including an "antigen-binding fragment thereof", a CAR or antigen-binding fragment thereof, and / or other binding molecule in accordance with the first aspect of the present invention may, for example: i. have a valency of "n", wherein the n is an integer of one or more, and so, for example, may be monovalent, bivalent, trivalent or multivalent; and / or

[0432] II. have binding specificity to one antigen, or to two or more different antigens, for example it may have the ability to bind specifically to "x" different antigens, wherein x is an integer of one, or two or more, subject to the requirement that at least one antigen is the ECD3 of the US28 protein; for example, it may have mono-specific, bi-specific, tri-specific or multi-specific binding specificity.

[0433] Exemplary forms of antibody or antigen-binding fragments thereof can include any one or more of single chain antibodies, bispecific antibodies, antibody heavy chains, antibody light chains, homodimers and heterodimers of antibody heavy and / or light chains, and antigen-binding fragments and derivatives of the same. Suitable antigen-binding fragments and derivatives include Fv fragments (e.g. single chain Fv and disulphide-bonded Fv), Fab-like fragments (e.g. Fab fragments, Fab' fragments and F(ab)2 fragments), single variable domains (e.g. VH and VL domains) and single domain antibodies (dAbs, including single and dual formats [i.e. dAb-linker-dAb], and nanobodies). The potential advantages of using antibody fragments, rather than whole antibodies, are several-fold. The smaller size of the fragments may lead to improved pharmacological properties, such as better penetration of solid tissue. Moreover, antigen-binding fragments such as Fab, Fv, single chain Fv (ScFv) and dAb antibody fragments can be expressed in and secreted from recombinant host cells, such as E. coli, thus allowing the facile production of large amounts of the said fragments.

[0434] The term "bi-specific" as used herein means the polypeptide is capable of specifically binding at least two target entities. Each of these target entities may be to epitopes derived from the same protein (e.g. binding to a first epitope and second epitope of the same protein, e.g. US28). Alternatively, the target entities may be to epitopes derived from different proteins (e.g. binding to a first epitope of a first protein, and a second epitope of a second, different target, such as a different protein).

[0435] By "ScFv molecules" we mean molecules wherein the VH and VL partner domains are linked via a flexible oligopeptide. Engineered antibodies, such as ScFv antibodies, can be made using the techniques and approaches long known in the art. The advantages of using antibody fragments, rather than whole antibodies, are several-fold. The smaller size of the fragments may lead to improved pharmacological properties, such as better penetration to the target site. Effector functions of whole antibodies, such as complement binding, are removed. Fab, Fv, ScFv and dAb antibody fragments can all be expressed in and secreted from E. coll, thus allowing the facile production of large amounts of the fragments. Whole antibodies, and F(ab')2 fragments are "bivalent". By "bivalent" we mean that the antibodies and F(ab')2 fragments have two antigen combining sites. In contrast, Fab, Fv, ScFv and dAb fragments are monovalent, having only one antigen combining site.

[0436] The phrases "an antibody" or "an antigen-binding fragment thereof" is also intended to encompass antibody mimics (for example, non-antibody scaffold structures that have a high degree of stability yet allow variability to be introduced at certain positions). Those skilled in the art of biochemistry will be familiar with many such molecules, as discussed in Gebauer & Skerra, 2009. Exemplary antibody mimics include: affibodies (also called Trinectins; Nygren, 2008, FEBS J, 275, 2668-2676); CTLDs (also called Tetranectins; Innovations Pharmac. Technol. (2006), 27-30); adnectins (also called monobodies; Meth. Mol. Biol., 352 (2007), 95-109); anticalins (Drug Discovery Today (2005), 10, 23-33); DARPins (ankyrins; Nat. Biotechnol. (2004), 22, 575-582); avimers (Nat. Biotechnol. (2005), 23, 1556-1561); microbodies (FEBS J, (2007), 274, 86-95); peptide aptamers (Expert. Opin. Biol. Ther. (2005), 5, 783-797); Kunitz domains (J. Pharmacol. Exp. Ther. (2006) 318, 803-809); affilins (Trends. Biotechnol. (2005), 23, 514-522); affimers (Avacta Life Sciences, Wetherby, UK).

[0437] Persons skilled in the art will further appreciate that the invention also encompasses modified versions of antibodies and antigen-binding fragments thereof, whether existing now or in the future, e.g. modified by the covalent attachment of polyethylene glycol or another suitable polymer (see below) and / or covalently or non- covalently attached (e.g. by presentation as a fusion protein) to other polypeptide sequences.

[0438] Methods of generating antibodies and antibody fragments are well known in the art. For example, antibodies may be generated via any one of several methods which employ induction of in vivo production of antibody molecules, screening of immunoglobulin libraries (Orland! et al., 1989; Winter et al., 1991) or generation of monoclonal antibody molecules by cell lines in culture. These include, but are not limited to, the hybridoma technique, the human B-cell hybridoma technique, and the Epstein-Barr virus (EBV)-hybridoma technique (Kohler at al., 1975. Nature 256:4950497; Kozbor et a / ., 1985. J. Immund. Methods 81 :31-42; Cote et al., 1983. Proc. Natl. Acad. Sci. USA 80:2026-2030; Cole et a / ., 1984. Mol. Cell. Biol. 62: 109- 120)

[0439] Suitable methods for the production of monoclonal antibodies are also disclosed in "Monoclonal Antibodies: A manual of techniques", H Zola (CRC Press, 1988) and in "Monoclonal Hybridoma Antibodies: Techniques and Applications", J G R Hurrell (CRC Press, 1982).

[0440] The term "monoclonal antibody" as used herein includes reference to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts. Monoclonal antibodies are highly specific, being directed against a single antigenic site. Furthermore, in contrast to conventional (polyclonal) antibody preparations which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen. In addition to their specificity, the monoclonal antibodies are advantageous in that they are synthesised by a hybridoma culture, uncontaminated by other immunoglobulins. The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies, and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler et at, Nature, 256: 495 (1975), or may be made by recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567). The "monoclonal antibodies" may also be isolated from phage antibody libraries using the techniques described in Clackson et at, Nature, 352:624-628 (1991) and Marks et at, J. Mol. Biol., 222: 581-597 (1991), for example.

[0441] Likewise, antibody fragments can be obtained using methods well known in the art (see, for example, Harlow & Lane, 1988, "Antibodies: A Laboratory Manual", Cold Spring Harbor Laboratory, New York). For example, antibody fragments according to the present invention can be prepared by proteolytic hydrolysis of the antibody or by expression in E. coli or mammalian cells (e.g. Chinese hamster ovary [CHO] cell culture or other protein expression systems) of DNA encoding the fragment. Alternatively, antibody fragments can be obtained by pepsin or papain digestion of whole antibodies by conventional methods.

[0442] It will be appreciated by persons skilled in the art that for human therapy and / or diagnostics, human or humanised antibodies are preferably used. Humanised forms of non-human (e.g. murine) antibodies are genetically engineered chimeric antibodies or antibody fragments (e.g. antigen-binding fragments) having preferably minimal portions derived from non-human antibodies. Humanised antibodies include antibodies in which complementary determining regions (CDRs) of a human antibody (recipient antibody) are replaced by residues from a complementary determining region of interest, from a non-human species (donor antibody) such as mouse, rat or rabbit having the desired functionality. In some instances, Fv framework residues of the human antibody are replaced by corresponding non-human residues. Humanised antibodies may also comprise residues which are found neither in the recipient antibody nor in the imported complementarity determining region or framework sequences. In general, the humanised antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the complementarity determining regions correspond to those of a donor antibody of interest, such as a non- human antibody and all, or substantially all, of the framework regions correspond to those of a relevant human consensus sequence. Humanised antibodies optimally also include at least a portion of an antibody constant region, such as an Fc region, typically derived from a human antibody (see, for example, Jones et al., 1986. Nature 321 : 522- 525; Riechmann et al., 1988, Nature 332:323-329; Presta, 1992, Curr. Op. Struct. Biol. 2: 593-596, which are incorporated herein by reference).

[0443] Methods for humanising non-human antibodies are well known in the art. Generally, the humanised antibody has one or more amino acid residues introduced into it from a source which is non-human. These non-human amino acid residues, often referred to as imported residues, are typically taken from an imported variable domain. Humanisation can be essentially performed as described (see, for example, Jones et al., 1986; Reichmann et a / ., 1988; Verhoeyen etal., 1988, Science 239: 1534- 15361; US 4,816,567, which are incorporated herein by reference) by substituting human complementarity determining regions with corresponding rodent complementarity determining regions. Accordingly, such humanised antibodies are chimeric antibodies, wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanised antibodies may be typically human antibodies in which some complementarity determining region residues and possibly some framework residues are substituted by residues from analogous sites in rodent antibodies.

[0444] Human antibodies can also be identified using various techniques known in the art, including phage display libraries (see, for example, Hoogenboom & Winter, 1991, J. Mol. Biol. 227:381; Marks et al., 1991, J. Mol. Biol. 222: 581; Cole et al., 1985, In: Monoclonal antibodies and Cancer Therapy, Alan R. Liss, pp. 77; Boerner et a / ., 1991. J. Immunol. 147:86-95, Soderlind et al., 2000, Nat Biotechnol 18:852-6 and WO 98 / 32845, which are incorporated herein by reference).

[0445] It will be appreciated by persons skilled in the art that the polypeptides, e.g. antibodies, of the present invention may be of any suitable structural format. Antibodies may also be comprised of an Fc region, for which there are Fc-specific receptors. Engineering the Fc region of a therapeutic monoclonal antibody or Fc fusion protein allows the generation of molecules that are better suited to the pharmacology activity required of them (Strohl, 2009, Curr Opin Biotechnol 20(6): 685-91, the disclosures of which are incorporated herein by reference).

[0446] (a) Engineered Fc regions for increased half-life

[0447] One approach to improve the efficacy of a therapeutic antibody is to increase its serum persistence, thereby allowing higher circulating levels, less frequent administration and reduced doses.

[0448] The half-life of an IgG depends on its pH-dependent binding to the neonatal receptor FcRn. FcRn, which is expressed on the surface of endothelial cells, binds the IgG in a pH-dependent manner and protects it from degradation.

[0449] Some antibodies that selectively bind the FcRn at pH 6.0, but not pH 7.4, exhibit a higher half-life in a variety of animal models.

[0450] Several mutations located at the interface between the CH2 and CH3 domains, such as T250Q / M428L (Hinton et al., 2004, J Biol Chem. 279(8): 6213-6, the disclosures of which are incorporated herein by reference) and M252Y / S254T / T256E + H433K / N434F (Vaccaro et al., 2005, Nat Biotechnol. 23(10) : 1283-8, the disclosures of which are incorporated herein by reference), have been shown to increase the binding affinity to FcRn and the half-life of IgGl in vivo.

[0451] (b) Engineered Fc regions for altered effector function

[0452] The affinity of the Fc portion of an antibody to FcyRI, FcyRII and III can be compared with wild-type IgGl by flow cytometry, to determine by what concentration half maximal binding is achieved of FcyR expressing cells (Hezareh etal., 2001, J Virol., 75(24) : 12161-12168, incorporated herein by reference). This can alternatively be determined by FcyR enzyme linked immunosorbent assays (ELISA) (Shields et al., 2001, Mol. Basis Cell Dev. Biol., 276(9) : 6591-6604, incorporated herein by reference).

[0453] The four human IgG isotypes bind the activating Fey receptors (FcyRI, FcyRIIa, FcyRIIIa), the inhibitory FcyRIIb receptor, and the first component of complement (Clq) with different affinities, yielding very different effector functions (Bruhns et al., 2009, Blood. 113(16) : 3716-25, the disclosures of which are incorporated herein by reference). IgGl molecules have the highest affinity and capacity to induce effector functions, whereas IgG2, IgG3 and IgG4 are less effective (Bruhns, 2012, Blood. 119(24): 5640-9; Hogarth and Pietersz, 2012, Nature Reviews Drug Discovery 11, 311- 331; Stewart et al. 2014 Journal for ImmunoTherapy of Cancer 2:29; Wang et al. 2015 Front Immunol; 6: 368; Vidarsson et al. 2014, Front Immunol. 5: 520, incorporated herein by reference). In addition, certain mutations in the Fc region of IgGl dramatically reduce FcyR affinity and effector function while retaining neonatal FcR (FcRn) interaction (Ju and Jung, 2014, Curr Opin Biotechnol. 30: 128-39; Leabman et al. 2013, mAbs, 5:6, 896-903; Oganesyan et al. 2008 Acta Crystallogr D Biol Crystallogr. 64(Pt 6) : 700-704; Oganesyan et al. 2008 Mol Immunol. 45(7) : 1872-82; Sazinsky et al., 2008 Proc. Natl. Acad. Sci. U.S.A 105(51) 20167-20172, the disclosures of which are incorporated herein by reference).

[0454] The most widely used IgGl mutants are N297A alone or in combination with D265A, as well as mutations at positions L234 and L235, including the so-called "LALA" double mutant L234A / L235A. Another position described to further silence IgGl by mutation is P329 (see US 2012 / 0251531). Additional mutations in the Fc region are described in WO 2021 / 234402. Accordingly, binding molecules of the present invention may incorporate any of the Fc regions as described in WO 2021 / 234402, the contents of which are incorporated herein by reference.

[0455] In exemplary embodiments, the polypeptide is selected from the groups consisting of (any of which may be monospecific or bispecific) :

[0456] (a) bivalent antibodies, such as IgG-scFv antibodies (for example, wherein a first binding domain is an intact IgG and a second binding domain is an scFv attached to first binding domain at the N-terminus of a light chain and / or at the C-terminus of a light chain and / or at the N-terminus of a heavy chain and / or at the C-terminus of a heavy chain of the IgG, or vice versa) ;

[0457] (b) monovalent antibodies, such as a DuoBody® (Genmab AS, Copenhagen, Denmark) or 'knob-in-hole' bispecific antibody (for example, an scFv-KIH, scFv-KIHr, a BiTE-KIH or a BiTE-KIHr(see Xu et a!., 2015, mAbs 7(l) :231-242);

[0458] (c) scFvz-Fc antibodies (such as ADAPTIR™ bispecific antibodies from Emergent Biosolutions Inc);

[0459] (d) BiTE / scFvz antibodies;

[0460] (e) DVD-Ig antibodies;

[0461] (f) DART-based antibodies (for example, DART2-FC or DART);

[0462] (g) DNL-Fabs antibodies; and

[0463] (h) scFv-HSA-scFv antibodies.

[0464] For example, the antibody may be an IgG-scFv antibody. The IgG-scFv antibody may be in either VH-VL or VL-VH orientation. In one embodiment, the scFv may be stabilised by a S-S bridge between VH and VL.

[0465] A first binding domain and second binding domain may be fused directly to each other. Alternatively, a first binding domain and second binding domain may be joined via a linker, such as a polypeptide linker. For example, a polypeptide linker may be a short linker peptide between about 10 to about 25 amino acids. The linker is usually rich in glycine for flexibility, as well as serine or threonine for solubility, and can either connect the N-terminus of the VH with the C-terminus of the VL, or vice versa.

[0466] The terms "binding activity" and "binding affinity" are intended to refer to the tendency of a polypeptide molecule to bind or not to bind to a target. Binding affinity may be quantified by determining the dissociation constant (Kd) for a polypeptide and its target. A lower Kd is indicative of a higher affinity for a target. Similarly, the specificity of binding of a polypeptide to its target may be defined in terms of the comparative dissociation constants (Kd) of the polypeptide for its target as compared to the dissociation constant with respect to the polypeptide and another, non-target molecule.

[0467] The value of this dissociation constant can be determined directly by well-known methods, and can be computed even for complex mixtures by methods such as those, for example, set forth in Caceci et al., 1984. For example, the Kd may be established using a double-filter nitrocellulose filter binding assay such as that disclosed by Wong & Lohman, 1993. Other standard assays to evaluate the binding ability of ligands such as antibodies towards targets are known in the art, including for example, ELISAs, Western blots, RIAs, and flow cytometry analysis. The binding kinetics (e.g. binding affinity) of the polypeptide also can be assessed by standard assays known in the art, such as by Biacore™ system analysis.

[0468] A competitive binding assay can be conducted in which the binding of the polypeptide to the target is compared to the binding of the target by another, known ligand of that target, such as another polypeptide. The concentration at which 50% inhibition occurs is known as the Ki. Under ideal conditions, the Ki is equivalent to Kd. The Ki value will never be less than the Kd, so measurement of Ki can conveniently be substituted to provide an upper limit for Kd.

[0469] Alternative measures of binding affinity include EC50 or IC50. In this context EC50 indicates the concentration at which a polypeptide achieves 50% of its maximum binding to a fixed quantity of target. IC50 indicates the concentration at which a polypeptide inhibits 50% of the maximum binding of a fixed quantity of competitor to a fixed quantity of target. In both cases, a lower level of EC50 or IC50 indicates a higher affinity for a target. The EC50 and IC50 values of a ligand for its target can both be determined by well-known methods, for example ELISA. Suitable assays to assess the EC50 and IC50 of polypeptides are set out in the Examples.

[0470] As mentioned, antibodies may be produced by standard techniques, for example by immunisation with the appropriate (glyco)polypeptide or portion(s) thereof, or by using a phage display library.

[0471] If polyclonal antibodies are desired, a selected mammal (e.g. mouse, rabbit, goat, horse, etc) is immunised with an immunogenic polypeptide bearing a desired epitope(s), optionally haptenised to another polypeptide. Depending on the host species, various adjuvants may be used to increase immunological response. Such adjuvants include, but are not limited to, Freund's, mineral gels such as aluminium hydroxide, and surface-active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpet hemocyanin, and dinitrophenol. Serum from the immunised animal is collected and treated according to known procedures. If serum containing polyclonal antibodies to the desired epitope contains antibodies to other antigens, the polyclonal antibodies can be purified by immunoaffinity chromatography. Techniques for producing and processing polyclonal antisera are well known in the art.

[0472] Monoclonal antibodies directed against entire polypeptides or particular epitopes thereof (for example, in the case of the present application, against the ECD3 of US28) can also be readily produced by one skilled in the art. The general methodology for making monoclonal antibodies by hybridomas is well known. Immortal antibody-producing cell lines can be created by cell fusion, and also by other techniques such as direct transformation of B lymphocytes with oncogenic DNA, or transfection with Epstein-Barr virus. Panels of monoclonal antibodies produced against the polypeptides listed above can be screened for various properties; i.e. for isotype and epitope specificity and / or affinity, as well as strain agnostic binding properties. Monoclonal antibodies may be prepared using any of the well-known techniques which provides for the production of antibody molecules by continuous cell lines in culture.

[0473] In some embodiments, it may be preferred if the antibody is a monoclonal antibody. In some circumstance, particularly if the antibody is to be administered repeatedly to a human patient, it is preferred if the monoclonal antibody is a human monoclonal antibody or a humanised monoclonal antibody, which are suitable for administration to humans without engendering an immune response by the human against the administered immunoglobulin. Suitably prepared non-human antibodies can be "humanised" in known ways, for example by inserting the CDR regions of mouse antibodies into the framework of human antibodies. Humanised antibodies can be made using the techniques and approaches described in Verhoeyen et al., (1988) Science, 239, 1534-1536, and in Kettleborough et al., (1991) Protein Engineering, 14(7), 773- 783. In some instances, Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues. In general, the humanised antibody will contain variable domains in which all or most of the CDR regions correspond to those of a non-human immunoglobulin, and framework regions which are substantially or completely those of a human immunoglobulin consensus sequence.

[0474] Completely human antibodies may be produced using recombinant technologies. Typically, large libraries comprising billions of different antibodies are used. In contrast to the previous technologies employing chimerisation or humanisation of, e.g., murine antibodies, this technology does not rely on immunisation of animals to generate the specific antibody. Instead the recombinant libraries comprise a huge number of pre-made antibody variants wherein it is likely that the library will have at least one antibody specific for any antigen. Thus, using such libraries, an existing antibody having the desired binding characteristics can be identified. In order to find the good binder in a library in an efficient manner, various systems where phenotype, i.e. the antibody or fragment thereof, is linked to its genotype, i.e. the encoding gene(s), have been devised. The most commonly used such system is the so-called phage display system where antibody fragments are expressed, displayed, as fusions with phage coat proteins on the surface of filamentous phage particles, while simultaneously carrying the genetic information encoding the displayed molecule (McCafferty et al., 1990, Nature 348: 552-554). Phage displaying antibody fragments specific for a particular antigen may be selected through binding to the antigen in question. Isolated phage may then be amplified and the gene encoding the selected antibody variable domains may optionally be transferred to other antibody formats, such as e.g. full-length immunoglobulin, and expressed in high amounts using appropriate vectors and host cells well known in the art. Alternatively, the "human" antibodies can be made by immunising transgenic mice which contain, in essence, human immunoglobulin genes (Vaughan et al., (1998) Nature Biotechnol. 16, 535-539).

[0475] It is appreciated that when the antibody is for administration to a non-human individual, the antibody may have been specifically designed / produced for the intended recipient species.

[0476] The format of displayed antibody specificities on phage particles may differ. The most commonly used formats are Fab (Griffiths et al., 1994. EMBO J. 13: 3245-3260) and scFv (Hoogenboom et al., 1992, J Mol Biol. 227: 381-388) both comprising the variable antigen binding domains of antibodies. The single chain format is composed of a variable heavy domain (VH) linked to a variable light domain (VL) via a flexible linker (US 4,946,778). Before use as a therapeutic agent, the antibody may be transferred to a soluble format e.g. Fab or scFv and analysed as such. In later steps the antibody fragment identified to have desirable characteristics may be transferred into yet other formats such as full-length antibodies.

[0477] WO 98 / 32845 and Soderlind et al., (2000) Nature BioTechnol. 18: 852-856 describe technology for the generation of variability in antibody libraries. Antibody fragments derived from this library all have the same framework regions and only differ in their CDRs. Since the framework regions are of germline sequence the immunogenicity of antibodies derived from the library, or similar libraries produced using the same technology, are expected to be particularly low (Soderlind et al., 2000, supra). This property is of great value for therapeutic antibodies, reducing the risk that the patient forms antibodies to the administered antibody, thereby reducing risks for allergic reactions, the occurrence of blocking antibodies, and allowing a long plasma half-life of the antibody. Thus, when developing therapeutic antibodies to be used in humans, modern recombinant library technology (Soderlind et al., 2001, Comb. Chem. & High Throughput Screen. 4: 409-416) is now used in preference to the earlier hybridoma technology.

[0478] By antibodies we also include heavy-chain antibodies structurally derived from camelidae antibodies, such as Nanobodies® (Ablynx). These are antibody-derived therapeutic proteins that contain the structural and functional properties of naturally- occurring heavy-chain antibodies. The Nanobody® technology was developed following the discovery that camelidae (camels and llamas) possess fully functional antibodies that lack light chains. These heavy-chain antibodies contain a single variable domain (VHH) and two constant domains (CH2 and CH3) . The cloned and isolated VHH domain is a perfectly stable polypeptide harbouring the full antigen-binding capacity of the original heavy-chain antibody. These VHH domains with their unique structural and functional properties form the basis of Nanobodies®. They combine the advantages of conventional antibodies (high target specificity, high target affinity and low inherent toxicity) with important features of small molecule drugs (the ability to inhibit enzymes and access receptor clefts). Furthermore, they are stable, have the potential to be administered by means other than injection, are easier to manufacture, and can be humanised. (See, for example US 5,840,526; US 5,874,541; US 6,005,079, US 6.765,087; EP 1 589 107; WO 97 / 34103; WO97 / 49805; US 5,800,988; US 5,874, 541 and US 6,015,695).

[0479] It is preferred that the antibody, or other binding molecule, that selectively binds to ECD3 of US28 does not bind a related polypeptide, such as CCR5, or that the antibody binds US28 with a greater affinity than for the related polypeptide, such as CCR5. Preferably, the antibody binds the US28 with at least 5, or at least 10 or at least 50 times greater affinity than for the related polypeptide. More preferably, the antibody molecule binds the US28 with at least 100, or at least 1,000, or at least 10,000 times greater affinity than for the related polypeptide. Such binding may be determined by methods well known in the art, such as one of the Biacore® systems.

[0480] Bispecific Antibodies and other Bispecific Binding Molecules

[0481] In one embodiment of particular interest, the binding molecule of the first aspect of the present invention is, or comprises, a bispecific binding molecule.

[0482] For example, a bispecific binding molecule in accordance with the first aspect of the present invention may comprise a first domain capable of recruiting the activity of an effector cell by specifically binding to an effector antigen located on the effector cell; and a second domain capable of specifically binding to ECD3 of the HCMV-encoded US28 protein, as a target antigen, wherein said target antigen may be located on a target cell other than the effector cell. The second domain is, or comprises, a binding molecule according to the first aspect of the present invention.

[0483] All or part of the first and second domains may optionally be connected by one or more linker sequences. The first domain may comprise multiple functional domains and each of those functional domains may be connected to a neighbouring domain either directly or via a linker sequence and / or may be formed from more than one separate polypeptide sequence. The second domain may comprise multiple functional domains and each of those functional domains may be connected to a neighbouring domain either directly or via a linker sequence and / or may be formed from more than one separate polypeptide sequence.

[0484] In the instances in which one or more linker sequence is present in a bispecific binding molecule of the present invention, then each linker sequence may independently be selected from any suitable linker sequence. For example, each linker sequence may be at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, or more amino acid residues in length. Each linker sequence may comprise any naturally occurring amino acid. In some embodiments, each linker sequence may comprise or consist of the amino acids glycine and serine. In another embodiment, the or each linker sequence may comprise or consist of sets of glycine and serine repeats such as (Gly4Ser)n, where n is a positive integer equal or greater than 1, such as 2, 3, 4, 5, 6 or more. In one embodiment, the linker is (Gly4Ser)3. Variation in the linker length may retain or enhance activity, giving rise to superior efficacy in activity studies.

[0485] In one embodiment, the bispecific binding molecule is a bispecific antibody (bsAb) format, such as those described in Suurs et al., 2019, Pharmacology & Therapeutics, 201 : 103-119 (the content of which are incorporated herein by reference), and in particular one or more of the formats illustrated in Figure 1C of Suurs et al., 2019 (supra), such as a TriMab, an IgG-like BsAb, a CrossMab, a 2: 1 CrossMab, a 2:2 CrossMab, a DuoBody, a DVD-Ig BsAb, an scFv-IgG, and IgG-IgG, a Fab-scFv-Fc, a TF, an ADAPTIR, a BiTE, a BiTE-Fc, a DART, a DART-Fc, a Tetravalent DART, a TandAb, an ImmTAC, a TriKE, and scFv-scFv-scFv, or a tris-specific nanobody format.

[0486] In one embodiment, the first and / or second domain may comprise at least one variable light chain and / or at least one variable heavy chain. In one embodiment, the first and / or second domain may comprise at least one variable light chain and at least one variable heavy chain to form an scFv. In one embodiment, the first and / or second domain comprise a noncovalent dimer of scFv connected by a linker (e.g. a small peptide linker) to form a diabody. In one embodiment, the first and / or second domain may comprise multiple diabodies to form a tandem diabody (TandAb).

[0487] In one embodiment, the first and / or second domain may further comprise at least one constant light chain and / or at least one constant heavy chain. For example, the first and / or second domain may comprise an scFv that further comprises a constant light chain and a constant heavy chain to form a Fab. In one embodiment, the first and / or second domain comprises multiple Fab (for example, two Fab).

[0488] In one embodiment, the first and / or second domain may further comprise at least one Fc. For example, the first and / or second domain may comprise an scFv that further comprises an Fc to form an scFv-IgG. Additionally, or alternatively, the first and / or second domain may comprise two Fab domains that further comprises an Fc to form an IgG.

[0489] In one embodiment, the first domain may be an scFv connected to a second domain that is an scFv, optionally via a linker, to form a BiTE, optionally wherein the BiTE further comprises an Fc to form a BiTE-Fc.

[0490] In one embodiment, the VH or VL of the first domain may be swapped with the VH or VL of the second domain, respectively, to form a DART comprising a first and second domain, optionally wherein the DART further comprises an Fc to form a DART- Fc. Additionally, or alternatively, multiple DARTs may be connected to a single Fc to form a tetravalent DART.

[0491] In one embodiment, the first and / or second domain is in the form of a T cell receptor, which may be combined with a first and / or second domain that is in the form of an scFv to form an ImmTAC. For example, the first domain may be an scFv and the second domain may be a T cell receptor, preferably wherein the scFv is specific for an epitope of CD3 (i.e. a CD3-binding domain), thereby resulting in an ImmTAC.

[0492] In one embodiment, the effector cell of the first domain is an immune effector cell, for example an immune effector cell selected from the group consisting of a T cell (for example, a CD4+T cell and / or a CD8+T cell), NK T cell, NK cell, macrophage, or any recombinant cell thereof (for example, a CAR-expressing cells, such as a CAR-T cell, a CAR-NK cell or a CAR-M cell). In such embodiments, the bispecific binding molecule may be referred to as a 'bispecific immune cell engager antibody', wherein the "immune cell" may be substituted for the effector cell type. For example, if the effector cell of the first domain is a T cell, such embodiments may be referred to as a 'bispecific T cell engager antibody' (BiTE, which may, but does not necessarily have the format of the BiTE or BiTE-Fc molecule shown in Fig 1C of Suurs et al, 2019, supra), and if the effector cell of the first domain is an NK cell, such embodiments may be referred to as a 'bispecific NK cell engager antibody' and so on. In one embodiment, the first domain of the bispecific binding molecule is specific for an epitope of CD3, which may be referred to as a CD3-binding domain.

[0493] In one embodiment, the CD3-binding domain may comprise the sequence of OKT3 heavy chain variable region, for example as defined by the sequence of SEQ ID NO: 48 of the present application, or a functional variant or fragment thereof which substantially retains the CD3-binding specificity and / or affinity of the sequence of SEQ ID NO: 48. The functional variant or fragment thereof may optionally comprises one, two or three CDRs corresponding to any one, two or all three of the CDR sequences of the OKT3 heavy chain variable region as defined by SEQ ID NO: 48. The CDR sequences of OKT3 heavy chain variable region are as defined by SEQ ID NOs: 49, 50 and 51, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody OKT3.

[0494] In another embodiment, the CD3-binding domain may comprise the sequence of OKT3 light chain variable region, for example as defined by the sequence of SEQ ID NO: 52 of the present application, or a functional variant or fragment thereof which substantially retains the CD3-binding specificity and / or affinity of the sequence of SEQ ID NO: 52. The functional variant or fragment thereof may optionally comprises one, two or three CDRs corresponding to any one, two or all three of the CDR sequences of the OKT3 light chain variable region as defined by SEQ ID NO: 52. The CDR sequences of OKT3 heavy chain variable region are as defined by SEQ ID NOs: 53, 54 and 55, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody OKT3.

[0495] In another embodiment, the CD3-binding domain may comprise both of the sequences of OKT3 heavy chain variable region and the OKT3 light chain variable region, for example as defined by the sequences of SEQ ID NOs: 48 and 52 of the present application, respectively, or a functional variant or fragment thereof of either or both, such as a functional variant or fragment as described above.

[0496] In a preferred embodiment, the CD3-binding domain may comprise both of the sequences of OKT3 heavy chain variable region and the OKT3 light chain variable region, for example as defined by the sequences of SEQ ID NOs: 48 and 52 of the present application, optionally joined by a linker, such as a linker having the sequence GGGGSGGGGSGGGGS (SEQ ID NO: 56). This combination of an OKT3 heavy chain variable region and an OKT3 light chain variable region, optionally joined by a linker, may form an scFv of OKT3.

[0497] Accordingly, the CD3-binding domain may be an scFv of OKT3 that comprises the sequences of SEQ ID Nos: 48, 56 and 52, joined together in that order, wherein the OKT3 heavy chain variable region of SEQ ID NO: 48 is joined at its C-terminus to the N-terminus of the linker sequence of SEQ ID NO: 56, which in turn is joined at its C-terminus to the N-terminus of the OKT3 light chain variable region of SEQ ID NO: 52. Alternatively, the order of the light and heavy chain regions may be swapped around, so that the CD3-binding domain comprises the sequences of SEQ ID Nos: 52, 56 and 48, joined together in that order. It will be appreciated that other linker sequences may be used in place of the exemplary sequence of SEQ ID NO: 56; other linker sequences may optionally comprise multiple repeats of the sequence GGGGS (SEQ ID NO: 57), and thus may have the sequence [GGGGS]n, wherein n is an integer of 2, 3, 4, 5, 6, 7, 8, 9 10 or more; or may contain any other suitable linker sequences.

[0498] Optionally, the CD3-binding domain comprises at least one (preferably two) scFvs of OKT3. For example, the bispecific binding molecule may comprise a first domain comprising two scFvs of OKT3, optionally wherein the two scFv are linked directly to each other or linked indirectly via the second domain.

[0499] In one embodiment, the second domain of the bispecific binding molecule, which comprises a binding molecule according to the first aspect of the present invention and has binding specificity to ECD3 of the US28 protein of HCMV, has binding specificity for a target cell that is a US28-expressing cell and / or a HCMV infected cell (which may, for example, be latently infected or lytically infected).

[0500] In one embodiment, the second domain of the bispecific binding molecule comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody:

[0501] (a) 1D3 as defined by SEQ ID NOs: 8, 9, 10, 14, 15 and 16, respectively;

[0502] (b) 1C10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively;

[0503] (c) 1A10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively;

[0504] (d) 1G9 as defined by SEQ ID NOs: 76, 77, 78, 82, 83 and 84, respectively; and / or

[0505] (e) 1E8 as defined by SEQ ID NOs: 76, 95, 96, 82, 99 and 100, respectively; and / or a functional variant of any one or more of said CDR sequences of antibody 1D3, 1C10, 1A10, 1G9 and / or 1E8.

[0506] Optionally, the US28-binding domain according to this embodiment may comprise:

[0507] (a) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable heavy chain (VH) of antibody:

[0508] I. 1D3, as defined by SEQ ID NOs: 8, 9, and 10, respectively; ii. 1C10, as defined by SEQ ID NOs: 112, 113, and 114, respectively (and / or a functional variant of any one, two or three of said CDR sequences); ill. 1A10, as defined by SEQ ID NOs: 112, 113, and 114, respectively (and / or a functional variant of any one, two or three of said CDR sequences); iv. 1G9, as defined by SEQ ID NOs: 76, 77, and 78, respectively (and / or a functional variant of any one, two or three of said CDR sequences); and / or v. 1E8, as defined by SEQ ID NOs: 76, 95, and 96, respectively (and / or a functional variant of any one, two or three of said CDR sequences); and / or

[0509] (b) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable light chain (VL) of antibody: i. 1D3, as defined by SEQ ID NOs: 14, 15, and 16, respectively (and / or a functional variant of any one, two or three of said CDR sequences);

[0510] II. 1C10, as defined by SEQ ID NOs: 117, 83, and 118, respectively (and / or a functional variant of any one, two or three of said CDR sequences); ill. 1A10, as defined by SEQ ID NOs: 117, 83, and 118, respectively (and / or a functional variant of any one, two or three of said CDR sequences); iv. 1G9, as defined by SEQ ID NOs: 82, 83, and 84, respectively (and / or a functional variant of any one, two or three of said CDR sequences); and / or v. 1E8, as defined by SEQ ID NOs: 82, 99, and 100, respectively (and / or a functional variant of any one, two or three of said CDR sequences).

[0511] Preferably, the US28-binding domain according to this embodiment comprises at least 4 different CDR sequences, for example at least 5 or 6 different CDR sequences.

[0512] Additionally, or alternatively, in a further option the second domain of the bispecific binding molecule according to this embodiment may comprise: (a) at least one variable heavy chain (VH) polypeptide that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID NOs: 8, 9, and 10, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 12; and / or (b) at least one variable light chain (VL) polypeptide that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 14, 15, and 16, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 18.

[0513] Additionally, or alternatively, in a further option the second domain of the bispecific binding molecule according to this embodiment may comprise: (a) at least one variable heavy chain (VH) polypeptide that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID NOs: 76, 77, and 78, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 68; and / or (b) at least one variable light chain (VL) polypeptide that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 82, 83, and 84, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 72.

[0514] Additionally, or alternatively, in a further option the second domain of the bispecific binding molecule according to this embodiment may comprise: (a) at least one variable heavy chain (VH) polypeptide that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID NOs: 76, 95, and 96, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 88; and / or (b) at least one variable light chain (VL) polypeptide that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 82, 99, and 100, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 92.

[0515] Additionally, or alternatively, in a further option the second domain of the bispecific binding molecule according to this embodiment may comprise: (a) at least one variable heavy chain (VH) polypeptide that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID NOs: 112, 113, and 114, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 104; and / or (b) at least one variable light chain (VL) polypeptide that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 117, 83, and 118, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 108.

[0516] Additionally, or alternatively, in a further option the second domain of the bispecific binding molecule according to this embodiment may comprise: (a) at least one variable heavy chain (VH) polypeptide that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID NOs: 112, 113, and 114, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 122; and / or (b) at least one variable light chain (VL) polypeptide that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 117, 83, and 118, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 126.

[0517] Additionally, or alternatively, in a further option the second domain of the bispecific binding molecule according to this embodiment may comprise: (a) at least one variable heavy chain (VH) polypeptide that comprises CDR 1, 2, and 3 sequences having the sequences of SEQ ID NOs: 138, 139, and 140, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 130; and / or (b) at least one variable light chain (VL) polypeptide that comprises CDR 1, 2 and 3 sequences having the sequences of SEQ ID NOs: 144, 145, and 146, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 134.

[0518] In one embodiment, the first domain of the bispecific binding molecule is an scFv comprising all six CDR sequences of OKT3 (preferably wherein the first domain is an scFv of OKT3), and the second domain of the bispecific binding molecule is an scFv comprising all six CDR sequences of 1D3, 1C10, 1A10, 1G9 and / or 1E8 (preferably wherein the second domain is an scFv of 1D3, 1C10, 1A10, 1G9 and / or 1E8), optionally wherein the first and second domain are connected via a linker.

[0519] In one embodiment, the first domain of the bispecific binding molecule comprises two scFv each comprising all six CDR sequences of OKT3 (preferably wherein the first domain comprises two scFv of OKT3); and the second domain of the bispecific binding molecule comprises two Fab each comprising one, two, three, four, five or six CDRs corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody:

[0520] (a) 1D3 as defined by SEQ ID NOs: 8, 9, 10, 14, 15 and 16 (preferably wherein the first domain comprises two Fab of 1D3);

[0521] (b) 1C10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118 (preferably wherein the first domain comprises two Fab of 1C10);

[0522] (c) 1A10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118 (preferably wherein the first domain comprises two Fab of 1A10);

[0523] (d) 1G9 as defined by SEQ ID NOs: 76, 77, 78, 82, 83 and 84 (preferably wherein the first domain comprises two Fab of 1G9); and / or

[0524] (e) 1E8 as defined by SEQ ID NOs: 76, 95, 96, 82, 99 and 100 (preferably wherein the first domain comprises two Fab of 1E8); and further comprises an Fc, optionally wherein the Fc corresponds to the Fc of human IgG. In some embodiments, the two Fab are identical. In some embodiments, the two Fab correspond to different antibodies selected from the group consisting of 1D3, 1C10, 1A10, 1G9 and 1E8.

[0525] In one particular embodiment, the bispecific binding molecule is a BiTE having a CD3-binding OKT3 scFv domain binding region, and comprises a US28 ECD3-binding region having the sequence of the 1D3 antibody, and more specifically preferably comprises, consists essentially of, or consists of:

[0526] (I) a first polypeptide sequence comprising, consisting essentially of, or consisting of a heavy chain sequence that includes the VH region of 1D3, optionally one or more CH sequences (such as a CHI, CH2 and / or CH3 sequence), an optional linker sequence, and the sequence of an OKT3 scFv, for example a heavy chain sequence having the sequence of SEQ ID NO: 58; and

[0527] (ii) a second polypeptide sequence comprising, consisting essentially of, or consisting of a light chain sequence that includes the VL region of 1D3, and optionally a CL sequence, for example a light chain sequence having the sequence of SEQ ID NO: 59.

[0528] In another embodiment, the sequence of the 1D3 antibody may be replaced in the BiTE by the corresponding sequences of an antibody selected from the group consisting of 1C10, 1A10, 1G9 and 1E8.

[0529] CARs / CAR T cells and other cells recombinantly expressing CARs

[0530] Immunotherapy-based treatments can be based on Chimeric Antigen Receptors (CARs). CARs are recombinant receptors for antigen, which, in a single molecule, redirect the specificity and function of T lymphocytes and other immune cells (Sadelain et al 2013 Cancer Discov 3(4) : 388).

[0531] The general premise for their use in cancer immunotherapy is to rapidly generate tumour-targeted T cells, bypassing the barriers and incremental kinetics of active immunisation. Once expressed in T cells, CAR-modified T cells acquire supra- physiological properties and act as "living drugs" that may exert both immediate and long-term effects. The engineering of CARs into T cells requires that T cells be cultured to allow for transduction and expansion. The transduction may utilise a variety of methods, but stable gene transfer is required to enable sustained CAR expression in clonally expanding and persisting T cells.

[0532] In principle, any cell surface molecule can be targeted through a CAR, thus overriding tolerance to self-antigens and the antigen recognition gaps in the physiological T cell repertoire that limit the scope of T cell reactivity.

[0533] Various T cell subsets, as well as T cell progenitors and other immune cells such as natural killer cells, can be targeted with a CAR. Adoptive immunotherapy using CAR engineered cells such as T cells is a promising approach in cancer treatment (Han et al 2013 J Hematol Oncol 6: 47). Significant progresses made in the past decades have contributed to the development of more efficient antitumour immunotherapy. For example, incorporation of a single chain variable fragment (scFv) of a tumour antigen specific antibody and signalling domains of T cell receptor renders CARs having the specificity of an antibody and the cytotoxicity of cytotoxic T lymphocytes. CARs endow T cells antigen specific recognition, activation and proliferation in an MHC independent manner. In addition, CAR bypasses many mechanisms through which cancer cells escape immunorecognition. These mechanisms include down-regulation of the MHC, reduced expression of costimulatory molecules, induction of suppressive cytokines and recruitment of regulatory T cells. Besides these beneficial effects, the technical feasibility of CARs make them even more attractive in the development of adoptive immunotherapy. The observations from preclinical and clinical studies have revealed a very encouraging therapeutic efficacy of CAR-mediated immunotherapy in a variety of cancers including lymphoma, chronic lymphocytic leukaemia, melanoma and neuroblastoma.

[0534] In one embodiment, the binding molecule of the first aspect of the present invention can be a CAR, for example a CAR comprising:

[0535] (I) an extracellular domain, wherein the extracellular domain comprises or consists of a binding molecule (such as an antibody) as defined above, or a functional fragment of said binding molecule;

[0536] (ii) a transmembrane domain; and

[0537] (iii) an intracellular domain; wherein the extracellular domain of the CAR has binding specificity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of human cytomegalovirus (HCMV), and wherein ECD3 of the US28 protein comprises an amino acid sequence presented in the US28 protein at positions corresponding to positions 167 to 183 of the US28 protein encoded by human cytomegalovirus (HCMV) as set forth in SEQ ID NO: 5.

[0538] Optionally:

[0539] (a) the extracellular domain of the CAR has binding specificity to an epitope present entirely within extracellular domain 3 (ECD3) of the US28 protein of HCMV;

[0540] (b) the extracellular domain of the CAR has binding specificity to a linear epitope within ECD3 of the US28 protein;

[0541] (c) the extracellular domain of the CAR has binding specificity to an epitope within ECD3 of a US28 protein of HCMV that is HCMV strain agnostic, for example, binding specificity to an epitope within ECD3 of a US28 protein of HCMV that is agnostic to two or more (such as all) of HCMV strains selected from the group consisting of DB, Towne, AD169, BL, DAVIS, JP, Merlin, PH, TB40 / E, Toledo, TR, VHL / E and VR1814 (FIX); and / or

[0542] (d) the extracellular domain of the CAR has specificity to an epitope within ECD3 of the US28 protein of HCMV, irrespective of whether the ECD3 of the US28 protein comprises the sequence of:

[0543] - TKKDNQCMTDYDYLEVS (SEQ ID NO: 7) as found in ECD3 of US28 as encoded by a majority of HCMV strains, or

[0544] - TKKNNQCMTDYDYLEVS (SEQ ID NO: 6) as found in ECD3 of US28 as encoded by a minority of HCMV strains.

[0545] In one preferred embodiment, the extracellular domain of the CAR may comprise one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody:

[0546] (a) 1D3 as defined by SEQ ID NOs: 8, 9, 10, 14, 15 and 16, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1D3;

[0547] (b) 1C10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1C10;

[0548] (c) 1A10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1A10;

[0549] (d) 1G9 as defined by SEQ ID NOs: 76, 77, 78, 82, 83 and 84, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1G9; and / or

[0550] (e) 1E8 as defined by SEQ ID NOs: 76, 95, 96, 82, 99 and 100, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1E8.

[0551] Additionally, or alternatively, the extracellular domain of the CAR may comprise:

[0552] (a) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable heavy chain (VH) of antibody: i. 1D3, as defined by SEQ ID NOs: 8, 9, and 10, respectively (or a functional variant of any one or more of said CDR sequences);

[0553] II. 1C10, as defined by SEQ ID NOs: 112, 113, and 114, respectively (or a functional variant of any one or more of said CDR sequences); Hi. 1A10, as defined by SEQ ID NOs: 112, 113, and 114, respectively (or a functional variant of any one or more of said CDR sequences); iv. 1G9, as defined by SEQ ID NOs: 76, 77, and 78, respectively (or a functional variant of any one or more of said CDR sequences); and / or v. 1E8, as defined by SEQ ID NOs: 76, 95, and 96, respectively (or a functional variant of any one or more of said CDR sequences); and / or

[0554] (b) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable light chain (VL) of antibody:

[0555] I. 1D3, as defined by SEQ ID NOs: 14, 15, and 16, respectively (or a functional variant of any one or more of said CDR sequences); ii. 1C10, as defined by SEQ ID NOs: 117, 83, and 118, respectively (or a functional variant of any one or more of said CDR sequences);

[0556] Hi. 1A10, as defined by SEQ ID NOs: 117, 83, and 118, respectively (or a functional variant of any one or more of said CDR sequences); iv. 1G9, as defined by SEQ ID NOs: 82, 83, and 84, respectively (or a functional variant of any one or more of said CDR sequences); v. 1E8, as defined by SEQ ID NOs: 82, 99, and 100, respectively (or a functional variant of any one or more of said CDR sequences).

[0557] Additionally, or alternatively, the extracellular domain of the CAR may comprise:

[0558] (a) at least one variable heavy chain (VH) polypeptide sequence that comprises CDR 1, 2, and 3 sequences having the sequences of: i. SEQ ID NOs: 8, 9, and 10, respectively (or a functional variant of any one or more of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 12 (or a functional variant thereof);

[0559] II. SEQ ID NOs: 112, 113, and 114, respectively (or a functional variant of any one or more of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 104 (or a functional variant thereof);

[0560] Hi. SEQ ID NOs: 112, 113, and 114, respectively (or a functional variant of any one or more of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 122 (or a functional variant thereof); iv. SEQ ID NOs: 76, 77, and 78, respectively (or a functional variant of any one or more of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 68 (or a functional variant thereof); and / or v. SEQ ID NOs: 76, 95, and 96, respectively (or a functional variant of any one or more of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 88 (or a functional variant thereof); and / or

[0561] (b) at least one variable light chain (VL) polypeptide sequence that comprises CDR 1, 2 and 3 sequences having the sequences of: i. SEQ ID NOs: 14, 15, and 16, respectively (or a functional variant of any one or more of said CDR sequences), and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 18 (or a functional variant thereof);

[0562] II. SEQ ID NOs: 117, 83, and 118, respectively (or a functional variant of any one or more of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 108 (or a functional variant thereof); ill. SEQ ID NOs: 117, 83, and 118, respectively (or a functional variant of any one or more of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 126 (or a functional variant thereof); iv. SEQ ID NOs: 82, 83, and 84, respectively (or a functional variant of any one or more of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 72 (or a functional variant thereof); and / or v. SEQ ID NOs: 82, 99, and 100, respectively (or a functional variant of any one or more of said CDR sequences), and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 92 (or a functional variant thereof). A functional variant of a reference sequence selected from SEQ ID NO: 12, 18, 68, 72, 88, 92, 104, 108, 122 or 126 may optionally have at least 70% sequence identity to the defined reference, for example at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%. 98% or 99% sequence identity, and more generally has 95-99% sequence identity to the defined amino acid sequence (e.g. 96%, 97%, 98% or 99% sequence identity). The variation may be solely within the non-CDR regions of the sequences of the functional variant of the variable heavy or light chain, solely within the CDR regions of the sequences, or within both the non-CDR and CDR regions of the sequences. It will be appreciated that, if variation is included in the CDR regions of the functional variant, then the variation may be within one CDR, two CDRs, or three CDRs, respectively of the heavy chain variable region, or the light chain variable region. Typically, the variation is outside of the CDR regions.

[0563] In certain embodiments, the extracellular domain of the CAR is, or comprises, consists essentially of, or consists of, the sequence of, an antibody, optionally a humanised antibody, for example a single-chain variable fragment (scFv) or a functional variant thereof, wherein said antibody and the functional variant thereof is a binding molecule according to the first aspect of the present invention.

[0564] An scFv can comprise a linker of at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, or more amino acid residues between its VL and VH regions. The linker may comprise any naturally occurring amino acid. In some embodiments, the linker sequence comprises amino acids glycine and serine. In another embodiment, the linker sequence comprises sets of glycine and serine repeats such as (Gly4Ser)n, where n is a positive integer equal or greater than 1, such as 2, 3, 4, 5, 6 or more. In one embodiment, the linker is (Gly4Ser)3. Variation in the linker length may retain or enhance activity, giving rise to superior efficacy in activity studies.

[0565] The light chain variable region and heavy chain variable region of a scFv can be, for example, in any of the following orientations: light chain variable region-linker- heavy chain variable region or heavy chain variable region-linker-light chain variable region.

[0566] A CAR as described herein comprises a transmembrane domain. By a transmembrane domain we include the meaning of any moiety that is capable of being embedded in a lipid membrane. By being embedded in a lipid membrane we include the meaning of the transmembrane domain favourably interacting with the hydrophobic portions of the lipids that make up the lipid membrane. Insertion into lipid membranes may be assayed using any suitable method known in the art, including fluorescence labelling with fluorescence microscopy. Hence, it will be appreciated that the transmembrane domain is one that locates the CAR molecule within the lipid membrane.

[0567] Optionally, the transmembrane domain comprises the transmembrane domain of a protein (e.g. a transmembrane protein), for example the transmembrane domain of a transmembrane receptor protein. In an embodiment, the transmembrane domain is one that is associated with one of the other domains of the CAR. In an embodiment, the transmembrane domain comprises the transmembrane portion of an intracellular signalling protein that constitutes at least part of the intracellular signalling domain. In some instances, the transmembrane domain can be selected or modified by amino acid substitution to avoid binding of such domains to the transmembrane domains of the same or different surface membrane proteins, e.g. to minimise interactions with other members of the receptor complex. In some instances, the transmembrane domain is capable of homodimerisation with another CAR on the cell surface.

[0568] The transmembrane domain may be derived either from a natural or from a recombinant source. The domain may be derived from any membrane-bound or transmembrane protein. In one embodiment, the transmembrane domain is capable of signalling to the intracellular domain(s) whenever the CAR has bound to a target. A suitable transmembrane domain for use in the invention may include the transmembrane region(s) of the alpha, beta or zeta chain of the T cell receptor, CD28, CD3 epsilon, CD8, CD45 and CD4.

[0569] The transmembrane domain can, for example, include one or more additional amino acids adjacent to the transmembrane region, such as one or more amino acids associated with the extracellular region of the protein from which the transmembrane domain was derived (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 up to 15 amino acids of the extracellular region) and / or one or more additional amino acids associated with the intracellular region of the protein from which the transmembrane protein is derived (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 up to 15 amino acids of the intracellular region).

[0570] In certain embodiments, the extracellular domain of the CAR is connected to the transmembrane domain by a hinge region. The hinge region may comprise one or more immunoglobulin domains. Particular examples include the Fc region of IgGl and the immunoglobulin-like extracellular regions of CD4 and CD8. It will be appreciated that when the antibody that selectively binds US28 is a scFv molecule, the hinge region can be used to extend the reach of the scFv to allows its attachment to the transmembrane domain without affecting its binding to US28. The hinge may be from a human protein such as human immunoglobulin. The hinge region may, for example, be a CD8o hinge region (for example, as described in An et al, 2016, Oncotarget, 7: 10638-10649, the contents of which are incorporated herein by reference). Typically, the transmembrane domain comprises predominantly hydrophobic amino acid residues such as leucine and valine.

[0571] In an embodiment, a short oligo- or polypeptide linker, such as between 2 and 10 amino acids in length, may form the linkage between the transmembrane domain and the intracellular signalling domain of the CAR. The linker may, for example, comprise glycine and / or serine residues. An example of a suitable linker is a glycineserine doublet.

[0572] By intracellular signalling domain we include the meaning of a domain that is capable of activating at least one of the normal functions of the cell in which the CAR is introduced, such as at least one of the normal effector functions of an immune cell (e.g. T cell). An effector function refers to a specialised function of a cell. The effector function of a T cell, for example, may be cytolytic function or helper activity including the secretion of cytokines. Thus, the intracellular signalling domain may be a portion of a protein which transduces the effector function signal and directs the cell (e.g. T cell) to perform a specialised function.

[0573] Generally, the whole intracellular signalling domain can be used; however, it is appreciated that it is not necessary to use the entire domain, provided that whatever part of the signalling domain that is used is still capable of transducing the effector function signal. It will also be appreciated that variants of such intracellular signalling domains with substantially the same or greater functional capability may also be used. By this we include the meaning that the variants should have substantially the same or greater transduction of the effector functional signal. Typically, substantially the same or greater signal transduction includes at least 80%, 85%, 90%, 95%, 100%, 105%, 110%, 115%, or 120%, or more of the signal transduction of the unmodified intracellular signalling domain, wherein signal transduction of the unmodified intracellular signalling domain corresponds to 100%.

[0574] Methods for assessing transduction of effector function signal are well known to those skilled in the art and include, for example, assessing the amounts and / or activity of molecules (e.g. proteins such as cytokines) that are indicative of the transduced signal. Thus, when the signal is the cytolytic function of a T-cell, the methods may involve measurement of one or more cytokines secreted by the T-cell, which cytokines are known to have a cytolytic activity (e.g. IFN gamma) and following measurement of the target cell lysis.

[0575] Examples of intracellular signalling domains for use in the CAR of the invention include the cytoplasmic sequences of the T cell receptor (TCR) and co-receptors that act in concert to initiate signal transduction following antigen receptor engagement, as well as any derivative or variant of these sequences and any recombinant sequence that has the same functional capability. It is known that signals generated through the TCR alone are generally insufficient for full activation of a T cell and that a secondary and / or costimulatory signal may also be required. Thus, T cell activation can be said to be mediated by two distinct classes of intracellular signalling sequences: those that initiate antigendependent primary activation through the TCR (primary intracellular signalling domains) and those that act in an antigen-independent manner to provide a secondary or costimulatory signal (secondary intracellular signalling domain, such as a costimulatory domain). Costimulatory domains promote activation of effector functions and may also promote persistence of the effector function and / or survival of the cell.

[0576] A primary intracellular signalling domain regulates primary activation of the TCR complex either in a stimulatory way, or in an inhibitory way. Primary intracellular signalling domains that act in a stimulatory manner may contain signalling motifs which are known as immunoreceptor tyrosine-based activation motifs or ITAMs (e.g. 2, 3, 4, 5 or more ITAMs). Thus, the intracellular signalling domain may comprise one or more ITAMs. Examples of ITAM containing primary intracellular signalling domains that are of particular use in the invention include those of CD3 zeta, Fc receptor gamma, Fc receptor beta, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, and CD66d.

[0577] In one embodiment, a CAR of the invention comprises an intracellular signalling domain of CD3-zeta.

[0578] It will be appreciated that one or more ITAMs of the intracellular signalling domain may be modified, for example by mutation. The modification may be used to increase or decrease the signalling function of the ITAM as compared with the native ITAM domain.

[0579] As mentioned above, the intracellular signalling domain may comprise a primary intracellular signalling domain by itself, or it may comprise a primary intracellular signalling domain in combination with one or more secondary intracellular signalling domains, such as one or more costimulatory signalling domains. Thus, the intracellular signalling domain of the CAR may comprise the CD3 zeta signalling domain by itself or in combination with one or more other intracellular signalling domains such as one or more costimulatory signalling domains.

[0580] The costimulatory signalling domain refers to a portion of the CAR comprising the intracellular domain of a costimulatory molecule. A costimulatory molecule may be a cell surface molecule other than an antigen receptor or its ligands that is required for an efficient response of immune cells (e.g. lymphocytes) to an antigen. Examples of such molecules include CD28, 4-1BB (CD137), 0X40, ICOS, DAP10, CD27, CD30, CD40, PD1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, and a ligand that specifically binds with CD83, and the like. For example, CD27 co-stimulation has been demonstrated to enhance expansion, effector function, and survival of human CAR T cells in vitro and augments human T cell persistence and anti-tumour activity in vivo (Song et al., Blood. 2012, 119(3):696- 706).

[0581] The intracellular signalling sequences within the intracellular portion of the CAR of the invention may be linked to each other in a random or specified o...

Claims

CLAIMS1. A binding molecule, comprising one or more polypeptide chains, said binding molecule having binding specificity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of human cytomegalovirus (HCMV), wherein ECD3 of the US28 protein comprises an amino acid sequence presented in the US28 protein at positions corresponding to positions 167 to 183 of the US28 protein encoded by human cytomegalovirus (HCMV) as set forth in SEQ ID NO: 5.

2. The binding molecule of Claim 1, wherein the binding molecule is selected from an antibody and a chimeric antigen receptor (CAR).

3. The binding molecule of Claim 1 or 2, wherein the binding molecule has binding specificity to an epitope present entirely within extracellular domain 3 (ECD3) of the US28 protein of HCMV.

4. The binding molecule of any preceding claim, wherein the binding molecule has binding specificity to a linear epitope within ECD3 of the US28 protein.

5. The binding molecule of any preceding claim, having binding specificity to an epitope within ECD3 of a US28 protein of HCMV that is HCMV strain agnostic, for example, wherein the binding molecule: (a) has a binding specificity to an epitope within ECD3 of a US28 protein of HCMV that is agnostic to two or more (such as all) of HCMV strains, (b) has a binding specificity that is agnostic to 4D-variant strains and 4N-variant strains, and / or (c) has a binding specificity that is agnostic to two or more (such as all) HCMV strains selected from the group consisting of DB, Towne, AF1, VHL / E, AD169, BL, DAVIS, JP, Merlin, PH, TB40 / E, Toledo, TR and VR1814 (FIX).

6. The binding molecule of any preceding claim, wherein the binding molecule has binding specificity to an epitope within ECD3 of the US28 protein of HCMV, irrespective of whether the ECD3 of the US28 protein comprises the sequence of a 4D-variant or a 4N-variant:- wherein the 4D-variant comprises the sequence of TKKDNQCMTDYDYLEVS (SEQ ID NO: 7) as found in ECD3 of US28 as encoded by a first group of HCMV strains, such as Towne, VR1814, TB40 / E, Merlin, JP, Ad 169, AF1, VHL / E, BL and DAVIS; and- wherein the 4N-variant comprises the sequence of TKKNNQCMTDYDYLEVS (SEQ ID NO: 6) as found in ECD3 of US28 as encoded by a second group of HCMV strains, such as Toledo, TR and DB strains.

7. The binding molecule of any preceding claim comprising one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody:(a) 1D3 as defined by SEQ ID NOs: 8, 9, 10, 14, 15 and 16, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1D3;(b) 1C10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1C10;(c) 1A10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1A10;(d) 1G9 as defined by SEQ ID NOs: 76, 77, 78, 82, 83 and 84, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1G9; and / or(e) 1E8 as defined by SEQ ID NOs: 76, 95, 96, 82, 99 and 100, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1E8; and optionally wherein the binding molecule is selected from an antibody and a CAR.

8. The binding molecule of Claim 7, comprising:(a) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable heavy chain (VH) of antibody:(I) 1D3, as defined by SEQ ID NOs: 8, 9, and 10, respectively;(ii) 1C10, as defined by SEQ ID NOs: 112, 113, and 114, respectively;(iii) 1A10, as defined by SEQ ID NOs: 112, 113, and 114, respectively;(iv) 1G9, as defined by SEQ ID NOs: 76, 77, and 78, respectively; and / or(v) 1E8, as defined by SEQ ID NOs: 76, 95, and 96, respectively; and / or(b) one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable light chain (VL) of antibody:(i) 1D3, as defined by SEQ ID NOs: 14, 15, and 16, respectively; (ii)1C10, as defined by SEQ ID NOs: 117, 83, and 118, respectively;(iii) 1A10, as defined by SEQ ID NOs: 117, 83, and 118, respectively;(iv) 1G9, as defined by SEQ ID NOs: 82, 83, and 84, respectively; and / or(v) 1E8, as defined by SEQ ID NOs: 82, 99, and 100, respectively.

9. The binding molecule of Claim 7 or 8, comprising :(a) at least one variable heavy chain (VH) polypeptide that comprises CDR, 2, and 3 sequences having the sequences of:(i) SEQ ID Nos: 8, 9, and 10, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 12;(ii) SEQ ID NOs: 112, 113, and 114, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 104;(iii) SEQ ID NOs: 112, 113, and 114, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 122;(iv) SEQ ID NOs: 76, 77, and 78, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 68; and / or(V) SEQ ID NOs: 76, 95, and 96, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 88; and / or(b) at least one variable light chain (VL) polypeptide that comprises CDR 1, and 3 sequences having the sequences of:(i) SEQ ID NOs: 14, 15, and 16, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 18;(ii) SEQ ID NOs: 117, 83, and 118, respectively, andoptionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 108;(iii) SEQ ID NOs: 117, 83, and 118, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 126;(iv) SEQ ID NOs: 82, 83, and 84, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 72; and / or(V) SEQ ID NOs: 82, 99, and 100, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 92.

10. The binding molecule of any preceding claims, wherein the binding molecule is selected from the group consisting of:(a) bivalent antibodies, such as IgG-scFv antibodies (for example, wherein a first binding domain is an intact IgG and a second binding domain is an scFv attached to the first binding domain at the N-terminus of a light chain and / or at the C-terminus of a light chain and / or at the N-terminus of a heavy chain and / or at the C-terminus of a heavy chain of the IgG, or vice versa),'(b) monovalent antibodies, such as a DuoBody® or 'knob-in-hole' bispecific antibody (for example, an scFv-KIH, scFv-KIHr, a BiTE-KIH or a BiTE-KIHr;(c) scFvz-Fc antibodies;(d) bispecific antibodies, such as bispecific T-cell engager (BiTE) antibodies;(e) dual variable domain (DVD)-Ig antibodies;(f) dual-affinity re-targeting (DART)-based antibodies (for example, DART2-Fc or DART);(g) trispecific antibodies, such as DNL-Fabs antibodies;(h) scFv-HSA-scFv antibodies;(i) single domain antibodies;(j) heavy-chain-only IgGs (hdgGs), such as camelid IgG (e.g. VHH antibodies) and shark immunoglobulin new antigen receptor (IgNAR), and single chain antibodies thereof; and(k) a chimeric antigen receptor (CAR) comprising an extracellular domain according to Claim 1 or any claim dependent thereon, for example an extracellular domain that comprises any one of options (a) to (h) of this claim, or combinations thereof.

11. The binding molecule of any preceding claim, wherein the binding molecule is a :(I) a bispecific immune cell engager antibody, for example, a bispecific T- cell engager (BiTE), and optionally wherein the BiTE antibody comprises a CD3-binding domain; or(ii) monoclonal antibody, optionally a recombinant monoclonal antibody, for example, a monoclonal antibody produced recombinantly by CHO cells.

12. A functional fragment of a binding molecule as defined by any of the preceding claims, wherein the functional fragment:(a) comprises or consists of an antigen-binding fragment of a binding molecule as defined by any of the preceding claims, or a variant, fusion or derivative thereof selected from the group consisting of: an Fv fragment (such as a single chain Fv fragment (scFv), or a disulphide-bonded Fv fragment), a Fab-like fragment (such as a Fab fragment, a Fab' fragment or a F(ab)z fragment), and single domain antibodies (dAbs, including single and dual formats, such as dAb-linker-dAb and nanobodies);(b) provides the binding characteristics as defined by any of Claims 1-6;(c) comprises the CDR sequences as defined by any one of Claims 7-9; and / or(d) comprises the VH and / or VL sequences as defined by Claim 9.

13. A binding molecule as defined by any of Claims 1-11, or a functional fragment of said binding molecule as defined by Claim 12, wherein the binding molecule or the functional fragment thereof comprises a fusion polypeptide sequence, said fusion polypeptide sequence comprising a first amino acid sequence fused a second amino acid sequence, wherein the first amino acid sequence comprises or consists of at least one of the polypeptide chains of the binding molecule or of the functional fragment thereof, and the second amino acid sequence is a fusion partner.

14. A chimeric antigen receptor (CAR) comprising:(I) an extracellular domain, wherein the extracellular domain comprises or consists of a binding molecule as defined by any of Claims 1-11 or 13, or a functional fragment of said binding molecule as defined by Claim 12 or 13;(II) a transmembrane domain; and(iii) an intracellular domain;wherein the extracellular domain of the CAR has binding specificity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of human cytomegalovirus (HCMV), and wherein ECD3 of the US28 protein comprises an amino acid sequence presented in the US28 protein at positions corresponding to positions 167 to 183 of the US28 protein encoded by human cytomegalovirus (HCMV) as set forth in SEQ ID NO: 5.

15. A CAR according to Claim 14, wherein:(a) the extracellular domain of the CAR has binding specificity to an epitope present entirely within extracellular domain 3 (ECD3) of the US28 protein of HCMV;(b) the extracellular domain of the CAR has binding specificity to a linear epitope within ECD3 of the US28 protein;(c) the extracellular domain of the CAR has binding specificity to an epitope within ECD3 of a US28 protein of HCMV that is HCMV strain agnostic, for example, binding specificity to an epitope within ECD3 of a US28 protein of HCMV that is agnostic to two or more (such as all) of HCMV strains selected from the group consisting of DB, Towne, AD169, BL, DAVIS, JP, Merlin, PH, TB40 / E, Toledo, TR, VHL / E and VR1814 (FIX); and / or(d) the extracellular domain of the CAR has specificity to an epitope within ECD3 of the US28 protein of HCMV, irrespective of whether the ECD3 of the US28 protein comprises the sequence of:- TKKDNQCMTDYDYLEVS (SEQ ID NO: 7) as found in ECD3 of US28 as encoded by a majority of HCMV strains, or- TKKNNQCMTDYDYLEVS (SEQ ID NO: 6) as found in ECD3 of US28 as encoded by a minority of HCMV strains.

16. A CAR according to Claim 14 or 15, wherein:(a) the extracellular domain of the CAR comprises one, two, three, four, five or six complementarity determining regions (CDRs) corresponding to any one, two, three, four, five or all six of the CDR sequences of antibody:(I) 1D3 as defined by SEQ ID NOs: 8, 9, 10, 14, 15 and 16, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1D3;(ii) 1C10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1C10;(Hi) 1A10 as defined by SEQ ID NOs: 112, 113, 114, 117, 83 and 118, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1A10;(iv) 1G9 as defined by SEQ ID NOs: 76, 77, 78, 82, 83 and 84, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1G9; and / or(v) 1E8 as defined by SEQ ID NOs: 76, 95, 96, 82, 99 and 100, respectively, and / or a functional variant of any one or more of said CDR sequences of antibody 1E8;(b) the extracellular domain of the CAR comprises: one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable heavy chain (VH) of antibody:(i) 1D3, as defined by SEQ ID NOs: 8, 9, and 10, respectively;(ii) 1C10, as defined by SEQ ID NOs: 112, 113, and 114, respectively;(iii) 1A10, as defined by SEQ ID NOs: 112, 113, and 114, respectively;(iv) 1G9, as defined by SEQ ID NOs: 76, 77, and 78, respectively; and / or(v) 1E8, as defined by SEQ ID NOs: 76, 95, and 96, respectively; and / or one, two, or all three, of the CDR 1, 2, and 3, sequences of the variable light chain (VL) of antibody:(vii) 1D3, as defined by SEQ ID NOs: 14, 15, and 16, respectively;(viii) 1C10, as defined by SEQ ID NOs: 117, 83, and 118, respectively;(ix) 1A10, as defined by SEQ ID NOs: 117, 83, and 118, respectively;(x) 1G9, as defined by SEQ ID NOs: 82, 83, and 84, respectively; and / or(xi) 1E8, as defined by SEQ ID NOs: 82, 99, and 100, respectively; and / or(c) the extracellular domain of the CAR comprises: at least one variable heavy chain (VH) polypeptide sequence that comprises CDR 1, 2, and 3 sequences having the sequences of:(i) SEQ ID Nos: 8, 9, and 10, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide sequence comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 12;(ii) SEQ ID NOs: 112, 113, and 114, respectively, andoptionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 104;(iii) SEQ ID NOs: 112, 113, and 114, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 122;(iv) SEQ ID NOs: 76, 77, and 78, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 68; and / or(V) SEQ ID NOs: 76, 95, and 96, respectively, and optionally wherein the at least one variable heavy chain (VH) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 88; and / or at least one variable light chain (VL) polypeptide sequence that comprises CDR 1, 2 and 3 sequences having the sequences of:(vii) SEQ ID NOs: 14, 15, and 16, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 18;(viii) SEQ ID NOs: 117, 83, and 118, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 108;(ix) SEQ ID NOs: 117, 83, and 118, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 126;(X) SEQ ID NOs: 82, 83, and 84, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 72; and / or(xi) SEQ ID NOs: 82, 99, and 100, respectively, and optionally wherein the variable light chain (VL) polypeptide comprises, consists essentially of, or consists of, the sequence of SEQ ID NO: 92.

17. A CAR according to any of Claims 14-16, wherein the extracellular domain is an antibody, for example a single-chain variable fragment (scFv).

18. A CAR according to any of Claims 14-17, wherein the transmembrane domain comprises the transmembrane domain of a protein, for example the transmembrane domain of a transmembrane receptor protein, and optionally wherein the transmembrane domain comprises the transmembrane domain of a protein selected from the group consisting of the alpha, beta or zeta chain of the T-cell receptor, CD28, CD3 epsilon, CD8, CD45 and CD4.

19. A CAR according to any of Claims 14-18, wherein the extracellular domain is connected to the transmembrane domain by a hinge region.

20. A CAR according to any of Claims 14-19, wherein the intracellular domain comprises an intracellular signalling domain, for example wherein:(a) the intracellular signalling domain comprises one or more immunoreceptor tyrosine-based activation motifs (ITAMs); and / or(b) the intracellular signalling domain comprises a signalling domain of CD3 zeta, Fc receptor gamma, Fc receptor beta, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, and CD66d.

21. A CAR according to Claim 20, wherein the intracellular domain comprises one or more costimulatory domains, for example:(a) wherein the one or more costimulatory domains includes one or more functional signalling domains obtained from a protein selected from the group consisting of CD28, 41BB, 0X40, ICOS, CD27, and DAP10;(b) wherein the intracellular domain incorporates a costimulatory domain proximal to the intracellular signalling domain,(c) wherein the intracellular domain comprises two or more costimulatory domains, for example two in-line costimulatory domains, and / or(d) wherein the intracellular domain incorporates separate cytokine signals.

22. A CAR according to any of Claims 14-21, wherein the CAR further comprises a leader sequence.

23. A nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, wherein the nucleic acid molecule comprises, or the combination of multiple distinct nucleic acid molecules collectively comprises, one or more nucleic acid sequences that, individually or in combination, encode the binding molecule of any of Claims 1-13, or the CAR of any of Claims 14-22.

24. A vector comprising a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 23.

25. A vector according to Claim 24 wherein the vector is selected from the group consisting of a retroviral vector, a plasmid, a lentivirus vector, and an adenoviral vector.

26. A cell comprising the nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 23, or the vector according to Claims 24 or 25, optionally wherein the cell expresses one or more binding molecules according to any of Claims 1-13, and / or one or more CARs according to of any of Claims 14-22, said one or more binding molecules and / or CARs being encoded by the nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 23, or the vector according to Claims 24 or 25.

27. A cell according to Claim 26, wherein the cell comprises:(a) a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 23, wherein the encoded binding molecule is an antibody according to any of Claims 1-11, a functional fragment of said antibody according to Claim 12, or an antibody of functional fragment thereof that comprises a fusion polypeptide sequence according to Claim 13; and / or(b) a vector according to Claim 25 or 26, wherein said vector comprises a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules as defined by part (a) of this claim.

28. A cell according to Claim 26, wherein the cell comprises:(a) a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 23, wherein the encoded binding molecule is a CAR according to any of Claims 14-22; and / or(b) a vector according to Claim 25 or 26, wherein said vector comprises a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules as defined by part (a) of this claim.

29. A cell comprising a binding molecule according to any of Claims 1-13 and / or a nucleic acid encoding said binding molecule, optionally wherein said nucleic acid is a nucleic acid or vector as defined by any of Claims 23 to 25.

30. The cell of Claim 29, wherein the binding molecule is an antibody according to any of Claims 1-11, a functional fragment of said antibody according to Claim 12, or an antibody of functional fragment thereof that comprises a fusion polypeptide sequence according to Claim 13, and optionally wherein the antibody is monoclonal antibody, and further for example wherein the cell is a CHO cell that recombinantly expresses the monoclonal antibody31. A cell comprising a CAR according to any of Claims 14-22 and / or a nucleic acid encoding said CAR, optionally wherein said nucleic acid is a nucleic acid or vector as defined by any of Claims 23 to 25.

32. A cell according to Claim 26, 28, 29 or 31, or a method according to Claim 32, wherein the cell is a T cell, natural killer (NK) cell, or a macrophage.

33. A cell according to Claim 31 or 32, wherein the cell is a CAR-T cell, a CAR-NK cell or a CAR-macrophage, and optionally, when the cell is a CAR-T cell, then the T-cell is selected from the group consisting of CD8+T cells, CD4+ T cells, effector T cells, helper T cells, memory T cells, cytotoxic T lymphocytes (CTLs), EBV-specific T cell receptor (TCR) or y6-T cell subtypes.

34. A method of producing a cell, the method comprising introducing a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 23, and / or a vector according to Claims 24 or 25, into a cell.

35. A method of producing a binding molecule according to any of Claims 1 to 13, or a CAR according to any of Claims 14 to 22, the method comprising: expressing a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 23, and / or a vector according to Claims 24 or 25, in a cell.

36. The method of Claim 35, comprising the step of isolating the thus- produced binding molecule from the cell; and optionally, wherein the binding molecule is an antibody according to any of Claims 1-11, a functional fragment of said antibody according to Claim 12, or an antibody of functional fragment thereof that comprises a fusion polypeptide sequence according to Claim 13.27037. An isolated binding molecule that is obtained, or obtainable, by the method of Claim 36, optionally, wherein the isolated binding molecule is further formulated for administration to a subject.

38. A conjugate, the conjugate comprising a moiety conjugated to a binding molecule as defined by any of Claims 1-11 or 13, or to a functional fragment of said binding molecule as defined by Claim 12 or 13.

39. The conjugate of Claim 38, wherein said moiety is a therapeutic, prophylactic, diagnostic, prognostic, or theragnostic moiety.

40. The conjugate of Claim 38 or 39, wherein said moiety is a drug (for example, wherein the conjugate is an antibody-drug conjugate ("ADC")) and / or a radioactive moiety (for example, wherein the conjugate is suitable for use in radioimmunotherapy ("RIT")).

41. A method of producing a conjugate according to any of Claims 38 to 40, the method comprising the steps of:(a) providing a binding molecule as defined by any of Claims 1-11 or 13, or a functional fragment of said binding molecule as defined by Claim 12 or 13; and(b) conjugating a moiety to the binding molecule as defined by any of Claims 1-11 or 13, or to the functional fragment of said binding molecule as defined by Claim 12 or 13.

42. The method of Claim 41, comprising the step of isolating the thus- produced conjugate.

43. The conjugate of any of Claims 38 to 40, or the method of Claim 41 or 42, wherein the binding molecule is an antibody according to any of Claims 1-11, a functional fragment of said antibody according to Claim 12, or an antibody of functional fragment thereof that comprises a fusion polypeptide sequence according to Claim 13.

44. An isolated conjugate that is obtained, or obtainable, by the method of Claim 42 or 43, optionally, wherein the isolated conjugate is further formulated for administration to a subject.27145. A method of combating HCMV or a disease or condition associated with HCMV, the method comprising administering to a subject, or to ex vivo or in vitro cellular material, any one or more agents selected from the group consisting of: i. a binding molecule according to any of Claims 1-11 or 13, ii. a functional fragment of said binding molecule as defined by Claim 12 or 13, ill. an isolated binding molecule according to Claim 37, iv. a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 23, v. a vector according to Claims 24 or 25, vi. a cell according to any of Claims 26 to 33, vii. a conjugate according to any of Claims 38 to 40, and viii. an isolated conjugate according to Claim 44.

46. One or more agents for use in combating a disease or condition associated with HCMV in a subject, or in ex vivo or in vitro cellular material, wherein the one or more agents is, or are, each individually selected from the group consisting of: i. a binding molecule according to any of Claims 1-11 or 13,II. a functional fragment of said binding molecule as defined by Claim 12 or13, ill. an isolated binding molecule according to Claim 37, iv. a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 23, v. a vector according to Claims 24 or 25, vi. a cell according to any of Claims 26 to 33, vii. a conjugate according to any of Claims 38 to 40, and viii. an isolated conjugate according to Claim 44.

47. Use one or more agents in the manufacture of a medicament for combating a disease or condition associated with HCMV in a subject, or in ex vivo or in vitro cellular material, wherein the one or more agents is, or are, each individually selected from the group consisting of: i. a binding molecule according to any of Claims 1-11 or 13,II. a functional fragment of said binding molecule as defined by Claim 12 or13, ill. an isolated binding molecule according to Claim 37,272iv. a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 23, v. a vector according to Claims 24 or 25, vi. a cell according to any of Claims 26 to 33, vii. a conjugate according to any of Claims 38 to 40, and viii. an isolated conjugate according to Claim 44.

48. A method according to Claim 45, the one or more agents for use according to Claim 46, or the use according to Claim of 47, wherein:I. the disease or condition is an HCMV infection or is associated with an HCMV infection, and optionally the HCMV infection comprises a multi-strain HCMV infection, wherein the multi-strain HCMV infection comprises infection with more than one different strain of HCMV, for example one or more HCMV strain that encodes the 4N-variant of ECD3 of US28 and one or more HCMV strain that encodes the 4D-variant of ECD3 of US28; ii. the disease or condition is a latent HCMV infection (optionally a multistrain latent HCMV infection) or is associated with a latent HCMV infection (optionally a multi-strain latent HCMV infection); ill. the disease or condition is a lytic HCMV infection (optionally a multistrain lytic HCMV infection) or is associated with a lytic HCMV infection (optionally a multi-strain lytic HCMV infection); iv. the disease or condition is a congenital single or multi-strain HCMV infection, such as a latent congenital single or multi-strain HCMV infection or a lytic congenital single or multi-strain HCMV infection; v. the disease or condition is cancer; vi. the disease or condition is a HCMV-infected cancer (optionally a multistrain HCMV infected cancer), such as latent HCMV-infected cancer (optionally a multistrain latent HCMV infected cancer); vii. the disease or condition is an epithelial cancer; optionally wherein the epithelial cancer is breast cancer; for example, wherein the breast cancer is triple negative breast cancer (TNBC), or a HER2-postive breast cancer (such as a triple positive breast cancer "TPBC"); viii. the disease is a metastasising and / or aggressive form of cancer, such as a metastasising and / or aggressive form of HCMV-infected cancer (e.g. a cancer having a latent HCMV infection and / or a multi-strain HCMV infection); ix. the disease or condition is not glioblastoma; x. the subject in need thereof has been diagnosed with HCMV-infected cancer cells, such as latent HCMV-infected cancer cells or cancer cells with a productive273HCMV infection (including but not limited to tumour associated macrophages with productive HCMV infection); and / or xi. the subject in need thereof is, or is intended to be, the recipient of an organ donation, or an organ donor.

49. A method according to Claim 45 or 48, the one or more agents for use according to Claim 46 or 48, or the use according to Claim of 47 or 48, wherein the agent is selected from the group consisting of: i. a therapeutic antibody as defined by any of Claims 1 to 11, ii. a functional fragment of said therapeutic antibody as defined by Claim 12, iii. a therapeutic antibody that comprises as fusion polypeptide sequence as defined by Claim 13; and iv. a functional fragment of said therapeutic antibody that comprises as fusion polypeptide sequence as defined by Claim 13.

50. A method according to Claim 45 or 48, the one or more agents for use according to Claim 46 or 48, or the use according to Claim of 47 or 48, wherein the agent is a bispecific antibody, for example a bispecific immune cell engager antibody, such as bispecific T-cell engager (BITE) antibody, optionally wherein the BITE antibody comprises a CD3-binding domain.

51. A method according to Claim 45 or 48, the one or more agents for use according to Claim 46 or 48, or the use according to Claim of 47 or 48, wherein the agent is a conjugate according to any of Claims 38 to 40, such as conjugate that is an antibody-drug conjugate ("ADC"), or a conjugate that comprises radioactive moiety, such as conjugate that is suitable for use in radioimmunotherapy ("RIT").

52. A method according to Claim 45 or 48, the one or more agents for use according to Claim 46 or 48, or the use according to Claim of 47 or 48, wherein the agent is a cell comprising a CAR according to any of Claims 31 to 33, for example a CAR-T cell, a CAR-NK cell or a CAR-macrophage.

53. A method according to Claim 45, or any of Claims 48 to 52, the one or more agents for use according to Claim 46 or any of Claims 48 to 52, or the use according to any of Claims 47 to 52,274wherein the subject is one who is administered a further substance, such as a further therapeutic, prophylactic, diagnostic, prognostic, or theragnostic substance, and optionally wherein the further substance is administered separately, sequentially or simultaneously with the, or each of the one or more agents.

54. An agent for use in medicine, wherein the agent is selected from the group consisting: i. a binding molecule according to any of Claims 1-11 or 13, ii. a functional fragment of said binding molecule as defined by Claim 12 or 13, ill. an isolated binding molecule according to Claim 37, iv. a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 23, v. a vector according to Claims 24 or 25, vi. a cell according to any of Claims 26 to 33, vii. a conjugate according to any of Claims 38 to 40, and viii. an isolated conjugate according to Claim 44.

55. A vaccine composition suitable for use in vaccinating against, reducing the risk of, preventing, or combating a disease or condition associated with human cytomegalovirus (HCMV), wherein the vaccine is an active or passive vaccine, wherein the vaccine triggers and / or provides an immune response directed to an epitope present within ECD3 of a US28 protein of HCMV, and wherein ECD3 of the US28 protein comprises an amino acid sequence presented in the US28 protein at positions corresponding to positions 167 to 183 of the US28 protein encoded by HCMV as set forth in SEQ ID NO: 5.

56. The vaccine composition of Claim 55, wherein the vaccine triggers and / or provides an immune response:(a) to one or more epitopes present entirely within extracellular domain 3 (ECD3) of the US28 protein of HCMV;(b) to one or more linear epitopes within ECD3 of the US28 protein;(c) to one or more epitopes within ECD3 of a US28 protein of HCMV that is an epitope, or are epitopes, present in identical form in both the 4D-variant strains and 4N-variant strains of HCMV, wherein the 4D-variant strain of HCMV encodes a US28 protein comprising an ECD3 having the sequence of TKKDNQCMTDYDYLEVS (SEQ ID275NO:7) and wherein the 4N-variant strain of HCMV encodes a US28 protein comprising an ECD3 having the sequence of TKKNNQCMTDYDYLEVS (SEQ ID NO:6); and / or(d) wherein the immune response that is triggered or provided by the vaccine is HCMV strain agnostic to the 4D-variant strains and 4N-variant strains of HCMV, and triggers and / or provides an immune response that is directed to one or more of the 4D-variant HCMV strains selected from Towne, VR1814, TB40 / E, Merlin, JP, Ad 169, VHL / E, AF1, BL and DAVIS and is also directed to one or more of the 4N-variant HCMV strains selected from Toledo, TR and DB.

57. The vaccine composition of Claim 55 or 56, wherein:(I) the vaccine is a passive vaccine, and / or optionally comprises:(a) one or more binding molecules according to any of Claims 1-11 or 13,(b) one or more functional fragments of said one or more binding molecules as defined by Claim 12 or 13,(c) one or more isolated binding molecules according to Claim 37,(d) one or more nucleic acid molecules, or combination of multiple distinct nucleic acid molecules, according to Claim 23,(e) one or more vectors according to Claims 24 or 25,(f) one or more cells according to any of Claims 26 to 33,(g) one or more conjugates according to any of Claims 38 to 40, and / or(h) one or more isolated conjugates according to Claim 44; and / or(ii) the vaccine is an active vaccine, and / or optionally comprises:(a) one or more peptides or polypeptides according to any of Claims 84 to 88, a combination of at least two distinct peptides and / or polypeptides according to Claim 89, a fusion protein according to Claim 90, a combination of at least two distinct fusion proteins according to Claim 91, a conjugate according to any of Claims 92 to 94, and / or a combination of at least two distinct conjugates according to Claim 95; and / or(b) one or more nucleic acid molecules, or combination of multiple distinct nucleic acid molecules, according to Claim 101 or 102, or the vector according to Claims 103 or 104; and / or(c) a dendritic cell, or a homogeneous or heterogeneous population thereof, wherein the or each dendritic cells is loaded with one or more of the following: a peptide or polypeptide according to any of Claims 84 to 88, a combination of at least two distinct peptides and / or polypeptides according to Claim 89, a fusion protein276according to Claim 90, a combination of at least two distinct fusion proteins according to Claim 91, a conjugate according to any of Claims 92 to 94, a combination of at least two distinct conjugates according to Claim 95, a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 101 or 102, and / or one or more vector according to Claims 103 or 104.

58. A method of vaccinating against, reducing the risk of, preventing, and / or combating a disease or condition associated with HCMV, the method comprising administering to a subject a vaccine according to any of Claims 55 to 57, optionally wherein the disease or condition is a latent HCMV infection, or is a disease or condition associated with a latent HCMV infection.

59. A vaccine according to any of Claims 55 to 57 for use in vaccinating against, reducing the risk of, preventing, and / or combating a disease or condition associated with HCMV in a subject, optionally wherein the disease or condition is a latent HCMV infection, or is a disease or condition associated with a latent HCMV infection.

60. Use of a vaccine according to any of Claims 55 to 57 in the manufacture of a medicament for vaccinating against, reducing the risk of, preventing, and / or combating a disease or condition associated with HCMV in a subject, optionally wherein the disease or condition is a latent HCMV infection, or is a disease or condition associated with a latent HCMV infection.

61. The vaccine composition of any of Claims 55 to 57, the method of Claim 58, the vaccine composition for use according to Claim 59, or the use according to Claim of 60, wherein:I. the disease or condition is an HCMV infection or is associated with an HCMV infection, and optionally the HCMV infection comprises a multi-strain HCMV infection, wherein the multi-strain HCMV infection comprises infection with more than one different strain of HCMV, for example one or more HCMV strain that encodes the 4N-variant of ECD3 of US28 and one or more HCMV strain that encodes the 4D-variant of ECD3 of US28; ii. the disease or condition is a latent HCMV infection (optionally a multistrain latent HCMV infection) or is associated with a latent HCMV infection (optionally a multi-strain latent HCMV infection);277Hi. the disease or condition is a lytic HCMV infection (optionally a multistrain lytic HCMV infection) or is associated with a lytic HCMV infection (optionally a multi-strain lytic HCMV infection); iv. the disease or condition is a congenital single or multi-strain HCMV infection, such as a latent congenital single or multi-strain HCMV infection or a lytic congenital single or multi-strain HCMV infection; v. the disease or condition is cancer; vi. the disease or condition is a HCMV-infected cancer (optionally a multistrain HCMV infected cancer), such as latent HCMV-infected cancer (optionally a multistrain latent HCMV infected cancer); vii. the disease or condition is an epithelial cancer; optionally wherein the epithelial cancer is breast cancer; for example, wherein the breast cancer is triple negative breast cancer (TNBC), or a HER2-postive breast cancer (such as a triple positive breast cancer "TPBC"); viii. the disease or condition is a metastasising and / or aggressive form of cancer, such as a metastasising and / or aggressive form of HCMV-infected cancer (e.g. a cancer having a latent HCMV infection and / or a multi-strain HCMV infection); ix. the disease or condition is not glioblastoma; x. the subject in need thereof has been diagnosed with HCMV-infected cancer cells, such as latent HCMV-infected cancer cells or cancer cells with a productive HCMV infection (including but not limited to tumour associated macrophages with productive HCMV infection); and / or xi. the subject in need thereof is, or is intended to be, the recipient of an organ donation, or an organ donor.

62. A method of assessing one or more biological conditions and / or biological characteristics of a subject and / or of ex vivo biological material, wherein the method comprises:(a) contacting the subject and / or the ex vivo biological material with a binding molecule as defined by any of Claims 1 to 13, or a conjugate as defined by any of Claims 38 to 40, 43 or 44; and(b) making an assessment of the subject and / or the ex vivo biological material based on a direct and / or indirect measurement of the binding of the binding molecule or conjugate to the subject and / or the ex vivo biological material.

63. The method of Claim 62, wherein the method comprises an ELISA method, and optionally the method is performed on ex vivo biological material, such as one or more body fluids.27864. The method of Claim 62, wherein the method comprises a flow cytometry method, and optionally the method is a method for assessing an ex vivo blood sample and / or ex vivo bone marrow sample from a subject.

65. The method of Claim 62, wherein the method comprises the use of a conjugate as defined by any of Claims 38 to 40, 43 or 44, wherein the conjugate comprises a detectable moiety, such as radioactive moiety, and the method comprises detection of the radioactive moiety in the subject and / or the ex vivo biological material, for example, wherein the method is a method of immune-positron emitting (PET) imaging.

66. The method of Claim 62, wherein the method is a method of immunohistochemistry, performed on a sample of ex vivo biological material.

67. The method of any of Claims 62 to 66, wherein the method is performed on a subject, or on ex vivo biological material obtained from the subject, for the purposes of making a diagnostic or prognostic assessment of a disease or condition associated with HCMV in the subject.

68. The method of Claim 67, wherein:I. the disease or condition is an HCMV infection or is associated with an HCMV infection, and optionally the HCMV infection comprises a multi-strain HCMV infection, wherein the multi-strain HCMV infection comprises infection with more than one different strain of HCMV, for example one or more HCMV strain that encodes the 4N-variant of ECD3 of US28 and one or more HCMV strain that encodes the 4D-variant of ECD3 of US28; ii. the disease or condition is a latent HCMV infection (optionally a multistrain latent HCMV infection) or is associated with a latent HCMV infection (optionally a multi-strain latent HCMV infection); ill. the disease or condition is a lytic HCMV infection (optionally a multistrain lytic HCMV infection) or is associated with a lytic HCMV infection (optionally a multi-strain lytic HCMV infection); iv. the disease or condition is a congenital single or multi-strain HCMV infection, such as a latent congenital single or multi-strain HCMV infection or a lytic congenital single or multi-strain HCMV infection; v. the disease or condition is cancer;279vi. the disease or condition is a HCMV-infected cancer (optionally a multistrain HCMV infected cancer), such as latent HCMV-infected cancer (optionally a multistrain latent HCMV infected cancer); vii. the disease or condition is an epithelial cancer; optionally wherein the epithelial cancer is breast cancer; for example, wherein the breast cancer is triple negative breast cancer (TNBC), or a HER2-postive breast cancer (such as a triple positive breast cancer "TPBC"); viii. the disease or condition is a metastasising and / or aggressive form of cancer, such as a metastasising and / or aggressive form of HCMV-infected cancer (e.g. a cancer having a latent HCMV infection and / or a multi-strain HCMV infection); ix. the disease or condition is not glioblastoma; and / or x. the subject in need thereof has been diagnosed with HCMV-infected cancer cells, such as latent HCMV-infected cancer cells or cancer cells with a productive HCMV infection (including but not limited to tumour associated macrophages with productive HCMV infection), and / or multi-strain HCMV infected cancer cells; xi. the subject is, or is intended to be, the recipient of an organ donation, or an organ donor.

69. A method of combating a HCMV infection (such as a latent HCMV infection and / or a lytic HCMV infection and / or a multi-strain HCMV infection) in living ex vivo biological material, the method comprising contacting the living ex vivo biological material with any one or more agents selected from the group consisting of: i. a binding molecule according to any of Claims 1-11 or 13,II. a functional fragment of said binding molecule as defined by Claim 12 or13, ill. an isolated binding molecule according to Claim 37, iv. a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 23, v. a vector according to Claims 24 or 25, vi. a cell according to any of Claims 26 to 33, vii. a conjugate according to any of Claims 38 to 40, and viii. an isolated conjugate according to Claim 44.

70. Living ex vivo biological material that is obtained, or obtainable, by the method of Claim 69.

71. The method of Claim 69 , or living ex vivo biological material according to Claim 70, wherein the ex vivo living biological material comprises, consists280essentially of, or consists of, living ex vivo biological material selected from the group that includes: one or more types of ex vivo cells; one or more types of ex vivo cell cultures; one or more types of ex vivo tissues; one or more types of ex vivo tissue cultures; one or more types of ex vivo organoids; one or more types of ex vivo organoid cultures; one or more types of ex vivo organs; and / or one or more types of ex vivo organ cultures.

72. A method of treating a subject in need thereof, comprising administering ex vivo living biological material as defined by Claim 60 or 61, to the subject.

73. The method of Claim 72, wherein the method is a method of transplantation of the ex vivo living biological material, such as an organ or tissue transplant.

74. A method of screening for a binding molecule having binding specificity to an epitope within extracellular domain 3 (ECD3) of a US28 protein of human cytomegalovirus (HCMV), wherein ECD3 of the US28 protein comprises an amino acid sequence presented in the US28 protein at positions corresponding to positions 167 to 183 of the US28 protein encoded by HCMV as set forth in SEQ ID NO: 5, the method comprising:(a) providing one or more peptides corresponding an amino acid sequence present in ECD3 of the US28 protein, optionally wherein the one or more peptides are provided in the form of one or more peptides or polypeptides according to any of Claims 84 to 88, a combination of at least two distinct peptides and / or polypeptides according to Claim 89, one or more fusion proteins according to Claim 90, a combination of at least two distinct fusion proteins according to Claim 91, a conjugate according to any of Claims 92 to 94, or a combination of at least two distinct conjugates according to Claim 95;(b) providing one or more candidate binding molecules;(c) determining the binding specificity and / or binding affinity of one or more candidate binding molecules to the one or more peptides.

75. The method of Claim 74, wherein the, or each, of the one or more peptides comprise, consist essentially of, or consist of, the polypeptide sequence TKKDNQCMTDYDYLEVS (SEQ ID NO: 7) and / or TKKNNQCMTDYDYLEVS (SEQ ID NO: 6), and / or an immunogenic fragment of either or both;281optionally wherein the one or more peptides are provided in the form of one or more peptides or polypeptides according to any of Claims 84 to 88, a combination of at least two distinct peptides and / or polypeptides according to Claim 89, one or more fusion proteins according to Claim 90, a combination of at least two distinct fusion proteins according to Claim 91, a conjugate according to any of Claims 92 to 94, or a combination of at least two distinct conjugates according to Claim 95.

76. The method of Claim 74 or 75, wherein the one or more candidate binding molecules are antibodies and / or CARs, such as any one or more of:(a) bivalent antibodies, such as IgG-scFv antibodies (for example, wherein a first binding domain is an intact IgG and a second binding domain is an scFv attached to the first binding domain at the N-terminus of a light chain and / or at the C-terminus of a light chain and / or at the N-terminus of a heavy chain and / or at the C-terminus of a heavy chain of the IgG, or vice versa),'(b) monovalent antibodies, such as a DuoBody® or 'knob-in-hole' bispecific antibody (for example, an scFv-KIH, scFv-KIHr, a BiTE-KIH or a BiTE-KIHr;(c) scFvz-Fc antibodies;(d) bispecific antibodies, such as bispecific T-cell engager (BiTE) antibodies;(e) dual variable domain (DVD)-Ig antibodies;(f) dual-affinity re-targeting (DART)-based antibodies (for example, DART2- Fc or DART);(g) trispecific antibodies, such as DNL-Fabs antibodies;(h) scFv-HSA-scFv antibodies; and(i) a chimeric antigen receptor (CAR).

77. The method of any of Claims 74 to 76, further comprising the step of selecting a candidate binding molecule based on the binding specificity and / or binding affinity to the one or more peptides.

78. The method of Claim 77, wherein the selected candidate binding molecule:(a) has binding specificity to an epitope present entirely within extracellular domain 3 (ECD3) of the US28 protein of HCMV;(b) has binding specificity to a linear epitope within ECD3 of the US28 protein;(c) has binding specificity to an epitope within ECD3 of a US28 protein of HCMV that is HCMV strain agnostic, for example, binding specificity to an epitope within ECD3 of a US28 protein of HCMV that is agnostic to two or more (such as all) of HCMV282strains selected from the group consisting of DB, Towne, AD169, DAVIS, BL, JP, Merlin, PH, TB40 / E, Toledo, VHL / E, TR and VR1814 (FIX); and / or(d) has specificity to an epitope within ECD3 of the US28 protein of HCMV, irrespective of whether the ECD3 of the US28 protein comprises the sequence of TKKDNQCMTDYDYLEVS (SEQ ID NO:7) or TKKNNQCMTDYDYLEVS (SEQ ID NO: 6).

79. The method of Claim 77 or 78, wherein the selected candidate binding molecule has a binding specificity and / or binding affinity to the one or more peptides that is equivalent to the binding specificity and / or binding affinity to the same one or more peptides as demonstrated by an antibody that comprises a variable heavy chain (VH) polypeptide that consists of the sequence of SEQ ID NO: 12 and a variable light chain (VL) polypeptide that consists of the sequence of SEQ ID NO: 18.

80. A method of producing a composition that comprises multiple copies of a binding molecule, said method comprising causing the reproduction of a selected candidate binding molecule that has been selected in accordance with the method of any of Claims 77, 78 or 79.

81. A method of assessing a selected candidate binding molecule that has been selected in accordance with the method of any of Claims 77, 78 or 79, said method comprising and identifying the structure(s) within the selected candidate binding molecule that provides its binding characteristics, for example, identifying the, or each, CDR sequence in a selected candidate binding molecule that is an antibody or CAR.

82. A method of producing a composition that comprises multiple copies of a binding molecule, wherein said binding molecule comprises the, or each, of the structure(s) that have been identified within a selected candidate binding molecule as providing its binding characteristics, in accordance with the method of Claim 81, said method comprising causing the reproduction of the binding molecule.

83. A composition of binding molecule obtained by the method of Claim 80 or 82.

84. A peptide or polypeptide comprising, consisting essentially of, or consisting of, the sequence TKKNNQCMTDYDYLEVS (SEQ ID NO: 6), or comprising the sequence of an immunogenic fragment of SEQ ID NO: 6, wherein said peptide or polypeptide is not the US28 protein, and preferably wherein the only US28-derived283sequence in said peptide or polypeptide is the sequence of SEQ ID NO: 6 or the sequence of the immunogenic fragment of SEQ ID NO: 6.

85. A peptide or polypeptide comprising, consisting essentially of, or consisting of, the sequence TKKDNQCMTDYDYLEVS (SEQ ID NO:7), or comprising the sequence of an immunogenic fragment of SEQ ID NO: 7, wherein said peptide or polypeptide is not the US28 protein, and preferably wherein the only US28-derived sequence in said peptide or polypeptide is the sequence of SEQ ID NO: 7 or the sequence of the immunogenic fragment of SEQ ID NO: 7.

86. The peptide or polypeptide of Claim 84 or 85, wherein the immunogenic fragment of the reference sequence SEQ ID NO: 6 or 7 comprises less than the full sequence of the reference sequence, and comprises at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 consecutive amino acids of the reference sequence.

87. The peptide or polypeptide of Claim 84, 85 or 86, wherein the immunogenic fragment comprises, consists essentially of, or consists of, a sequence that is common to, and present within, both of SEQ ID NO: 6 and SEQ ID NO:7.

88. A peptide or polypeptide comprising, consisting, or consisting essentially of, the sequence of an immunogenic fragment or variant of TKKNNQCMTDYDYLEVS (SEQ ID NO: 6) or TKKDNQCMTDYDYLEVS (SEQ ID NO: 7), wherein the immunogenic fragment or variant comprises the sequence of the epitope within ECD3 of US28 that is bound by antibody 1D3, 1C10, 1A10, 1G9, and / or 1E8.

89. A combination of at least two distinct peptides and / or polypeptides, comprising a first peptide or polypeptide and a second peptide or polypeptide, wherein: the first peptide or polypeptide comprises a comprises, consists essentially of, or consists of, the sequence TKKNNQCMTDYDYLEVS (SEQ ID NO: 6), or an immunogenic fragment thereof, such as an immunogenic fragment as defined by Claim 86 or 88, with the proviso that said immunogenic fragment comprises at least the 4N amino acid of SEQ ID NO: 6; and the second peptide or polypeptide comprises a comprises, consists essentially of, or consists of, the sequence TKKDNQCMTDYDYLEVS (SEQ ID NO: 7), or an immunogenic fragment thereof, such as an immunogenic fragment as defined by Claim 86 or 88, with the proviso that said immunogenic fragment comprises at least the 4D amino acid of SEQ ID NO: 7.28490. A fusion protein comprising, consisting essentially of, or consisting of, a first amino acid sequence fused, either directly or via one or more linker amino acid sequences, to a second amino acid sequence, wherein the first amino acid sequence is the sequence of a peptide or polypeptide as defined by any of Claims 84-88; and the second amino acid sequence is a fusion partner, optionally wherein the fusion partner is a carrier protein, such as a carrier protein that is selected to provide a fusion protein that is suitable for immunisation and generation of antibodies against the first amino acid sequence, optionally wherein the carrier protein is selected from the group consisting of keyhole limpet hemocyanin (KLH), HSA (human serum albumin), BSA (bovine serum albumin), OVA (ovalbumin), tetanus toxoid (TT), diphtheria toxoid (DT), a genetically modified cross-reacting material (CRM) of diphtheria toxin, meningococcal outer membrane protein complex (OMPC) and H. influenzae protein D (HiD).

91. A combination of at least two distinct fusion proteins, comprising a first fusion protein, and a second fusion protein, wherein: the first fusion protein according to Claim 90 comprises, as the first amino acid sequence of the first fusion protein, a sequence that comprises a comprises, consists essentially of, or consists of, the sequence TKKNNQCMTDYDYLEVS (SEQ ID NO: 6), or an immunogenic fragment thereof, such as an immunogenic fragment as defined by Claim 86 or 88, with the proviso that said immunogenic fragment comprises at least the 4N amino acid of SEQ ID NO:6; and the second fusion protein according to Claim 90 comprises, as the first amino acid sequence of the second fusion protein, a sequence that comprises, consists essentially of, or consists of, the sequence TKKDNQCMTDYDYLEVS (SEQ ID NO: 7), or an immunogenic fragment thereof, such as an immunogenic fragment as defined by Claim 86 or 88, with the proviso that said immunogenic fragment comprises at least the 4D amino acid of SEQ ID NO:7.

92. A conjugate, comprising a moiety conjugated to a peptide or polypeptide as defined by any of Claims 84-88, or to a fusion protein as defined by Claim 90.

93. The conjugate of Claim 92, wherein:(i) the moiety conjugated directly to the peptide or polypeptide as defined by any of Claims 84-88, or to the fusion protein as defined by Claim 90; or285(ii) the moiety conjugated indirectly, such as via a linker, to the peptide or polypeptide as defined by any of Claims 84-88, or to the fusion protein as defined by Claim 90.

94. The conjugate of Claim 92 or 93, wherein said moiety is a carrier, for example a carrier protein, for example a carrier selected from KLH (keyhole limpet hemocyanin), HSA (human serum albumin), BSA (bovine serum albumin), OVA (ovalbumin), tetanus toxoid (TT), diphtheria toxoid (DT), a genetically modified crossreacting material (CRM) of diphtheria toxin, meningococcal outer membrane protein complex (OMPC) and H. influenzae protein D (HiD).

95. A combination of at least two distinct conjugates, wherein the combination comprises: a first conjugate according to any of Claims 92-94, wherein the first conjugate comprises, consists essentially of, or consists of, a moiety conjugated to a peptide or polypeptide, wherein the peptide or polypeptide comprises, consists essentially of, or consists of, the sequence TKKNNQCMTDYDYLEVS (SEQ ID NO: 6), or an immunogenic fragment thereof, such as an immunogenic fragment as defined by Claim 86 or 88, with the proviso that said immunogenic fragment comprises at least the 4N amino acid of SEQ ID NO:6; and a second conjugate according to any of Claims 92-94, wherein the second conjugate comprises, consists essentially of, or consists of, a moiety conjugated to a peptide or polypeptide, wherein the peptide or polypeptide comprises, consists essentially of, or consists of, the sequence TKKDNQCMTDYDYLEVS (SEQ ID NO: 7), or an immunogenic fragment thereof, such as an immunogenic fragment as defined by Claim 86 or 88, with the proviso that said immunogenic fragment comprises at least the 4D amino acid of SEQ ID NO:7.

96. A method of producing a conjugate according to any of Claims 92-94, the method comprising the steps of:(a) providing a peptide or polypeptide as defined by any of Claims 84-88, or a fusion protein as defined by Claim 90; and(b) conjugating a moiety to the peptide or polypeptide as defined by any of Claims 84-88, or to the fusion protein as defined by Claim 90.

97. A method of producing a combination of at least two distinct conjugates as defined to Claim 95, the method comprising the steps of:286(a) producing the first conjugate, as defined by Claim 90, by a method according to Claim 95;(b) producing the second conjugate, as defined by Claim 90, by a method according to Claim 95; and(c) combining the first and second conjugates, thereby to form a combination according to Claim 95.

98. A method of producing a combination of at least two distinct conjugates as defined to Claim 95, the method comprising the steps of:(a) providing a combination of at least two distinct peptides and / or polypeptides according to Claim 89, or a combination of at least two distinct fusion proteins according to Claim 91; and(b) conjugating a moiety to the combination of at least two distinct peptides and / or polypeptides according to Claim 89, or to the combination of at least two distinct fusion proteins according to Claim 91, thereby to form a combination according to Claim 95.

99. The method of Claim 96, 97 or 98, comprising the step of isolating the thus-produced conjugate or combination of conjugates.

100. An isolated conjugate, or combination of conjugates, that is obtained, or obtainable, by the method of any of Claims 97-99, optionally, wherein the isolated conjugate is further formulated for administration to a subject.

101. A nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, wherein the nucleic acid molecule comprises, or the combination of multiple distinct nucleic acid molecules collectively comprises, one or more nucleic acid sequences that, individually or in combination, encode one or more peptides and / or polypeptides according to any of Claims 84 to 88, a combination of at least two distinct peptides and / or polypeptides according to Claim 89, a fusion protein according to Claim 90, and / or a combination of at least two distinct fusion proteins according to Claim 91.

102. The nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 101, wherein the, or each, nucleic acid molecule is independently selected from a DNA or RNA molecule.

103. A vector comprising a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 101 or 102.

104. A vector according to Claim 103 wherein the vector is selected from the group consisting of a retroviral vector, a plasmid, a lentivirus vector, and an adenoviral vector.

105. A cell comprising the nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 101 or 102, or the vector according to Claims 103 or 104, optionally wherein the cell expresses one or more peptide or polypeptide according to any of Claims 84 to 88, a combination of at least two distinct peptides and / or polypeptides according to Claim 89, a fusion protein according to Claim 90, or a combination of at least two distinct fusion proteins according to Claim 91.

106. A cell that is exposed to, and / or comprising, a one or more peptide or polypeptide according to any of Claims 84 to 88, a combination of at least two distinct peptides and / or polypeptides according to Claim 89, a fusion protein according to Claim 90, a combination of at least two distinct fusion proteins according to Claim 91, a conjugate according to any of Claims 92 to 94, a combination of at least two distinct conjugates according to Claim 95, a nucleic acid molecule, or combination of multiple distinct nucleic acid molecules, according to Claim 101 or 102, or a vector according to Claims 103 or 104.

107. A method of isolating and / or enriching cells comprising a T cell receptor (TCR) with specificity to an epitope in ECD3 of US28 (e.g. naturally occurring T cells, or recombinant cells expressing a CAR according to of any of Claims 14-22, for example CAR T-cells, CAR NK-cells and / or CAR-macrophages), wherein the method comprises the step of using an agent, wherein the agent is selected from the group consisting of a peptide or polypeptide according to any of Claims 84 to 88, a combination of at least two distinct peptides and / or polypeptides according to Claim 89, a fusion protein according to Claim 90, a combination of at least two distinct fusion proteins according to Claim 91, a conjugate according to any of Claims 92 to 94, and / or a combination of at least two distinct conjugates according to Claim 95, to isolate and / or enrich cells with binding specificity to one or both of the sequences of SEQ ID Nos: 6 and / or 7.

108. The method of Claim 107, wherein the sequences are formulated as an MHC tetramer, for example a Class I MHC tetramer for antigen-specific CD8+T cells detection, a Class II MHC tetramer for antigen-specific CD4+T cells detection, a fluorophore- labelled tetramer for flow cytometry or fluorescence microscopy.

109. The method of Claim 107 or 108, wherein the T-cell is selected from the group consisting of CD8+T cells, CD4+ T cells, effector T cells, helper T cells, memory T cells, cytotoxic T lymphocytes (CTLs), EBV-specific T cell receptor (TCR) or y6-T cell subtypes.

110. An MHC tetramer comprising a peptide or polypeptide according to any of Claims 84 to 88, a combination of at least two distinct peptides and / or polypeptides according to Claim 89, optionally wherein the peptide comprises or corresponds to SEQ ID NO:6 or SEQ ID NO:7, or an immunogenic fragment of either or both, for example wherein the MHC tetramer is a Class I MHC tetramer for antigenspecific CD8+T cells detection, a Class II MHC tetramer for antigen-specific CD4+T cells detection, a fluorophore-labelled tetramer for flow cytometry or fluorescence microscopy.

111. The MHC tetramer of Claim 110 for use in isolating and / or enriching cells comprising a T cell receptor (TCR) with specificity to an epitope in ECD3 of US28, for example, a T-cell selected from the group consisting of CD8+T cells, CD4+T cells, effector T cells, helper T cells, memory T cells, cytotoxic T lymphocytes (CTLs), EBV- specific T cell receptor (TCR) or y6-T cell subtypes and / or a recombinant cell expressing a CAR according to of any of Claims 14-22, for example a CAR T-cell, CAR NK-cell and / or CAR-macrophage.289