Monoclonal Anti-PTK7 antibodies and derivatives

Abstract

The present invention relates to a novel anti-PTK7, protein tyrosine kinase 7, antibody and derivatives thereof. The antibody and / or derivatives can be used for therapy and / or imaging, diagnosis and / or immunostaining.

Description

[0001] MONOCLONAL ANTI-PTK7 ANTIBODIES AND DERIVATIVES

[0002] FIELD

[0003] The present invention relates to a novel anti-PTK7, protein tyrosine kinase 7, antibody and derivatives thereof. The antibody and / or derivatives can be used for therapy and / or imaging, diagnosis and / or immunostaining.

[0004] Background of the invention

[0005] An interesting tumor associated antigen which could be used as target in antibody therapy is the PTK7 antigen. PTK7 is one of several receptor tyrosine kinases (RTKs), although PTK7 is an atypical RTK without intrinsic tyrosine kinase activity, PTK7 -dependent signaling plays key roles during embryogenesis and carcinogenesis.

[0006] Tyrosine-protein kinase-like 7 also known as colon carcinoma kinase 4 (CCK4) is a receptor tyrosine kinase that in humans is encoded by the PTK7 gene.

[0007] PTK7 is an evolutionarily conserved transmembrane receptor with a broad range of functions in tissue development and homeostasis (Berger et al., 2017). Originally identified as a gene upregulated in colon carcinoma cells and accordingly named colon carcinoma kinase 4 (CCK-4) (Mossie et al., 1995) it has later been shown to affect various aspects of cell-cell communication and movement. PTK7 controls tissue morphogenesis and patterning by affecting cell polarity, migration as well as tissue regeneration and wound healing (Lu et al., 2004; Shnitsar and Borchers, 2008; Caddy et al., 2010; Lee et al., 2011 ; Lander and Petersen, 2016). Additionally, PTK7 function in adult tissue homeostasis is demonstrated by the fact that misregulation of PTK7 expression correlates with development of cancer and its progression to metastasis in various cellular contexts (Dunn and Tolwinski, 2016). Thus, PTK7 could be a target for cancer therapy. The expression of PTK7, which is upregulated in various malignancies, is negatively correlated with disease-free survival and / or overall survival in patients with cancer (Won Oh et al., 2022). PTK7 potentiates oncogenic signaling by functioning as a co-receptor for active RPTKs, such as FGFR1 (Shin et al., 2019).

[0008] The use of monoclonal antibodies has increased steadily since its discovery in the 1970s and today it is a multi-billion industry. Antibody based treatments have been developed against a number of cancer associated antigens including Her-2 and CD20 and today monoclonal antibodies constitutes an important class of therapeutics.

[0009] The mechanisms of action of monoclonal antibodies in targeted therapy are diverse. Some therapeutic antibodies act by arresting proliferation of target cell by binding to the target antigen receptor on the cell surface. Other monoclonal antibodies have been developed (chimeric, humanized, fully human) to interact with human immune effectors (e.g. complement factors, Natural Killer cells) to stimulate targeted tumor cell kill by these immune effectors. Monoclonal antibodies can be further developed as targeting entities by conjugation to nanoparticles or microparticles made from polymers or proteins or inorganic crystals or a combination thereof, carrying cytotoxic drugs or radioactive molecules. The monoclonal antibodies can also be directly conjugated to drugs (Antibody Drug Conjugates) (Sinha et al., 2006, Peer et al., 2007, Sievers & Senter, 2013)

[0010] Monoclonal antibodies have been developed against the PTK7 antigen in the past and in vitro and in vivo testing of targeting with anti-PTK7 monoclonal antibodies has shown a considerable promise.

[0011] Receptor tyrosine kinases are frequently overexpressed or aberrantly activated in human cancers, and preferable targets for antibody cancer therapy. This also applies when antibodies are used with payloads such as cytostatic or radionuclides, owing to accessibility of and internalization of the antibody with the therapeutic payload. For instance, the HER2-targeting trastuzumab emtansine is approved for the treatment of breast cancer patients, whereas other RTKs, such as AXL, EGFR, EPHA2, ERBB3, FGFR2, FGFR3, FLT3, KIT, MET and also PTK7, are under development for the treatment of cancer patients. There is only one anti-PTK7 antibody in clinical development for cancer treatment, and this is the ADC cofetuzumab pelidotin. No PTK7 targeting clinical stage radionuclide therapeutics have been identified so far.

[0012] Thus, there is a need for further development of PTK7 targeting clinical stage therapeutics, imaging etc.

[0013] SUMMARY

[0014] In its broadest aspect, the present invention relates to an antibody or a fragment of an antibody, wherein the antibody is comprising a heavy chain and a light chain, wherein the heavy chain comprises a CDRH1 having an amino acid sequence that is 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 10, a CDRH2 having an amino acid sequence that is 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 11 , and a CDRH3 having an amino acid sequence that is 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 12, and the light chain comprises a CDRL1 having an amino acid sequence that is 90%, such as for example 93, 95, 98, or 99% identical to any one of SEQ ID NO: 16, a CDRL2 having an amino acid sequence that is 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 17, and a CDRL3 having an amino acid sequence that is 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 18. One or more embodiments of the invention relates to an antibody or fragment of an antibody according to the invention, wherein the heavy chain comprises a CDRH1 having an amino acid sequence according to any one of SEQ ID NO: 10, a CDRH2 having an amino acid sequence according to any one of SEQ ID NO: 11 , and a CDRH3 having an amino acid sequence according to any one of SEQ ID NO: 12, and the light chain comprises a CDRL1 having an amino acid sequence according to any one of SEQ ID NO: 16, a CDRL2 having an amino acid sequence according to any one of SEQ ID NO: 17, and a CDRL3 having an amino acid sequence according to any one of SEQ ID NO: 18.

[0015] One or more embodiments of the invention relates to an antibody or fragment of an antibody according to the invention, wherein the heavy chain comprises an amino acid sequence according to any one of SEQ ID NO: 3, and the light chain comprises an amino acid sequence according to SEQ ID NO: 4.

[0016] One or more embodiments of the invention relates to an antibody or fragment of an antibody according to the invention, wherein the heavy chain has an amino acid sequence according to SEQ ID NO: 3, and the light chain has an amino acid sequence according to SEQ ID NO: 4.

[0017] One or more aspects of the invention relates to an antibody fragment of an antibody according to the invention, wherein the antibody fragment is an Fv, scFv, Fab, F(ab')2, Fab', or scFv-Fc antibody fragment.

[0018] One or more aspects of the invention relates to an antibody-drug conjugate comprising an antibody according to or an antibody fragment according to the invention.

[0019] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention and further comprising a linker and / or a chelator.

[0020] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the antibody or antibody fragment is conjugated to a chelator and / or a linker.

[0021] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the chelator and / or a linker binds a drug or radionuclide.

[0022] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the chelator and / or a linker forms a complex with a drug or radionuclide.

[0023] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the chelator is selected as one from the group consisting of DOTA derived chelators such as: p-SCN-bn-DOTA, DOTA-NHS-ester, p-SCN-Bn- DOTAM (TCMC); bifunctional linear chelators like p-SCN-Bn-DTPA and CHX-A"-DTPA, and further Me-3,2-HOPO or other 3- hydroxy-N-methyl-2-pyridinone derived chelates, Crypt-OH or other derivatives of [2.2.2]-cryptands and H2macropa-NCS or other macropa derived chelates. One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the chelator is p-SCN-Bn-TCMC.

[0024] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the chelator is p-SCN-bn-DOTA.

[0025] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the antibody-drug conjugate comprises a radionuclide.

[0026] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the radionuclide is selected as any one from the group consisting of211At,213Bi,212Bi,212Pb,225Ac,227Th,223Ra,224Ra,90Y,161Tb,186Re,188Re,199Au,194lr,166Ho,159Gd,153Sm,149Pm,142Pr,111Ag,109Pd,77As,67Cu,64Cu,47Sc, and177Lu.

[0027] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention comprising a radionuclide, wherein the radionuclide is212Pb.

[0028] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention comprising a radionuclide, wherein the radionuclide is177Lu.

[0029] One or more aspects of the invention relates to a nucleic acid sequence encoding the antibody or the antibody fragment according to the invention.

[0030] One or more embodiments of the invention relates to the nucleic acid construct comprising the nucleic acid sequence according to the invention.

[0031] One or more embodiments of the invention relates to a host cell comprising the nucleic acid sequence or the nucleic acid construct according to the invention.

[0032] One or more aspects of the invention relates to a method for producing an antibody or an antibody fragment according to the invention, comprising providing a host cell comprising the nucleic acid sequence or the nucleic acid construct according to the invention, culturing the host under conditions ensuring expression of the nucleic acid or nucleic acid construct, and recovering the antibody or antibody fragment.

[0033] One or more aspects of the invention relates to a method for producing a chelator-antibody conjugate labelled with a radionuclide according to the invention comprising the steps of obtaining an antibody or antibody fragment according to the invention, mixing said antibody or antibody fragment with a chelator to obtain a chelator antibody conjugate, admixing said chelator-antibody conjugate or chelator antibody fragment conjugate with a radionuclide, and recovering a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate. One or more aspects of the invention relates to a composition comprising an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention.

[0034] One or more aspects of the invention relates to a pharmaceutical composition comprising an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelatorantibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention.

[0035] One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use as a medicament.

[0036] One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of cancer.

[0037] One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of a PTK7 expressing cancer selected from the group consisting of but not limited to ovarian cancer, cervical cancer, endometrial cancer, breast cancer including triple-negative breast cancer, renal cancer, bladder cancer including urothelial cancer, colorectal cancer, mesothelioma, osteosarcoma, brain cancer, including glioma, head and neck cancer, liver cancer including hepatocellular carcinoma, lung cancer including non-small-cell lung cancer, lymphoma, myeloma, leukemia, melanoma, neuroendocrine cancer including carcinoid, pancreatic cancer, prostate cancer, testis cancer, skin cancer, gastric cancer, squamous cell carcinoma including esophageal squamous cell carcinoma, and thyroid cancer..

[0038] One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of ovarian cancer.

[0039] One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the diagnosis, monitoring or imaging of cancer.

[0040] One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use as an imaging agent.

[0041] One or more aspects of the invention relates to kit for producing a radionuclide labelled chelatorantibody conjugate or radionuclide labelled chelator-antibody fragment conjugate, the kit comprising an antibody or an antibody fragment according to the invention, a solution of a radionuclide selected as any one of the group consisting of211At,213Bi,212Bi,212Pb,225Ac,227Th, "Y,161Tb,186Re,188Re,199Au,194lr,166Ho,159Gd,153Sm,149Pm,142Pr,111Ag,109Pd,77As,67Cu,64Cu,47Sc, and177Lu, and optionally instructions for producing the radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate using the materials provided in the kit.

[0042] One or more aspects of the invention relates to kit for producing a radionuclide labelled chelatorantibody conjugate or radionuclide labelled chelator-antibody fragment conjugate, the kit comprising a chelator-antibody conjugate or complex, wherein the antibody is an antibody or an antibody fragment according to the invention, a solution of a radionuclide selected as any one of the group consisting of211At,213Bi,212Bi,212Pb,225Ac,227Th, "Y,161Tb,186Re,188Re,199Au,194lr,166Ho,159Gd,153Sm,149Pm,142Pr,111Ag,109Pd,77As,67Cu,64Cu,47Sc, and177Lu, and optionally instructions for producing the radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate using the materials provided in the kit.

[0043] One or more embodiments of the invention relates to a kit according to the invention, wherein the chelator is selected as any one of p-SCN-bn-DOTA and p-SCN-Bn-TCMC and the radionuclide is selected as any one of177Lu or212Pb.

[0044] One or more embodiments of the invention relates to a kit according to the invention, wherein the chelator is selected as TCMC and the radionuclide is selected as177Lu.

[0045] One or more embodiments of the invention relates to a kit according to the invention, wherein the chelator is selected as Bn-DOTA and the radionuclide is selected as212Pb.

[0046] DETAILED DESCRIPTION

[0047] The present invention is based on the pre-clinical data disclosed in the examples, which shows that an anti-PTK7 antibody, such as 01-1 , with binding affinity, in vivo targeting potential, data supporting internalization as well as in vivo anti-tumor effect with a therapeutic radionuclide, makes an anti-PTK7 antibody, such as 01-1 , an interesting targeting agent for development of a fullantibody or antibody fragment for delivery of a radionuclide for site-direct cancer therapy.

[0048] Thus, the present invention relates to an antibody or a fragment of an antibody or antibody derivative thereof, wherein the antibody is comprising a heavy chain and a light chain, wherein the heavy chain comprises a CDRH1 having an amino acid sequence that is 80%, such as 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 10, a CDRH2 having an amino acid sequence that is 80%, such as 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 11 , and a CDRH3 having an amino acid sequence that is 80%, such as 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 12, and the light chain comprises a CDRL1 having an amino acid sequence that is 80%, such as 90%, such as for example 93, 95, 98, or 99% identical to any one of SEQ ID NO: 16, a CDRL2 having an amino acid sequence that is 80%, such as 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 17, and a CDRL3 having an amino acid sequence that is 80%, such as 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 18.

[0049] The term “sequence identity of [a certain] %” in the context of two or more nucleic acid or amino acid sequences means that the two or more sequences have nucleic acids or amino acid residues in common in the given percent, when compared and aligned for maximum correspondence over a comparison window or designated sequences of nucleic acids or amino acids (e.g., the sequences have at least 90 percent (%) identity). Percent identity of nucleic acid or amino acid sequences can be measured using a BLAST 2.0 sequence comparison algorithm with default parameters, or by manual alignment and visual inspection (see e.g. http: / / www.ncbi.nlm.nih.gov / BLAST / ). This definition also applies to the complement of a test sequence and to sequences that have deletions and / or additions, as well as those that have substitutions. An example of an algorithm that is suitable for determining percent identity, sequence similarity and for alignment is the BLAST 2.2.20+ algorithm, which is described in Altschul et al. Nucl. Acids Res. 25, 3389 (1997). BLAST 2.2.20+ is used to determine percent sequence identity for the nucleic acids and proteins of the disclosure. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (http: / / www.ncbi.nlm.nih.gov / ). Examples of commonly used sequence alignment algorithms are

[0050] CLUSTAL Omega (http: / / www.ebi.ac.uk / Tools / msa / clustalo / ),

[0051] EMBOSS Needle (http: / / www.ebi.ac.uk / Tools / psa / emboss_needle / ),

[0052] MAFFT (http: / / mafft.cbrc.jp / alignment / server / ), or

[0053] MUSCLE (http: / / www.ebi.ac.uk / Tools / msa / muscle / ).

[0054] In that regard, in one or more embodiment(s) the sequence identity of a sequence is at least 80 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 81 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 82 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 83 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 84 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 85 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 86 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 87 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 88 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 89 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 90 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 91 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 92 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 93 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 94 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 95 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 96 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 97 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 98 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 99 % identical compared to a reference sequence. In one or more embodiment(s) the sequence identity of a sequence is at least 99,9 % identical compared to a reference sequence.

[0055] In one or more embodiment(s) the sequence identity of a sequence is 100 % identical to a reference sequence.

[0056] One or more embodiments of the invention relates to an antibody or fragment of an antibody according to the invention, wherein the heavy chain comprises a CDRH1 having an amino acid sequence according to any one of SEQ ID NO: 10, a CDRH2 having an amino acid sequence according to any one of SEQ ID NO: 11 , and a CDRH3 having an amino acid sequence according to any one of SEQ ID NO: 12, and the light chain comprises a CDRL1 having an amino acid sequence according to any one of SEQ ID NO: 16, a CDRL2 having an amino acid sequence according to any one of SEQ ID NO: 17, and a CDRL3 having an amino acid sequence according to any one of SEQ ID NO: 18.

[0057] One or more embodiments of the invention relates to an antibody or fragment of an antibody according to the invention, wherein the heavy chain comprises an amino acid sequence according to any one of SEQ ID NO: 3, and the light chain comprises an amino acid sequence according to SEQ ID NO: 4.

[0058] One or more embodiments of the invention relates to an antibody or fragment of an antibody according to the invention, wherein the heavy chain has an amino acid sequence according to SEQ ID NO: 3, and the light chain has an amino acid sequence according to SEQ ID NO: 4.

[0059] A typical antibody is composed of two immunoglobulin (Ig) heavy chains and two Ig light chains.

[0060] Several different types of heavy chain exist that define the class or isotype of an antibody. These heavy chain types vary between different animals.

[0061] The immunoglobulin light chain is the small polypeptide subunit of an antibody (immunoglobulin).

[0062] There are two types of light chain in humans (as in other mammals), kappa (K) chain, encoded by the immunoglobulin kappa locus on chromosome 2 and the lambda (A) chain, encoded by the immunoglobulin lambda locus on chromosome 22.

[0063] Antibodies are produced by B lymphocytes, each expressing only one class of light chain.

[0064] Once set, light chain class remains fixed for the life of the B lymphocyte.

[0065] In a healthy individual, the total kappa to lambda ratio is roughly 2:1 in serum (measuring intact whole antibodies) or 1 :1.5 if measuring free light chains, with a highly divergent ratio indicative of neoplasm.

[0066] The exact normal ratio of kappa to lambda ranges from 0.26 to 1.65.

[0067] Both the kappa and the lambda chains can increase proportionately, maintaining a normal ratio.

[0068] An antibody, antibody fragment or antibody derivative thereof, may comprise a light chain variable domain (VL) and / or a heavy chain variable domain (VH). In some cases, an antibody, antibody fragment or antibody derivative thereof comprises a light chain variable domain (VL) and no heavy chain variable domain (VH). In other cases, an antibody, antibody fragment or antibody derivative thereof comprises a heavy chain variable domain (VH) and no light chain variable domain (VL)

[0069] Thus, in one or more embodiment(s), the antibody, antibody fragment or antibody derivative thereof, comprises a light chain variable domain (VL) and / or a heavy chain variable domain (VH).

[0070] In one embodiment, the antibody, antibody fragment or antibody derivative thereof comprises a lambda or kappa light chain constant domain.

[0071] In one or more embodiment(s) the light chain constant domain is of human origin.

[0072] In one embodiment, the antibody, antibody fragment or antibody derivative thereof is comprises a kappa light chain constant domain encoded by a nucleic acid sequence of SEQ ID NO: 5, or a functional homologue thereof having an amino acid sequence which is at least 80 % identical, such as at least 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 95 %, 96 %, 97 %, 98 %, 99 % or 99.9 % identical to any of the variants of SEQ ID NO: 5.

[0073] Similarly, to the light chain constant domain, the heavy chain domain can be optimised by grafting of the CDR regions onto e.g. human heavy chain acceptor sequences. Thus, in one or more embodiment(s) of the present disclosure the antibody, antibody fragment or antibody derivative thereof is defined by i) a constant heavy chain is selected from the group consisting of IgG 1 , lgG2, lgG3 and lgG4 chain, and ii) a constant light chain is a kappa or a lambda chain, wherein heavy chain variable regions according to the present disclosure are grafted onto the constant heavy chain, and light chain variable regions according to the present disclosure are grafted onto the constant light chain.

[0074] In one embodiment, the antibody, antibody fragment or antibody derivative thereof is comprises a kappa heavy chain constant domain encoded by a nucleic acid sequence of SEQ ID NO: 6, or a functional homologue thereof having an amino acid sequence which is at least 80 % identical, such as at least 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 95 %, 96 %, 97 %, 98 %, 99 % or 99.9 % identical to any of the variants of SEQ ID NO:

[0075] 6.

[0076] In a further embodiment, the antibody, antibody fragment or antibody derivative thereof is an antibody that comprises an IgG 1 , lgG2, lgG3 or lgG4, IgM, IgA, IgE and / or IgD heavy chain constant domain of human origin.

[0077] In another embodiment, the antibody, antibody fragment or antibody derivative thereof comprises a lambda and / or kappa light chain constant domain of human origin and / or an IgG 1 , lgG2, lgG3 or lgG4, IgM, IgA, IgE or IgD heavy chain constant domain of human origin.

[0078] In another embodiment, the antibody, antibody fragment or antibody derivative thereof comprises a lambda or kappa light chain constant domain of human origin and / or an IgG 1 , lgG2, lgG3 or lgG4, IgM, IgA, IgE or IgD heavy chain constant domain of human origin.

[0079] In another embodiment, the antibody, antibody fragment or antibody derivative thereof is an antibody that comprises a lambda or kappa light chain constant domain of human origin and an IgG 1 , lgG2, lgG3 or lgG4, IgM, IgA, IgE or IgD heavy chain constant domain of human origin.

[0080] In a further embodiment, the antibody, antibody fragment or antibody derivative thereof is an antibody that comprises a kappa light chain constant domain of human origin and an IgG 1 , lgG2, lgG3 or lgG4 heavy chain constant domain of human origin.

[0081] In a further embodiment, the antibody, antibody fragment or antibody derivative thereof is an antibody that comprises a kappa light chain constant domain of human origin and an IgG 1 or lgG3 heavy chain constant domain of human origin.

[0082] In a preferred embodiment, the antibody, antibody fragment or antibody derivative thereof is an antibody that comprises a kappa light chain constant domain of human origin and an IgG 1 heavy chain constant domain of human origin.

[0083] The examples (9A, 9B, and 10) show that e.g. chimeric OI-1 with IgG 1 has a positive profile. Immunotherapy using monoclonal antibodies (mAbs) has been emerging as a safe and selective method for the treatment of cancer and other diseases.

[0084] In particular, the role of monoclonal antibodies in therapies that are based on B-cell depletion, e.g. in the treatment of B-cell malignancies, has expanded since the introduction of rituximab, an antibody that is directed against the CD20 antigen on the B-cell surface.

[0085] Thus, in one or more preferred exemplified embodiment(s), the antibody, antibody fragment or antibody derivative thereof is a monoclonal antibody.

[0086] A traditional antibody comprises two disulphide bridge linked heavy chains and two light chains, linked to the heavy chain via disulphide bridges. The heavy chain, in general comprise a variable domain (VH) and potentially three constant domains CH1-CH3, wherein CH1 and CH2 is linked via a hinge region comprising one or more cysteines partially responsible for heavy chain dimerization. CH2 and CH3 also comprises cysteines also partially responsible for heavy chain dimerization. The light chain comprises a variable domain (VL) and a constant domain (CL). The heavy chain CH1 and light chain CL are linked via one or more cysteine residues forming one or more disulphide bridge(s).

[0087] Due to the many different domains and chain linkage options several different antibody fragment variants have emerged.

[0088] Modifying antibody features such as molecular size, valency, binding affinity, and pharmacokinetics allows for the development of antibody fragments with tailor-made properties for a variety of clinical applications. Variation in molecular size and binding properties among antibody fragments and antibody derivates thereof is considered to possess a central role in the tissue distribution of targeting molecules.

[0089] One or more aspects of the invention relates to an antibody fragment of an antibody according to the invention, wherein the antibody fragment is an Fv, scFv, Fab, F(ab')2, Fab', or scFv-Fc antibody fragment.

[0090] The antigen-binding fragment (Fab), according to the present disclosure, is a region on an antibody that binds to antigens. It is composed of one constant and one variable domain of each of the heavy and the light chain, thus a Fab fragment does not contain an Fc fragment. The variable domains comprise the antigen-binding site, comprising the CDRs.

[0091] Fab fragments may be prepared from an IgG like antibody by enzymatic degradation targeting the hinge region of said antibody. Alternatively, a Fab fragment is produced in a host cell, comprising a nucleotide encoding the Fab fragment, thus, only producing the portion of the antibody fragment that is the Fab fragment. Thus, in one or more embodiments, the antibody, antibody fragment or antibody derivative thereof is a Fab fragment, comprising a heavy chain variable domain (VH) that comprises any one of the amino acid sequence variants of SEQ ID NO: 3 disclosed herein, and the light chain variable domain (VL) comprises any one of the amino acid sequence variants of SEQ ID NO: 4, or functional homologues thereof having an amino acid sequence which is at least 80 % identical, such as at least 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 95 %, 96 %, 97 %, 98 %, 99 % or 99.9 % identical to any of the variants of SEQ ID NO: 1 and / or SEQ ID NO: 4.

[0092] In one or more further embodiments, the antibody, antibody fragment or antibody derivative thereof is a Fab fragment, comprising a heavy chain variable domain (VH) that comprises the amino acid sequence of SEQ ID NO: 3 and the light chain variable domain (VL) comprises the amino acid sequence of SEQ ID NO: 4.

[0093] A Fab’ fragment, according to the present disclosure, is a Fab fragment, further comprising at least a portion of the hinge region of a traditional antibody, but which does not comprise a disulphide bridge responsible for dimerization of the individual fragments.

[0094] Fab’ fragments may be prepared from an IgG like antibody by enzymatic degradation targeting the hinge region of said antibody. Alternatively, a Fab’ fragment is produced in a host cell, comprising a nucleotide encoding the Fab’ fragment, thus, only producing the portion of the antibody fragment that is the Fab’ fragment, wherein the Fab’ fragment is postprocessed in order to reduce disulphide bridges formed between individual Fab’ fragments.

[0095] Thus in one or more embodiments, the antibody, antibody fragment or antibody derivative thereof is a Fab’ fragment, comprising a heavy chain variable domain (VH) that comprises any one of the amino acid sequence variants of SEQ ID NO: 3 and the light chain variable domain (VL) comprises any one of the amino acid sequence variants of SEQ ID NO: 4, or functional homologues thereof having an amino acid sequence which is at least 80 % identical, such as at least 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 95 %, 96 %, 97 %, 98 %, 99 % or 99.9 % identical to any of the variants of SEQ ID NO: 3 and / or SEQ ID NO: 4.

[0096] Thus, in one or more further embodiments, the antibody, antibody fragment or antibody derivative thereof is a Fab’ fragment, comprising a heavy chain variable domain (VH) that comprises the amino acid sequence of SEQ ID NO: 3 and the light chain variable domain (VL) comprises the amino acid sequence of SEQ ID NO: 4.

[0097] A Fab’ fragment, according to the present disclosure, is a Fab fragment, further comprising at least a portion of the hinge region of a traditional antibody i.e., a Fab’ fragment, wherein, the disulphide bridges linking individual Fab’ fragments is not reduced, thus making a dimeric antibody Fab’ fragment, denoted as F(ab’)2. Thus, in one or more embodiment(s), the disulphides of the antibody fragment hinge region is reduced, resulting in a Fab’ antibody fragment. In one or more embodiment(s) the disulphides are oxidized resulting in a F(ab’)2 antibody fragment.

[0098] In that regard, in one or more embodiments, the antibody, antibody fragment or antibody derivative thereof is a F(ab’)2 fragment, comprising one or more heavy chain variable domain(s) (VH) that comprises any one or more of the amino acid sequence variants of SEQ ID NO: 3 and one or more light chain variable domain(s) (VL) comprises any one or more of the amino acid sequence variants of SEQ ID NO: 4, or functional homologues thereof having an amino acid sequence which is at least 80 % identical, such as at least 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 95 %, 96 %, 97 %, 98 %, 99 % or 99.9 % identical to any of the variants of SEQ ID NO: 3 and / or SEQ ID NO: 4.

[0099] Thus, in one or more further embodiments, the antibody, antibody fragment or antibody derivative thereof is a F(ab’)2 fragment, comprising a heavy chain variable domain (VH) that comprises the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain (VL) comprises the amino acid sequence of SEQ ID NO: 4.

[0100] Alternatively, the Fab or Fab’ fragments are chemically linked, by a chemical linker, such as but not limited to a disuccinimidyl suberate (DSS) linker, N-Hydroxysuccinimide-Polyethyleneglycol (NHS- PEG) linker or similar chemical linker, resulting in a chemically linked F(ab)2 or F(ab’)2 fragment.

[0101] In that regard the F(ab)2 or F(ab’)2 may be a monospecific or bi-specific antibody fragment.

[0102] Fab, Fab’, F(ab)2 or F(ab’)2 may also be combined into a F(ab)3 fragment comprising three individual Fab or Fab’, or a F(ab)2, a F(ab’)2 and a Fab or Fab’ fragment, thus, comprising three light chains and three heavy chain fragments, linked into a tripart fragment.

[0103] The F(ab)s fragment may be assembled by disulphide linkage or chemical linkage as disclosed herein.

[0104] In that regard, in one or more embodiments, the antibody, antibody fragment or antibody derivative thereof is a F(ab)s fragment, comprising one or more heavy chain variable domain (VH) that comprises any one or more of the amino acid sequence variants of SEQ ID NO: 3 [and one or more light chain variable domain (VL) comprises any one or more of the amino acid sequence variants of SEQ ID NO: 4, or functional homologues thereof having an amino acid sequence which is at least 80 % identical, such as at least 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 95 %, 96 %, 97 %, 98 %, 99 % or 99.9 % identical to any of the variants of SEQ ID NO: 3 and / or SEQ ID NO: 4. Thus, in one or more further embodiments, the antibody, antibody fragment or antibody derivative thereof is a F(ab)s fragment, comprising a heavy chain variable domain (VH) that comprises the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain (VL) comprises the amino acid sequence of SEQ ID NO: 4.

[0105] In that regard the F(ab)3 may be a monospecific, bi-specific or trispecific antibody fragment.

[0106] An scFV fragment, according to the present disclosure, is an antibody fragment, comprising a heavy chain fragment comprising a variable domain (VH) and optionally a constant domain (CH) and a light chain fragment comprising a variable domain (VL) and optionally a constant light chain domain (CL), wherein the light chain and the heavy chain fragments are linked by a linker, thus making a single fragment. Such as linker may be a chemical linker as described in the present disclosure, an amino acid linker, such as but not limited to a poly-Gly-Ser linker or it may be a combination of a chemical and amino acid linker.

[0107] In that regard, in one or more embodiments, the antibody, antibody fragment or antibody derivative thereof is a scFV fragment, comprising a heavy chain variable domain (VH) that comprises any one of the amino acid sequence variants of SEQ ID NO: 3 and the light chain variable domain (VL) which comprises any one of the amino acid sequence variants of SEQ ID NO: 4], or functional homologues thereof having an amino acid sequence which is at least 80 % identical, such as at least 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 95 %, 96 %, 97 %, 98 %, 99 % or 99.9 % identical to any of the variants of SEQ ID NO: 3 and / or SEQ ID NO: 4, wherein the fragment further comprises a linker linking the heavy chain and the light chain.

[0108] Thus, in one or more further embodiments, the antibody, antibody fragment or antibody derivative thereof is an scFV fragment, comprising a heavy chain variable domain (VH) that comprises the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain (VL) comprises the amino acid sequence of SEQ ID NO: 4.

[0109] Individual scFV fragment may also be linked to form a dimeric, trimeric or tetrameric scFV construct i.e., SCFV2, scFVs, scFV4. Such a link may also be a disulphide link. SCFV2, scFVs, scFV4 are also referred to as diabodies, tribadies or tetrabodies.

[0110] In yet another embodiment, the antibody, antibody fragment or antibody derivative thereof the antibody fragment is a diabody, triabody, or tetrabody.

[0111] An scFV fragment, according to the present disclosure, is an antibody fragment, comprising a heavy chain fragment comprising a variable domain (VH) and more than one constant domain (CH), such as but not limited to CHi and CH2 or CH1, CH2 and CH3, and a light chain fragment comprising a variable domain (VL) and optionally a constant light chain domain (CL), wherein the light chain and the heavy chain fragments are linked by a linker, thus making a single fragment. Such as linker may be a chemical linker as described in the present disclosure, an amino acid linker, such as but not limited to a poly-Gly-Ser linker or it may be a combination of a chemical and amino acid linker.

[0112] In that regard, in one or more embodiments, the antibody, antibody fragment or antibody derivative thereof is a scFV-Fc fragment, comprising a heavy chain variable domain (VH) that comprises any one of the amino acid sequence variants of SEQ ID NO: 3 and the light chain variable domain (VL) which comprises any one of the amino acid sequence variants of SEQ ID NO: 4], or functional homologues thereof having an amino acid sequence which is at least 80 % identical, such as at least 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 95 %, 96 %, 97 %, 98 %, 99 % or 99.9 % identical to any of the variants of SEQ ID NO: 3 and / or SEQ ID NO: 4, wherein the fragment further comprises a linker linking the heavy chain and the light chain.

[0113] Thus, in one or more further embodiments, the antibody, antibody fragment or antibody derivative thereof is an scFV-Fc fragment, comprising a heavy chain variable domain (VH) that comprises the amino acid sequence of SEQ ID NO: 3 and a light chain variable domain (VL) comprises the amino acid sequence of SEQ ID NO: 4.

[0114] A scFV-Fc fragment comprising a CHi and CH2 domain may also be referred to as a minibody.

[0115] Thus, in one or more embodiment(s), the antibody, antibody fragment or antibody derivative thereof the antibody fragment is a minibody.

[0116] The present disclosure relates to an antibody, antibody fragment or antibody derivative thereof, wherein the antibody, antibody fragment or antibody derivative thereof is optimized for binding to PTK7. Binding of a PTK7-specific antibody, antibody fragment or antibody derivative thereof, to cancer cells may trigger various mechanisms of action:

[0117] - After the antibody, antibody fragment or antibody derivative thereof, binds to the extracellular domain of the PTK7 antigen, it may activate the complement cascade and lyse the targeted cell.

[0118] - An anti-PTK7 antibody, antibody fragment or antibody derivative thereof, may mediate antibodydependent cell-mediated cytotoxicity (ADCC), antibody dependent cellular phagocytosis (ADCP) and / or complement-dependent cytotoxicity (CDC), to the target cell, which occurs after the Fc portion of the bound antibody is recognized by appropriate receptors on cytotoxic cells of the immune system.

[0119] - The antibody may alter the ability of B-cells to respond to antigen or other stimuli.

[0120] Finally, anti-PTK7 antibody, antibody fragment or antibody derivative thereof, may initiate programmed cell death (apoptosis). Thus, in one embodiment, the antibody, antibody fragment or antibody derivative thereof is a PTK7 targeting molecule.

[0121] In one embodiment, the antibody, antibody fragment or antibody derivative thereof is an anti-PTK7 antibody, antibody fragment or antibody derivative thereof according to the present disclosure.

[0122] In another embodiment, the antibody, antibody fragment or antibody derivative thereof is a polyclonal anti-PTK7 antibody.

[0123] In one or more exemplified embodiment(s), the antibody, antibody fragment or antibody derivative thereof is a monoclonal anti-PTK7 antibody.

[0124] In one or more embodiment(s) the antibody, antibody fragment or antibody derivative thereof, forms IgG complexes in the presence of PTK7.

[0125] In the present disclosure an antibody derivate relates to antibody derivatives that make use of selected parts of an antibody resulting in molecules with novel biological activity and rationally designed mechanisms of action.

[0126] In that regard, in one or more embodiment(s) of the present disclosure relates to an antibody derivate.

[0127] In one embodiment, the antibody, antibody fragment or antibody derivative thereof is a chimeric, human or humanized antibody.

[0128] Antibody glycosylation defines the functional potential of the antibody by delineating the structure of the antibody Fc region and determining which Fc receptors it can bind to in order to recruit effector cells.

[0129] The effector functions that antibodies mediate, including cytotoxicity and phagocytosis, are critical for protection against and prevention of many diseases.

[0130] Antibody glycosylation has been harnessed to improve the efficacy of monoclonal therapeutics.

[0131] Antibody glycosylation can be modulated by vaccination, indicating that rational immunogen design could seek to elicit a specific antibody glycosylation response.

[0132] Thus, in one or more exemplified embodiments, the antibody, antibody fragment or antibody derivative thereof is glycosylated.

[0133] Glycoengineered therapeutic antibodies lacking core fucose residue from the Fc N-glycans exhibit strong ADCC at lower concentrations with much higher efficacy compared to fucosylated counterparts and can evade the inhibitory effect of serum immunoglobulin G (IgG) on ADCC through its high binding to gamma receptor Illa (Fc FcyRllla). Thus, in an embodiment, said glycosylation of said antibody, antibody fragment or antibody derivative thereof is fucose deficient.

[0134] In one or more embodiment(s) said fucose deficient antibody, antibody fragment or antibody derivative thereof have an enhanced and / or improved induction of antibody-dependent cell- mediated cytotoxicity (ADCC), compared to a non-fucose deficient antibody, antibody fragment or antibody derivative thereof.

[0135] In that regard an enhanced and / or improved induction of antibody-dependent cell-mediated cytotoxicity (ADCC), may relate an at least 1.01-fold increase, such as at least 1.1 , 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 2, or 5-fold increase, in induction of antibody-dependent cell-mediated cytotoxicity (ADCC), compared to a non-fucose deficient antibody, antibody fragment or antibody derivative thereof.

[0136] In another exemplified embodiment said fucose deficient antibody, antibody fragment or antibody derivative thereof have an enhanced and / or improved induction of antibody-dependent cell- mediated cytotoxicity (ADCC), compared to a non-fucose deficient antibody, antibody fragment or antibody derivative thereof.

[0137] Said fucose deficiency may be obtained in several ways, such as but not limited to introduction of a GDP-4-keto-6-deoxy mannose reductase in the production host cell. This protein is a bacterial GDP-4-keto-6-deoxy mannose reductase (RMD) that depletes the cytosolic pool of GDP-4-keto-6- deoxy mannose, which is a precursor for the synthesis of fucose. This precursor is being transformed to GDP-D-Rhamnose - an important for bacteria, but inactive sugar in mammalian cell. An alternative may also be the introduction of 4-b-N-acetylglucosaminyltransferase (GnT-lll) and Golgi a-mannosidase II (aManll), which also inhibits fucosylation, thus producing a fucose deficient product.

[0138] Thus, in one or more embodiment(s), the production host cell is engineered such that it expresses and / or overexpresses a GDP-4-keto-6-deoxy mannose reductase and / or a 4-b-N- acetylglucosaminyltransferase (GnT-lll) and / or a Golgi a-mannosidase II (aManll).

[0139] In the present disclosure, the antibody, antibody fragment or antibody derivative thereof targeting human PTK7 are high affinity molecules. In general, high affinity molecules are highly preferable in the development of novel therapeutics, as a high affinity may limit off-target side effects, may enhance the on-target effects and may reduce the dosage needed in order to obtain the desired effect.

[0140] Thus, in one or more embodiment(s), the antibody, antibody fragment or antibody derivative thereof has an affinity for human PTK7 expressing cells below 10 nM, such as below 9 nM, 8 nM, 7 nM, 6 nM, 5 nM, 4 nM, 3 nM, 2 nM, or 1 nM and / or such as below 1 nM, such as below 900 pM, 800 pM, 700 pM, 600 pM, 500 pM, 400 pM or 300 pM.

[0141] In one or more preferred embodiment(s), the antibody, antibody fragment or antibody derivative thereof comprises the heavy chain variable domain (VH) comprises the amino acid sequence of SEQ ID NO: 3 and the light chain variable domain (VL) comprises the amino acid sequence variants of SEQ ID NO: 4 and wherein the antibody, antibody fragment or antibody derivative thereof have an affinity for human PTK7 expressing cells below 2 nM such as such as below 1 nM, such as below 900 pM, 800 pM, 700 pM, or 600 pM.

[0142] The antibodies and antibody fragment of an antibody according to the invention show promising effects on PTK7 as a target with binding affinity, in vivo targeting potential, data supporting internalization as well as in vivo anti-tumor effect with a therapeutic radionuclide,

[0143] These effects are shown in the experiments of the present disclosure.

[0144] One or more aspects of the invention relates to a nucleic acid sequence encoding the antibody or the antibody fragment according to the invention.

[0145] In one or more embodiments(s) of the present disclosure a nucleic acid sequence encodes one or more amino acid sequences according to the present disclosure.

[0146] In one embodiment, a nucleic acid sequence encoding the light chain of the antibody, antibody fragment or antibody derivative thereof according to the present disclosure, encodes an amino acid sequence according to any one of SEQ ID NOs: 2, or a functional homologue thereof having amino acid sequence which is at least 70 % identical, such as at least 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 95 %, 96 %, 97 %, 98 %, 99 % or 99.9 % identical to any of the variants of SEQ ID NO: 2.

[0147] In a embodiment, the nucleic acid sequence encoding the light chain of the antibody, antibody fragment or antibody derivative thereof according to the present disclosure is a nucleic acid sequence of SEQ ID NO: 2.

[0148] In one embodiment, the nucleic acid sequence encoding the heavy chain of the antibody, antibody fragment or antibody derivative thereof according to the present disclosure is a nucleic acid sequence of any one of SEQ ID NO: or a functional homologue thereof having nucleic acid sequence which is at least 70 % identical, such as at least 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 95 %, 96 %, 97 %, 98 %, 99 % or 99.9 % identical to any of the variants of SEQ ID NO: 1. In a embodiment, the nucleic acid sequence encoding the heavy chain of the antibody, antibody fragment or antibody derivative thereof according to the present disclosure is a nucleic acid sequence of SEQ ID NO: 1.

[0149] The nucleic acid sequence of the present disclosure may also comprise additional elements than the antibody, antibody fragment or antibody derivative thereof coding region. Such elements are in example, regulatory elements.

[0150] Furthermore, the host cell according to the present disclosure may comprise regulatory elements enabling the controlled over-expression of endogenous or heterologous and / or synthetic nucleic acid sequences.

[0151] The term “regulatory element", comprises promoter sequences, signal sequence, and / or arrays of transcription factor binding sites that affect transcription and / or translation of a nucleic acid sequence operably linked to the regulatory element.

[0152] Regulatory elements are found at transcriptional and post-transcriptional levels and further enable molecular networks at those levels. For example, at the post-transcriptional level, the biochemical signals controlling mRNA stability, translation and subcellular localization are processed by regulatory elements. RNA binding proteins are another class of post-transcriptional regulatory elements and are further classified as sequence elements or structural elements. Specific sequence motifs that may serve as regulatory elements are also associated with mRNA modifications. A variety of DNA regulatory elements are involved in the regulation of gene expression and rely on the biochemical interactions involving DNA, the cellular proteins that make up chromatin, gene activators and repressors, and transcription factors.

[0153] In general, the transcriptional and translational regulatory sequences include, but are not limited to, promoter sequences, ribosomal binding sites, transcriptional start and stop sequences, translational start and stop sequences, binding sites for gene regulators and enhancer sequences.

[0154] Promoters and enhancers are the primary genomic regulatory components of gene expression. Promoters are DNA regions within 1-2 kilobases (kb) of a gene’s transcription start site (TSS); they contain short regulatory elements (DNA motifs) necessary to assemble RNA polymerase transcriptional machinery. However, transcription is often minimal without the contribution of DNA regulatory elements located more distal to the TSS. Such regions, often termed enhancers, are position-independent DNA regulatory elements that interact with site-specific transcription factors to establish cell type identity and regulate gene expression. Enhancers may act independently of their sequence context and at distances of several to many hundreds of kb from their target genes through a process known as looping. Because of these features, it is difficult to identify suitable enhancers and link them to their target genes based on DNA sequence alone. The promoter, together with other transcriptional and translational regulatory nucleic acid sequences (also termed "control sequences") is necessary to express a given gene or group of genes (an operon).

[0155] Identification of suitable promoter sequences that promotes expression of the specific gene of interest is a tedious task, which in many cases require laborious efforts. In relation to the present disclosure regulatory elements may or may not be post-translational regulators or it may or may not be translational regulators.

[0156] Thus, in one embodiment of the present disclosure the regulatory element comprises one or more elements capable of enhancing the expression, i.e. over-expression of the one or more nucleic acid sequence(s) according to the present disclosure.

[0157] The regulatory elements and the nucleic acid sequence encoding an antibody, antibody fragment or antibody derivative thereof may be combined into a single nucleic acid construct.

[0158] One or more embodiments of the invention relates to the nucleic acid construct comprising the nucleic acid sequence according to the invention.

[0159] One or more aspect(s) of the disclosure relates to a host cell comprising one or more nucleic acid sequence(s) according to the present disclosure and / or nucleic acid construct(s) the present disclosure.

[0160] The nucleic acid construct may comprise at least one regulatory element that facilitates the expression of the antibody, antibody fragment or antibody derivative thereof.

[0161] The nucleic acid construct can be a recombinant nucleic acid sequence. By the term “recombinant nucleic acid sequence”, “recombinant gene / nucleic acid / DNA encoding” or "coding nucleic acid sequence" used interchangeably is meant an artificial nucleic acid sequence (i.e. produced in vitro using standard laboratory methods for making nucleic acid sequences) that comprises a set of consecutive, non-overlapping triplets (codons) which is transcribed into mRNA and translated into a protein when under the control of the appropriate control sequences, i.e. a promoter sequence.

[0162] The boundaries of the coding sequence are generally determined by a ribosome binding site located just upstream of the open reading frame at the 5’end of the mRNA, a transcriptional start codon (AUG, GUG or UUG), and a translational stop codon (UAA, UGA or UAG). A coding sequence can include, but is not limited to, genomic DNA, cDNA, synthetic, and recombinant nucleic acid sequences.

[0163] The term "nucleic acid" includes RNA, DNA and cDNA molecules. It is understood that, as a result of the degeneracy of the genetic code, a multitude of nucleic acid sequences encoding a given protein may be produced. The recombinant nucleic sequence may be a coding DNA sequence e.g., a gene, or non-coding DNA sequence e.g., a regulatory DNA, such as a promoter sequence.

[0164] Accordingly, in one exemplified embodiment the disclosure relates to a nucleic acid construct comprising a coding nucleic sequence, i.e. a recombinant DNA sequence encoding an antibody, antibody fragment or derivate thereof, combined with a non-coding regulatory DNA sequence, e.g. a recombinant promoter DNA sequence, or a synthetic promoter sequence, wherein the coding and promoter sequences are operably linked.

[0165] The term “operably linked” refers to a functional relationship between two or more nucleic acid (e.g., DNA) segments. Operably linked refers to the functional relationship of a transcriptional regulatory sequence to a transcribed sequence. For example, a promoter sequence is operably linked to a coding sequence if it stimulates or modulates the transcription of the coding sequence in an appropriate host cell or other expression system.

[0166] Generally, promoter sequences that are operably linked to a transcribed sequence are physically contiguous to the transcribed sequence, i.e., they are c / s-acting.

[0167] In one exemplified embodiment, the nucleic acid construct of the disclosure may be a part of the vector DNA, in another embodiment the construct it is an expression cassette / cartridge that is integrated in the genome of a host cell.

[0168] Accordingly, the term “nucleic acid construct” means an artificially constructed segment of nucleic acid, in particular a DNA segment, which is intended to be 'transplanted' into a target cell, e.g. a mammalian cell, express or to modify expression of a gene / coding DNA sequence that may be included in the construct.

[0169] Integration of the nucleic acid construct of interest comprised in the construct (expression cassette) into the genome of the host cell can be achieved by conventional methods known to the skilled person.

[0170] In general, mammalian cells are preferred for the production of therapeutic antibodies, as they produce antibodies with mammalian glycosylation patterns and generally mammalian cells are better for the production of correctly folded antibody, antibody fragment or antibody derivative thereof.

[0171] In one embodiment, the host cell is a mammalian cell selected from the group consisting of Chinese hamster ovary (CHO) cells, CHO-K1 , CHO-DG44, mouse myeloma (NSO) cells, baby hamster kidney (BHK) cells, and human embryonic kidney lines (HEK293) cells, or an insect cell.

[0172] In a preferred embodiment the host cell is a Chinese hamster ovary (CHO) cell, such as but not limited to CHO-K1 and CHO-DG44. in another embodiment, the host cell is a mouse myeloma (NSO) cell.

[0173] In yet another embodiment baby hamster kidney (BHK) cells.

[0174] In a yet further the host cell is a human embryonic kidney lines (HEK293) cell.

[0175] In a yet further the host cell is an Insect cell.

[0176] Furthermore, as described in the present disclosure, an antibody, antibody fragment or antibody derivative thereof with a humanized glycosylation pattern may be preferred, i.e., glycosylated antibody, antibody fragment or antibody derivative thereof that is fucose deficient.

[0177] In that regard, one or more embodiments relates to a host cell, wherein the cellular fucose glycosylation pathway is modified to reduce the amount of fucose in the glycosylation of said antibody, antibody fragment or antibody derivative thereof. A modified fucose glycosylation pathway may be obtained as disclosed herein, by inclusion of one or more enzymes that modulates the fucose pathway, so that the glycosylation is fucose deficient.

[0178] Accordingly, in one embodiment, the host cell is capable of producing an antibody, antibody fragment or antibody derivative thereof according to the present disclosure, wherein the cellular fucose glycosylation pathway of said host cell is modulated, such that the host cell produces a fucose deficient antibody, antibody fragment or antibody derivative thereof.

[0179] In one or more embodiment(s), the host cell according to the present disclosure, comprises one or more nucleic acid sequence(s) encoding an antibody, antibody fragment or antibody derivative thereof according to the present disclosure.

[0180] In one or more embodiment(s), the host cell according to the present disclosure, comprises one or more nucleic acid constructs comprising at least one nucleic acid sequence encoding an antibody, antibody fragment or antibody derivative thereof according to the present disclosure.

[0181] In that regard, one or more aspect(s) of the present disclosure relates to an antibody, antibody fragment or antibody derivative thereof according to the present disclosure, produced in a host cell according to the present disclosure.

[0182] In one or more embodiment(s) the antibody, antibody fragment or antibody derivative thereof according to the present disclosure is produced in a host cell, wherein the fucose glycosylation pathway is modulated in order to produce a fucose deficient antibody, antibody fragment or antibody derivative thereof.

[0183] One or more embodiments of the invention relates to a host cell comprising the nucleic acid sequence or the nucleic acid construct according to the invention.

[0184] Conjugates, and specifically, Immunoconjugates are antibody, antibody fragment or antibody derivative thereof conjugated (joined) to a second molecule, usually a toxin, radioisotope or label. Such conjugates of the antibody, antibody fragment or antibody derivative thereof are all aspects of the present disclosure.

[0185] Thus, one or more aspect(s) of the present disclosure relates to an antibody, antibody fragment or antibody derivative thereof, drug conjugate that binds to human PTK7 comprising: a) an antibody, antibody fragment or antibody derivative thereof according to the present disclosure, b) a linker and / a chelator, and c) a drug selected from the group consisting of a toxin, a radioisotope, an anticancer drug, a cytotoxic drug and a cytostatic drug.

[0186] Chelating linkers are discussed in the below section regarding radioimmunoconjugates and the chelating linkers described therein are therefore all considered useful for conjugates comprising an antibody, antibody fragment or antibody derivative thereof of the present disclosure connected to or associated with a chelating linker.

[0187] Thus, in one or more embodiment(s), said linker is a chelating linker. In one or more embodiment(s), said linker is a chelator.

[0188] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention and further comprising a linker and / or a chelator.

[0189] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the antibody or antibody fragment is conjugated to a chelator and / or a linker.

[0190] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the chelator and / or a linker binds a drug or radionuclide.

[0191] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the chelator and / or a linker forms a complex with a drug or radionuclide.

[0192] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the chelator is selected as one from the group consisting of p-SCN-bn-DOTA, DOTA-NHS-ester, p-SCN-Bn-TCMC; bifunctional linear chelators like p-SCN-Bn-DTPA and CHX- A"-DTPA.

[0193] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the chelator is p-SCN-Bn-TCMC.

[0194] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the chelator is p-SCN-bn-DOTA. In an embodiment, said linker is a chelating linker selected from the group consisting of p-SCN-bn- DOTA, DOTA-NHS-ester and p-SCN-Bn-TCMC.

[0195] One or more aspects of the invention relates to an antibody-drug conjugate comprising an antibody according to or an antibody fragment according to the invention.

[0196] An immunotoxin is a human-made protein that consists of a targeting portion such as an antibody, linked to a toxin. When the protein binds to that cell, it is taken in through endocytosis or similar pathway, and the toxin kills the cell.

[0197] These immunotoxins are usually used for the treatment of some kinds of cancer and a few viral infections.

[0198] These proteins are usually made of a modified antibody or antibody fragment, attached to a fragment of a toxin.

[0199] The targeting portion is composed of the Fv portion of an antibody that targets a specific cell type. In that sense the targeting portion may be an antibody, antibody fragment or antibody derivative thereof according to the present disclosure.

[0200] The toxin is usually a cytotoxic protein or compound derived from a bacterial or plant protein or of synthetic origin, from which the natural binding domain has been removed so that the Fv directs the toxin to the antigen on the target cell.

[0201] In one or more embodiment(s) the toxin is a chemotherapeutic molecule, including, but not limited to alkylating agents (cisplatin, carboplatin, oxaliplatin, mechlorethamine, cyclophosphamide, chlorambucil, ifosfamide), anti-metabolites (azathioprine, mercaptopurine, pyrimidines), alkaloids (vincristine, vinblastine, cinorelbine, vindesine, paclitaxel, docetaxel, etoposide, teniposide), topoisomerase inhibitors (irinotecan, topotecan, amascrine, etoposide, teniposide) and cytotoxic antibiotics (actinomycin, doxorubicin, daunorubicin, calrubicin, idarubicin, epirubicin, bleomycin, plicamycin, mitomycin).

[0202] In one embodiment doxorubicin is conjugated to the antibody, antibody fragment or antibody derivative thereof via the cross-linker SMCC-hydrazide (4-[N-maleimidomethyl]cyclohexane-1- carboxylhydrazide).

[0203] The immunotoxin works by the antibody (or other targeting moiety) binding to an antigen on the target cell followed by toxin that enters and kills the cell.

[0204] Thus, an aspect of the present disclosure relates to an immunotoxin that comprises antibody, antibody fragment or antibody derivative thereof according to the present disclosure. An aspect of the present disclosure relates to an antibody, antibody fragment or antibody derivative thereof, drug conjugate that binds to human PTK7 according to present invention, coupled or linked to an anticancer drug, a cytotoxic drug, or a cytostatic drug.

[0205] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the antibody-drug conjugate comprises a radionuclide.

[0206] An aspect of the present disclosure relates to a radioimmunoconjugate that binds human PTK7 comprising an antibody, antibody fragment or antibody derivative thereof according to the present disclosure, a linker, and a radionuclide selected from the group consisting of211At,213Bi,212Bi,212Pb,225Ac,227Th,224Ra,223Ra,"Y,161Tb,186Re,188Re,199Au,194lr,166Ho,159Gd,153Sm,149Pm,142Pr,111Ag,109Pd,77As,67Cu,64Cu,47Sc, and177Lu.

[0207] In a preferred embodiment, said drug is a radionuclide, selected from the group consisting of177Lu,225Ac,227Th,212Pb and "Y.

[0208] In another embodiment of the present disclosure the radionuclide is177Lu.

[0209] In another embodiment of the present disclosure the radionuclide is212Pb.

[0210] In yet another embodiment the radionuclide is another beta-emitter or an alpha-emitter.

[0211] The radionuclide may be attached to the antibody by first reacting a bifunctional chelator, e.g., p- SCN-bn-DOTA (Macrocyclics, Tx, USA), with the antibody, followed by purification to remove unconjugated chelator, and then reaction of the chelator antibody conjugate with the radionuclide, followed by purification to remove any unconjugated radionuclide.

[0212] Alternatively, the chelator and the radionuclide can be combined firstly and subsequently conjugated to the antibody.

[0213] Chelating linkers like, e.g., p-SCN-bn-DOTA, can be used for conjugating other metal radionuclides to the antibody, antibody fragment or antibody derivative thereof according to the present disclosure.

[0214] Any type of linker with sufficient complexing ability and a functional group allowing direct or indirect conjugation to a protein or a peptide could be used.

[0215] Examples of such linkers are described in the literature (e.g. Brechbiel, 2008; Liu, 2008). Some useful examples are bifunctional cyclic chelators like p-SCN-bn-DOTA, DOTA-NHS-ester, p-SCN- Bn-TCMC; bifunctional linear chelators like p-SCN-Bn-DTPA and CHX-A"-DTPA.

[0216] The radionuclides in the present disclosure will preferably be conjugated to a targeting molecule by using bifunctional chelators. These could be cyclic, linear or branched chelators. Particular reference may be made to the polyaminopolyacid chelators which comprise a linear, cyclic or branched polyazaalkane backbone with acidic (e.g. carboxyalkyl) groups attached at backbone nitrogens.

[0217] Examples of suitable chelators include DOTA derivatives such as p-isothiocyanatobenzyl-1 ,4,7,10- tetraazacyclododecane-1 ,4,7,10-tetraacetic acid (p-SCN-Bz-DOTA) or S-2-(4- lsothiocyanatobenzyl)-1 ,4,7,10-tetra(2-carbamoylmethyl)cyclododecane and DTPA derivatives such as p-isothiocyanatobenzyl-diethylenetriaminepentaacetic acid (p-SCN-Bz-DTPA), the first being cyclic chelators, the latter linear chelators.

[0218] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the chelator is selected as one from the group consisting of DOTA derived chelators such as: p-SCN-bn-DOTA, DOTA-NHS-ester, p-SCN-Bn- DOTAM (TCMC); bifunctional linear chelators like p-SCN-Bn-DTPA and CHX-A"-DTPA, and further Me-3,2-HOPO or other 3- hydroxy-N-methyl-2-pyridinone derived chelates, Crypt-OH or other derivatives of [2.2.2]-cryptands and H2macropa-NCS or other macropa derived chelates.

[0219] Metallation of the complexing moiety may be performed before or after conjugation of the complexing moiety to the targeting moiety.

[0220] The radiolabeling procedure will in general be more convenient in terms of time used etc if the chelator is conjugated to the antibody before the radiolabeling takes place.

[0221] The principles of preparing radiolabeled conjugates using chelators attached to antibodies are described broader in e.g. Liu, 2008.

[0222] Thus, an antibody, antibody fragment or antibody derivative thereof according to the present disclosure can be used to prepare radioimmunoconjugates with differences in radiation properties and effective half-lives.

[0223] For example anti-PTK7 radioimmunoconjugate consisting of a an antibody comprising a light chain according to SEQ ID NO: 4 and a heavy chain according to SEQ ID NO: 3, a chelating linker and a beta or alpha emitting radionuclide including, but not limited to177Lu,211At,213Bi,212Bi,212Pb,225Ac,227Th, "Y,161Tb,224Ra,223Ra,186Re,188Re,199Au,194lr,166Ho,159Gd,153Sm,149Pm,142Pr,111Ag,109Pd,77As,64Cu,67Cu,47Sc can be prepared and used for preparing pharmaceutical preparations and used in therapeutic applications.

[0224] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention wherein the radionuclide is selected as any one from the group consisting of211At,213Bi,212Bi,212Pb,224Ra,223Ra,225Ac,227Th, "Y,161Tb,186Re,188Re,199Au,194lr,166Ho,159Gd,153Sm,149Pm,142Pr,111Ag,109Pd,77As,67Cu,64Cu,47Sc, and177Lu. One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention comprising a radionuclide, wherein the radionuclide is212Pb.

[0225] One or more embodiments of the invention relates to an antibody-drug conjugate according to the invention comprising a radionuclide, wherein the radionuclide is177Lu.

[0226] An aspect of the present disclosure relates to a positron emitting immunoconjugate that binds human PTK7 comprising an antibody, antibody fragment or antibody derivative thereof according to the present disclosure, a linker, and a positron emitting nuclide selected from the group consisting of11C,13N,15O,18F,64Cu, and89Zr.

[0227] Immunoconjugates may also be used in positron emission tomography.

[0228] In that sense the immunoconjugates for positron imaging are enriched in positron emitting nuclides.

[0229] The position emitting conjugates are usually prepared just prior to the imaging due to the relatively short half-life of the positron emitting nuclides.

[0230] Position emitting conjugates, usually comprises of a targeting molecule conjugated to a compound that is enriched in a positron emitting isotope.

[0231] The positron emitting nuclide may be attached to the antibody by first reacting a bifunctional chelator, e.g., p-SCN-bn-Deferoxamine, (macrocycles, US) with the antibody, followed by purification to remove unconjugated chelator, and then reaction of the chelator antibody conjugate with the positron emitting nuclide, followed by purification to remove any unconjugated positron emitting nuclide.

[0232] Alternatively, the chelator and the positron emitting nuclide can be combined firstly and subsequently conjugated to the antibody.

[0233] Furthermore, the compound conjugated to the targeting molecule may be a compound enriched in positron emitting nuclide may be enriched in11C,13N,15O or18F.

[0234] Chelating linkers like, e.g., p-SCN-Bn-NOTA, can be used for conjugating other metal positron emitting nuclide to an antibody, antibody fragment or antibody derivative thereof in similar fashion to that described for89Zr and64Cu.

[0235] Any type of linker with sufficient complexing ability towards the positron emitting nuclide and a functional group allowing direct or indirect conjugation to a protein or a peptide could be used.

[0236] The positron emitting nuclides of the present disclosure will preferably be conjugated to a targeting molecule by using bifunctional chelators. These could be cyclic, linear or branched chelators. Particular reference may be made to the polyaminopolyacid chelators which comprise a linear, cyclic or branched polyazaalkane backbone with acidic (e.g. carboxyalkyl) groups attached at backbone nitrogens.

[0237] One or more aspects of the invention relates to a composition comprising an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention.

[0238] Antibodies, fragments and derivates thereof are usually applied in the treatment of diseases formulated in pharmaceutical compositions.

[0239] Such compositions are optimized for parameters such as physiological tolerance and shelf-life. Thus, one or more aspect(s) of the present disclosure relates to a pharmaceutical composition comprising, as the active ingredient, one or more antibody / antibodies, antibody fragment(s) or antibody derivative(s) thereof and / or an antibody, antibody fragment or antibody derivative thereof drug conjugate according to the present disclosure, and a pharmaceutically acceptable carrier.

[0240] One or more aspects of the invention relates to a pharmaceutical composition comprising an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelatorantibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention.

[0241] An embodiment of the present disclosure relates to a pharmaceutical composition as described above, further comprising one or more additional therapeutic agents.

[0242] In another embodiment of the present disclosure are said one or more additional therapeutic agents selected from agents that induce apoptosis.

[0243] An immunotherapeutic molecule, such as an antibody, antibody fragment or antibody derivative thereof and / or conjugate thereof as described in the present disclosure, would typically be provided as a pharmaceutical composition potentially consisting of a radionuclide, according to the description above, linked via a chelator to the antibody, antibody fragment or antibody derivative thereof dissolved in a buffer solution, which to a substantial degree maintain the chemical integrity of the immunotherapeutic and / or conjugate thereof and is physiologically acceptable for infusion into patients.

[0244] Thus, an aspect of the present disclosure relates to a pharmaceutical composition comprising a antibody, antibody fragment or antibody derivative thereof according to the present disclosure, and an pharmaceutically acceptable carrier and / or excipient.

[0245] In one or more embodiment(s) of present disclosure relates to a pharmaceutical composition comprising a drug-immunoconjugate of the present disclosure, and a pharmaceutically acceptable carrier and / or excipient.

[0246] In one or more embodiment(s) of present disclosure relates to a pharmaceutical composition comprising a radioimmunoconjugate of the present disclosure, and a pharmaceutically acceptable carrier and / or excipient.

[0247] Acceptable pharmaceutical carriers include but are not limited to non-toxic buffers, fillers, isotonic solutions, etc. More specifically, the pharmaceutical carrier can be but are not limited to normal saline (0.9 %), half-normal saline, Ringer’s lactate, 5 % Dextrose, 3.3 % Dextrose / 0.3 % Saline.

[0248] The physiologically acceptable carrier can contain a radiolytic stabilizer, e.g., ascorbic acid, which protect the integrity of the pharmaceutical during storage and shipment.

[0249] One embodiment of the present disclosure comprises the pharmaceutical composition of the present disclosure and one or more additional antibodies or immunoconjugates.

[0250] One or more aspect(s) of the present disclosure relates to a method for producing an antibody, antibody fragment or antibody derivative thereof according to the present disclosure, the method comprising, a) introducing into a mammalian host cell one or more nucleic acid construct(s) of the present disclosure, b) culturing said host cell in a suitable media, c) recovering said antibody, antibody fragment or antibody derivative thereof from the culturing broth, and d) purifying the antibody, antibody fragment or antibody derivative thereof.

[0251] Thus, one or more aspects of the invention relates to a method for producing an antibody or an antibody fragment according to the invention, comprising providing a host cell comprising the nucleic acid sequence or the nucleic acid construct according to the invention, culturing the host under conditions ensuring expression of the nucleic acid or nucleic acid construct, and recovering the antibody or antibody fragment. In one embodiment of said method, the host cell is a mammalian cell selected from the group consisting of Chinese hamster ovary (CHO) cells, CHO-K1 , CHO-DG44, mouse myeloma (NSO) cells, baby hamster kidney (BHK) cells, and human embryonic kidney lines (HEK293) cells, or an insect cell.

[0252] In one embodiment of said method, the cellular fucose glycosylation pathway of said host cell is modulated, such that the host cell produces a fucose deficient antibody, antibody fragment or antibody derivative thereof.

[0253] In the present disclosure, culturing refers to the process by which cells are grown under controlled conditions, generally outside their natural environment, thus a method used to cultivate, propagate, and grow a large number of cells.

[0254] After culturing of the host cells and expression of the gene product, the purification of the protein is required; but since the vector is introduced to a host cell, the protein of interest should be recovered and / or purified from the proteins of the host cell.

[0255] Therefore, to make the purification process easy, the cloned gene could have a tag. This tag could be histidine (His) tag or any other marker peptide or protein such as but not limited to the Albumin-binding protein. As some embodiment(s) of the present disclosure relates to antibodies, antibody fragments and antibody derivates thereof, such a tag could also be the Fc domain of said antibodies, antibody fragments and antibody derivates thereof. The Fc fragment may be used as a purification tag where an interaction partner such as but not limited to immobilized protein A is used for the purification.

[0256] One or more aspects of the invention relates to a method for producing a chelator-antibody conjugate labelled with a radionuclide according to the invention comprising the steps of obtaining an antibody or antibody fragment according to the invention, mixing said antibody or antibody fragment with a chelator to obtain a chelator antibody conjugate, admixing said chelator-antibody conjugate or chelator antibody fragment conjugate with a radionuclide, and recovering a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate.

[0257] One or more aspect(s) of the present disclosure relates to a method of depleting PTK7 expressing cells from a population of cells, comprising administering to said population of cells, an antibody, antibody fragment or antibody derivative thereof, an antibody, antibody fragment or antibody derivative thereof, drug conjugate, or a pharmaceutical composition according to the present disclosure. One or more aspect(s) of the present disclosure relates to a method of treating disease, wherein targeting of PTK7 expressing cells can provide an inhibition and / or amelioration of said disease, comprising administering a therapeutically effective amount of an antibody, antibody fragment or antibody derivative thereof, an antibody, antibody fragment or antibody derivative thereof, drug conjugate, or a pharmaceutical composition according to the present disclosure.

[0258] Other uses could be treatment of autoimmune diseases and treatment of transplantation related effects.

[0259] One or more aspect(s) of the present disclosure relates to a method of treating cancer and / or inflammatory disease(s) and / or autoimmune disease(s) comprising administering a therapeutically effective amount of an antibody, antibody fragment or antibody derivative thereof, an antibody, antibody fragment or antibody derivative thereof, drug conjugate, or a pharmaceutical composition according to the present disclosure.

[0260] One or more aspect(s) of the present disclosure relates to a method of treating cancer comprising administering a therapeutically effective amount of an antibody, antibody fragment or antibody derivative thereof, an antibody, antibody fragment or antibody derivative thereof, drug conjugate, or a pharmaceutical composition according to the present disclosure.

[0261] One or more aspect(s) the present disclosure relates to the use of an antibody, antibody fragment or antibody derivative thereof, an antibody, antibody fragment or antibody derivative thereof, drug conjugate, or a pharmaceutical composition according to the present disclosure, in inhibiting cancer and / or inflammatory disease(s) and / or autoimmune diseases.

[0262] One or more aspect(s) of the present disclosure relates to the use of an antibody, antibody fragment or antibody derivative thereof, an antibody, antibody fragment or antibody derivative thereof drug conjugate or a pharmaceutical composition according to the present disclosure, in ameliorating cancer and / or inflammatory disease(s) and / or autoimmune diseases.

[0263] The therapy could be based on, but are not limited to, immunotherapy, beta-particle-radiation or alpha-particle-radiation or a combination of these.

[0264] The therapy could be administered either as a monotherapy or in combination with other therapies, preferentially standard treatments. Such other therapies may be pretreatment, surgery, chemotherapy (including doxorubicin, vinblastin and gemcitabine), immunotherapy, photodynamic therapy, proteasome inhibitor (including bortezomib), histone deacetylase inhibitors (including vorinostat and suberoylanilide hydroxamic acid), vitamin D3 and vitamin D3 analogs, cell cycle checkpoint inhibitors (including UCN-01 and 2-(4-(4-Chlorophenoxy)phenyl)-1H-benzimidazole-5- carboxamide), hypoxic cell radiosensitizers (including metronidazole and misonidazole), apoptosis inducers (including withaferin A) radiosensitizers, radioimmunotherapy or a combination of two or more of these.

[0265] By administered is meant intravenous infusion or intravenous injection. More specifically, the pharmaceutical composition of the present disclosure can be administered directly in a vein by a peripheral cannula connected to a drip chamber that prevents air embolism and allows an estimate of flow rate into the patient.

[0266] In one embodiment of the present disclosure are the uses and methods of treatment of the present disclosure performed in vitro or ex vivo.

[0267] In one embodiment said formulation is suitable for administration by one or more administration routes selected from the group consisting of oral, topical, intravenous, intramuscular, and subcutaneous administration.

[0268] In one embodiment, the amount of the antibody fragment or antibody derivative thereof, or the antibody fragment or antibody derivative thereof, drug conjugate according to the present disclosure is at least 0.1 mg and not more than 1 g.

[0269] In an embodiment of the present disclosure the antibody, antibody fragment or antibody derivate thereof or conjugate thereof dosing is 1-1000 mg per patient, more preferably 5-50 mg per patient.

[0270] In an embodiment of the present disclosure the radioimmunoconjugate dosing is 1-1000 mg per patient, more preferably 5-50 mg per patient, and177Lu amounting to 1 - 200 MBq / kg, more preferably 10-100 MBq / kg of bodyweight.

[0271] In one embodiment the antibody, antibody fragment or antibody derivate thereof or conjugates thereof can be administered in a repeated fashion.

[0272] In another embodiment of the present disclosure the antibody, antibody fragment or antibody derivate thereof or conjugates thereof, according to the present disclosure could be administered in a repeated fashion but with different conjugates, such as but not limited to radionuclides, e.g., beta-radioimmunotherapy could be followed by alpha-radioimmunotherapy or chemo- immunoconjugates or vice versa.

[0273] An embodiment of the present disclosure relates to the use of the antibody, antibody fragment or antibody derivate thereof or conjugates thereof, of the present disclosure administered in combination with or in addition to other therapy.

[0274] In an embodiment of the present disclosure the other therapies are selected from pretreatment, chemotherapy, monoclonal antibody therapy, surgery, radiotherapy, radioimmunotherapy, and / or photodynamic therapy. In another embodiment of the present disclosure the other therapies are bone marrow transplantation or stem cell transplantation and / or therapy.

[0275] One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use as a medicament.

[0276] One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of cancer.

[0277] One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of ovarian cancer.One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of cancer.

[0278] One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of a PTK7 expressing cancer from but not limited to ovarian cancer, cervical cancer, endometrial cancer, breast cancer including triple-negative breast cancer, renal cancer, bladder cancer including urothelial cancer, colorectal cancer, mesothelioma, osteosarcoma, brain cancer, including glioma, head and neck cancer, liver cancer including hepatocellular carcinoma, lung cancer including nonsmall-cell lung cancer, lymphoma, myeloma, leukemia, melanoma, neuroendocrine cancer including carcinoid, pancreatic cancer, prostate cancer, testis cancer, skin cancer, gastric cancer, squamous cell carcinoma including esophageal squamous cell carcinoma, and thyroid cancer..

[0279] One or more embodiments of the present invention relates an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of ovarian cancer.

[0280] One or more embodiments of the present invention relates an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of mesothelioma.

[0281] One or more embodiments of the present invention relates an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of bladder cancer.

[0282] One or more embodiments of the present invention relates an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of glioma.

[0283] One or more embodiments of the present invention relates an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of osteosarcoma.

[0284] One or more embodiments of the present invention relates an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of breast cancer.

[0285] One or more embodiments of the present invention relates an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the treatment of colorectal cancer,

[0286] In one embodiment said medicament is administered once or sequential.

[0287] One or more aspect(s) of the present disclosure relates to an antibody, antibody fragment or antibody derivative thereof, conjugate that binds to human PTK7 comprising: a) an antibody, antibody fragment or antibody derivative thereof according to the present disclosure, b) a linker and / or chelator, and c) a compound enriched in one or more isotopes selected from the group consisting of 11C,13N,15O,18F, and89Zr.

[0288] One or more aspect(s) of the present disclosure relates to an antibody, antibody fragment or antibody derivative thereof conjugate according to the present disclosure, for use in positron emission tomography imaging.

[0289] In one embodiment, said imaging is for providing diagnosis, staging, and monitoring treatment of cancers.

[0290] One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use in the diagnosis, monitoring or imaging of cancer.

[0291] One or more aspects of the invention relates to an antibody according to the invention, an antibody fragment according to the invention, an antibody drug conjugate according to the invention, a nucleic acid sequence according to the invention, a nucleic acid construct according to the invention, a host cell according to the invention, an antibody or antibody fragment produced according to the invention, or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced according to the invention, or composition or pharmaceutical composition according to the invention for use as an imaging agent.

[0292] One or more aspect(s) of the present disclosure relates to a kit for the production of an antibody fragment or antibody derivative thereof, drug conjugate according to the present disclosure comprising, a) two or more vials, wherein one vial contains a conjugate comprising a drug linked to a linker, and one vial comprising an antibody, antibody fragment or antibody derivative thereof according to the present disclosure, and b) optionally instructions for preparing said antibody-drug conjugate.

[0293] In the present disclosure “radioimmunoconjugate” and “radionuclide conjugate” are used interchangeably.

[0294] One or more aspect(s) of the present disclosure relates a kit for the production of an antibody fragment or antibody derivative thereof, radionuclide or positron emitting nuclide conjugate, according to the present disclosure comprising, a) two or more vials, wherein one vial contains a conjugate comprising a chelator linked to an antibody fragment or antibody derivative thereof according to the present disclosure, a second vial containing a radionuclide or positron emitting nuclide, and b) optionally, instructions for preparing said radioimmunoconjugate, or positron emitting nuclide-immunoconjugate.

[0295] One or more aspect(s) of the present disclosure relates a kit for the production of an antibody fragment or antibody derivative thereof, radioimmunoconjugate according to the present disclosure comprising, a) two or more vials, wherein one vial contains a conjugate comprising a chelator linked to an antibody fragment or antibody derivative thereof according to the present disclosure, a second vial containing a radionuclide, and b) optionally, instructions for preparing said radioimmunoconjugate conjugate.

[0296] Thus, one or more aspects of the invention relates to kit for producing a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate, the kit comprising an antibody or an antibody fragment according to the invention, a solution of a radionuclide selected as any one of the group consisting of211At,213Bi,212Bi,212Pb,225Ac,227Th, "Y, 161Tb >186Re, 224Ra223Ra188Rei199^, 194|r, 166|_|O159^ , 153S m, 149pm, 142pr, 111Ag, 109pd,77As,67Cu,64Cu,47Sc, and177Lu, and optionally instructions for producing the radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate using the materials provided in the kit.

[0297] One or more aspects of the invention relates to kit for producing a radionuclide labelled chelatorantibody conjugate or radionuclide labelled chelator-antibody fragment conjugate, the kit comprising a chelator-antibody conjugate or complex, wherein the antibody is an antibody or an antibody fragment according to the invention, a solution of a radionuclide selected as any one of the group consisting of211At,224Ra,223Ra,213Bi,212Bi,212Pb,225Ac,227Th,90Y,161Tb,186Re,188Re,199Au,194lr,166Ho,159Gd,153Sm,149Pm,142Pr,111Ag,109Pd,77As,67Cu,64Cu,47Sc, and177Lu, and optionally instructions for producing the radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate using the materials provided in the kit.

[0298] A kit, according to the present disclosure may require some procedures to be performed, e.g., radiolabeling and / or purification to take place before infusion.

[0299] An embodiment of the present disclosure relates to a kit of the present disclosure, wherein the content of one or several of the vials are either lyophilized or in a solution. By mixing the contents of the two vials to generate the drug-immunoconjugate or radioimmunoconjugate the final product will appear. Thus, in another embodiment of the present disclosure the radioimmunoconjugate is generated by mixing the content of the two vials.

[0300] This product may need purification prior to use.

[0301] One or more embodiments of the invention relates to a kit according to the invention, wherein the chelator is selected as any one of p-SCN-bn-DOTA and p-SCN-Bn-TCMC and the radionuclide is selected as any one of177Lu or212Pb.

[0302] One or more embodiments of the invention relates to a kit according to the invention, wherein the chelator is selected as TCMC and the radionuclide is selected as177Lu.

[0303] One or more embodiments of the invention relates to a kit according to the invention, wherein the chelator is selected as Bn-DOTA and the radionuclide is selected as212Pb.

[0304] It should be understood that any feature and / or aspect discussed above in connections with the compounds according to the invention apply by analogy to the methods described herein.

[0305] The following figures and examples are provided below to illustrate the present invention. They are intended to be illustrative and are not to be construed as limiting in any way.

[0306] BRIEF DESCRIPTION OF THE FIGURES

[0307] Figure 1

[0308] Figure 1 shows flow cytometry data showing binding of the 01-1 antibody to A) a human ovarian cancer cell line (Skov-3), B) a human osteosarcoma cell line (OHS) and C) a human glioblastoma cell line T98G. Anti-mouse secondary control histograms are shown in black (Sec.ctr) and cells with positive 01-1 binding in grey (01-1)

[0309] Figure 2

[0310] Figure 2 shows immunohistochemistry analysis of 01-1 conjugated to HRP. Positive staining of tumor cells shown to with x10 (top) and x 20 (bottom) magnificatrion.

[0311] Figure 3

[0312] Figure 3 Shows internalization of the murine 01-1 antibody in the human ovarian cancer cell line A2780 overtime. Control samples are untreated cells and a non-binding murine IgG 1 isotype control. Figure 4

[0313] Figure 4 shows biodistribution of 177Lu-OI-1 in mice bearing subcutaneous OHS xenografts. The data are presented as percentage of injected dose per gram tissue at different time points after injection, with error bars corresponding to the standard error of the mean.

[0314] Figure 5

[0315] Figure 5 shows flow cytometry data showing binding of the chimeric 01-1 (chOI-1) to A) a human osteosarcoma cancer cell line (OHS) and B) a human ovarian cancer cell line (Skov-3). Anti-human secondary control histograms are shown in black (Sec.ctr) and cells with chimeric 01-1 binding in grey (chOI-1).

[0316] Figure 6

[0317] Figure 6 shows weight of intraperitoneal SKOV-3 tumors harvested from mice, 49-50 days after a single dose treatment of either saline, ChOI-1 , 180 kBq 212Pb-ChOI-1 or 405 kBq 212Pb-ChOI-1. N= 8 mice per group. Data shown represents the mean ± SD. Statistical significance (p < 0.005) determined by one way ANOVA and the Tukey post hoc test.

[0318] Figure 7

[0319] Figure 7 A and B show saturation curves of PTK7-bound chOI-1-Alexa488 on human bladder cancer cells. Data are plotted as hyperbolic curves and demonstrate an equilibrium dissociation constant (KD) for PTK7-bound chOI-1-Alexa488 as; A) 3.6 x 10-8M for KU-19-19 cells. B) 2.6 x 108M for 647-V cells. X-axis represents concentration (nM) of chOI-1-Alexa488 antibody, and Y-axis represents calculated relative Mean Fluorescence Intensity (rMFI) based on MFI of unstained cells. Figure 7 C shows internalization of PTK7-bound chOI-1-Alexa488 antibody within KU-19-19 cells. Line graph presenting rMFI of Alexa488 fluorescence for each sample during a 24-hour time period. X-axis represents elapsed time of antibody incubation in hours, and Y-axis represents calculated rMFI based on MFI of unstained cells. The provided data is a representative of at least two independent experiments, in which each point represents mean of samples ± 1SD.

[0320] EXAMPLES

[0321] Example 1 - Immunization, fusion and selection of antibody producing hybhdoma clone

[0322] Animals were immunized repeatedly with human osteosarcoma cells and extracts of cellular membrane proteins. Spleen cells from the immunized mice were fused with myeloma cells according to standard procedure for generation of hybhdoma cells. Supernatants of all growing hybridoma clones of the fusions were tested by flow cytometry for binding to cancer cell lines. (Example 3).

[0323] Supernatants of positive clones were further selected by verifying that the antibodies showed no binding to isolations of human peripheral blood mononuclear cells (PBMC) (Example 3).

[0324] Selected clones were propagated through repeated expansion and recloning and purification and tested on a broader panel of cancer cell lines by flow cytometry as well as PBMC from additional individuals (described in Example 3).

[0325] One of the clones, 01-1 , was found to bind to osteosarcoma cell lines and were subcloned and retested until a clone yielding relevant immune reactivity was identified. This was expanded and stocks were collected and frozen in liquid nitrogen.

[0326] Example 2 - Sequence identity of the variable chains of the 01-1 murine monoclonal antibody

[0327] The coding DNA sequences of the variable heavy (VH) and light (VL) chain of the 01-1 mAb produced by hybridoma clone m4_10 (original annotation) was determined by sequencing of RNA from hybridoma subclone m4_10_2_12.

[0328] Total RNA was extracted from frozen hybridoma cells and reverse transcribed into cDNA using universal primers. RACE was then performed on the cDNA to amplify the variable antibody fragments. Ten clones for each variable fragment were sequenced to confirm the identity.

[0329] The nucleic acid sequences encoding the variable heavy chain and the light heavy chain are SEQ ID NO: 1 and SEQ ID NO: 2, respectively.

[0330] The amino acid sequences corresponding to the variable heavy chain and the light heavy chain are SEQ ID NO: 3 and SEQ ID NO: 4, respectively.

[0331] Thus, the variable heavy chain can be found in SEQ ID NOs 1 and 3, while the variable light chain can be found in SEQ ID NOs 2 and 4.

[0332] Heavy chain: DNA sequence (414 bp)

[0333] Leader sequence-FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 (SEQ ID NO: 1)

[0334] ATGGAATGGTGCTGGGTCTTTCTCTTCCTCCTGTCAGTAACTGCAGGTGTCCACTCCAAG GTCCACCTGCAGCAGTCTGGAGCTGAACTGGTGAAACCCGGGGCATCAGTGAAGCTGTCC TGCAGGGCTTCTGGCTACAGTTTCACTGAGTATATTATACACTGGGTAAAGCAGAGGTCT GGACAGGGTCTTGAGTGGATTGGGTGGTTTTACCCTGGAAGTGGAACTATAAAGTACAAT GAGAAATTCAAGGACAAGGCCACATTGACTGCGGACAAATCCTCCAGCACAGTCTATATG GAAC T T AG T AGAT T GAG AT C T GAAGAC TCTGCGGTCTATTTCTGTG C AAGAC AC GAAAT A TATGGGGCCTCGTTTGCTTACTGGGGCCAAGGAACTCTGGTCACTGTCTCTGCA The heavy chain DNA CDR1 (CDRH1 ) sequence is:

[0335] GCTACAGTTTCACTGAGTATATTATACAC ( SEQ ID NO : 7 )

[0336] The heavy chain DNA CDR2 (CDRH2) sequence is:

[0337] TGGTTTTACCCTGGAAGTGGAACTATAAAGTACAATGAGAAATTCAAGGAC ( SEQ ID

[0338] NO : 8 )

[0339] The heavy chain DNA CDR3 (CDRH3) sequence is:

[0340] CACGAAATATATGGGGCCTCGTTTGCTTAC ( SEQ ID NO : 9 )

[0341] Heavy chain: Annino acids sequence (138 AA) (SEQ ID NO: 3)

[0342] Leader sequence-FR1 -CDR1 -FR2-CDR2-FR3-CDR3-FR4

[0343] MEWCWVFLFLLSVTAGVHSKVHLQQSGAELVKPGASVKLSCRASGYSFTEYI IHWVKQRS GQGLEWIGWFYPGSGTIKYNEKFKDKATLTADKSSSTVYMELSRLTSEDSAVYFCARHEI YGAS FAYWGQGTLVTVSA

[0344] The heavy chain amino acid CDR1 (CDRH1) sequence is:

[0345] GYSFTEYI IH ( SEQ ID NO : 10 )

[0346] The heavy chain amino acid CDR2 (CDRH2) sequence is:

[0347] WFYPGSGTIKYNEKFKD ( SEQ ID NO : 11 )

[0348] The heavy chain amino acid CDR3 (CDRH3) sequence is:

[0349] HEIYGASFAY ( SEQ ID NO : 12 )

[0350] Light chain: DNA sequence (381 bp) (SEQ ID NO: 2)

[0351] Leader sequence-FR1 -CDR1 -FR2-CDR2-FR3-CDR3-FR4

[0352] ATGAAGTCACAGACCCAGGTCTTCGTATTTCTACTGCTCTGTGTGTCTGGTATTCATGGG

[0353] AATATTGTGATGACCCAGACTCCCAAATTCCTCCTTGTATCAGCAGGGGACAGGGTTACC

[0354] ATAACCTGCAAGGCCAGTCAGAGTGTTAATAATGATGTAACTTGGTACCAACAGAGGCCA

[0355] GGGCAGTCTCCTAAATTACTGATATACTATGCTTCCAATCGCTACACTGGAGTCCCTGAT

[0356] CGTTTCACTGGCAGTGGATATGGGACGGATTTCACTTTCACCATCAGCACTGTGCAGGCT

[0357] GAAGACCTGGCAATTTATTTCTGTCAGCAGGATTATAGCCCCCCGTACACGTTCGGAGGG

[0358] GGGACCAAGCTGGAAATAATA

[0359] The light chain DNA CDR1 (CDRL1) sequence is: AAGGCCAGTCAGAGTGTTAATAATGATGTAACT ( SEQ ID NO : 13 )

[0360] The light chain DNA CDR2 (CDRL2) sequence is:

[0361] TATGCTTCCAATCGCTACACT ( SEQ ID NO : 14 )

[0362] The light chain DNA CDR3 (CDRL3) sequence is:

[0363] CAGCAGGATTATAGCCCCCCGTACACG ( SEQ ID NO : 15 )

[0364] Light chain: Annino acids sequence (127 AA) (SEQ ID NO: 4)

[0365] Leader sequence-FR1 -CDR1 -FR2-CDR2-FR3-CDR3-FR4

[0366] MKSQTQVFVFLLLCVSGIHGNIVMTQTPKFLLVSAGDRVTITCKASQSVNNDVTWYQQRP GQSPKLLIYYASNRYTGVPDRFTGSGYGTDFTFTISTVQAEDLAIYFCQQDYSPPYTFGG GTKLEI I

[0367] The light chain amino acid CDR1 (CDRL1) sequence is:

[0368] KASQSVNNDVT ( SEQ ID NO : 16 )

[0369] The light chain amino acid CDR2 (CDRL2) sequence is:

[0370] YASNRYT ( SEQ ID NO : 17 )

[0371] The light chain amino acid CDR3 (CDRL3) sequence is:

[0372] QQDYSPPYT ( SEQ ID NO : 18 )

[0373] Example 3 - Flow Cytometric analyses and Immunohistochemistry

[0374] Different flow cytometry screens were performed on the 01-1 antibody for initial characterization of the suitability of the antibody for therapeutic development.

[0375] Example 3A Preparation of cells and flow cytometry:

[0376] Human tumor cell lines were grown in DMEM or RPMI Gibco (Invitrogen), supplemented with 10 % FCS, and with extra glutamine (Glutamax) and antibiotics (Pen / Strep) dependent on the recommendation of the supplier. Cells were grown in a humid atmosphere with 95 % air and 5 % CO2. Adherent cell lines were harvested at 80-100 % confluence by 2 x wash in DPBS followed by trypsination (TrypLE express, Invitrogen). Cells grown in suspension were harvested by centrifugation.

[0377] Peripheral blood mononuclear cell (PBMC) from three individuals were analysed. Whole blood drawn on EDTA glass from healthy donors was diluted 1 :1 with DPBS (DPBS, Hyclone, Thermo Scientific, USA) in a 50 ml tube. Diluted whole blood was layered on top of Lymphoprep (Medinor, Norway) in a ratio 2:1 in a 50 ml tube and centrifuged at 1000*g for 20 min with slow braking. The mononuclear cells from the interface are aspirated and washed twice with DPBS (300*g, 10 min). The pelleted cells are gently resuspended in culture medium (RPMI-1640 with 10% heat- inactivated fetal bovine serum) using a pipette.

[0378] Cell numbers were determined by counting aliquots on a Countess cell counter (Invitrogen) followed by dilution in flow buffer (PBS / DPBS w / 0.5 % BSA). 200.000 to 1 000 000 cells pr sample well was distributed in 5 ml sample tubes or in a 96 well plate (Becton Dickinson PRO-BIND 96). Primary antibody was added on a concentration of 0.01-1 ug / ml and were incubated at 4 °C for 60 min before wash with 200 pl flow buffer.

[0379] Secondary antibody (anti-mouse or anti-human IgG FITC or Alexa488 (was added and incubated for 30 min and washed as in previous step. All wash steps were performed by centrifugation at 1 200 rpm in 5 min. Washed cell pellets were dissolved in 150 ul flow buffer and analysed on FACS Calibur or a CytExpert version 2.4 (Beckman Coulter).

[0380] Example 3B: Determination of IgG subclass for the OI-1 antibody

[0381] Flow cytometry analysis with fluorochrome conjugated murine isotype specific secondary antibodies (lgG1 , lgG2a, lgG2b, lgG3, IgM) were performed. OI-1 was determined to be subclass lgG1.

[0382] Example 3C Binding of OI-1 to fresh preparations of human PBMC

[0383] Flow cytometry analysis of Peripheral Blood Mononuclear Cells (PBMC) from three different individuals confirmed that OI-1 does not bind to human PBMC, supporting that the antibody could be relevant for development for anticancer therapy.

[0384] Example 3D OI-1 binding to human cancer cell lines

[0385] Protein tyrosine kinase 7 (PTK7) is identified as a highly expressed antigen enriched in cancer cells from multiple solid tumors, including but not limited to ovarian cancer, cervical cancer, endometrial cancer, breast cancer including triple-negative breast cancer, renal cancer, bladder cancer including urothelial cancer, colorectal cancer, mesothelioma, osteosarcoma, brain cancer, including glioma, head and neck cancer, liver cancer including hepatocellular carcinoma, lung cancer including non-small-cell lung cancer, lymphoma, myeloma, leukemia, melanoma, neuroendocrine cancer including carcinoid, pancreatic cancer, prostate cancer, testis cancer, skin cancer, gastric cancer, squamous cell carcinoma including esophageal squamous cell carcinoma, and thyroid cancer. The binding of OI-1 to various human cancer cell lines have been confirmed by flow cytometric analysis. OI-1 binds to the glioblastoma cell lines A172, T98G, LN-18 and M059K, to the mesothelioma cell lines MSTO-211H and NCI-H226, to osteosarcoma cell lines OHS, KPDX and Saos-2, and to the ovarian cancer cell lines Skov-3 and Ovcar-3 (Table 1 ). Selected flow cytometry histograms are shown in Figure 1.

[0386] Table 1. Measured binding of 01-1 antibody to a selection of human tumor cell lines by flow cytometric analysis

[0387] Example 3E 01-1 binding to human osteosarcoma tissue by immunohistochemistry

[0388] Procedures for immunohistochemistry (IHC) staining of frozen tissue sections: Biopsy of human osteosarcoma tissues were snap frozen. 5 pM sections of frozen tissue were air dried for one hour before being fixed in acetone at room temperature for 10 minutes. Slides were then dried at room temperature for 10 min. After 2 x 5 min. washes in 0.5 M Tris, the endogenous peroxidase activity was blocked using 3% H2O2 in methanol for 10 min. Sections were washed with distilled water and then 1 x 5 min with 0.5 M Tris. Sections were blocked with 5% BSA in PBS for 20 min. The slides were then incubated with horseradish peroxidase (HRP) conjugated OI-1 (10 pg / ml) in 2.5% BSA in for 60 min at room temperature. Sections were washed 2 X 5 min with 0.5M Tris, and then incubated with liquid DAB and Chromogen system (Dako K3468) for 5 min; 1 drop of DAB + Chromogen in 1 ml of substrate buffer (supplied by Dako). Sections were rinsed in distilled water, counterstained with hematoxylin for 1-2 min, following removal of the excess staining with . 1% HCI in EtOH and rinsing in cold tap water for 10 min. Sections were dehydrated in EtOH, then in xylene and mounted in Perfix.

[0389] Positive staining with tumor cells of tissue samples from human osteosarcoma samples shows that OI-1 has suitable properties for the detection of cancer cells in immunohistochemistry analyses. An example of moderately strong positive staining of osteosarcoma tumor cells is shown in Figure 2.

[0390] Example 4 - Identification of OI-1 target antigen

[0391] Example 4A - Identification of OI-1 target antigen by screen of a cell library

[0392] Identification of PTK7 as the target of OI-1 was achieved by screening an array of cells expressing a protein library consisting of more than 6000 vectors encoding individual human plasma- membrane and cell surface tethered human secreted proteins as well as 396 human heterodimers (Retrogenix).

[0393] Slides were spotted with the control vector ZsGreenl (quadruplicate spots per slide) and the expression vectors in duplicates and used to reverse-transfect HEK293 cells. Once transfection efficiency was confirmed using the control vector, 20 pg / ml of 01-1 antibody was added to each slide in duplicates. A positive control; 0.2 pg / mL of CTLA4-mFc and negative control; PBS were also screened against the transfected cells. The cells were fixed and binding of OI-1 antibody was detected using a secondary antibody (AlexaFluor 647 labeled anti-mouse IgG H+L) and images analysed using ImageQuant software (GE healthcare, Version 8.2). Positive hits were classified as strong, medium, weak, or very weak via visual inspection.

[0394] A confirmation of the protein ‘hits’ was performed using a similar slide set-up as the initial screening, evaluating cells with and without fixation. Target validation was performed by flow cytometry on cells transfected proteins detected as hits in the prior assays.

[0395] The OI-1 antibody showed specific interactions with PTK7, on cell microarrays treated both before and in the absence of fixation, flow cytometry study confirmed strong fluorescence intensity binding to PTK7.

[0396] Example 4B - Binding affinity of OI-1 antibody to PTK7 by surface plasmon resonance analysis

[0397] Multi-cycle kinetics surface plasmon resonance (SPR) were performed to confirm binding and assess affinity of the OI-1 antibody to PTK7. A his tagged PTK7 antigen was purchased from Sino Biological (Cat. No 19399-H08H), and used as the analyte in a SPR analysis 01-1 captured on a Protein A chip. The analysis was performed on a on a Biacore 8K. A Kd value of 6.86 nM was obtained, supporting that the 01-1 antibody binds to the target PTK7 with an affinity relevant for development as a targeting antibody for imaging or therapy.

[0398] Example 5 - Internalization of 01-1

[0399] Example 5A- Assessment of internalization by microscopy

[0400] Surface binding and internalization of murine 01-1 to human tumor cell lines were assessed by confocal microscopy. 1 x 104OHS cells were incubated with cell medium in 8-microscopic plates. Alexa Fluor 488 labelled 01-1 (AI488-OI-1) was added one day after cell seeding. The cells were incubated for 4 hours. Evaluation of surface bound versus internalized antibody was assessed by removal of surface bound antibody in some wells, by a weak acidic treatment (stripping) before imaging of both stripped and non-stripped sample wells.

[0401] Confocal microscopy imaging confirmed antibody internalization in the cells. Example 5B- Assessment of internalization by live cell imaging

[0402] The internalization of the murine 01-1 antibody was assessed using the Incucyte ® S3 Live Cell Imaging instrument. For this assay, the human ovarian cancer cell line A2780 was used. Additionally, the internalization of an non-binding isotype control (murine lgG1 ) antibody was assessed as a negative control. The 01-1 and mlgG1 antibodies were conjugated to a pH sensitive mouse Fabflour antibody labeling dye in a 1 :3 molar ratio using the manufacturer’s protocol. A2780 cells were seeded in a 96 well plate at a density of 20 x 103cells per well. Test antibodies were added to the cells at a concentration of 4 pg / ml. Readout was performed over a period of 72 hours with measurements taken at time intervals of 15 minutes. Analysis was performed using the Basic Analyzer of the Incucyte software v2022B.

[0403] Gradual internalization of OI-1 in the A2780 human ovarian cell lines was observed from start of incubation with the maximum level reached at approximately 10 hours. The specific internalization was evident in comparison to the low non-specific internalization observed with the negative control, a non-binding murine IgG 1 isotype control (Figure 3).

[0404] Example 6: Radiolabeling of OI-1

[0405] Example 6A. Radiolabeling of Bn-DOTA conjugated OI-1 p-SCN-Bn-DOTA (Macrocyclics) was conjugated to OI-1 according to published procedures. Briefly, OI-1 antibody in carbonate buffer of pH 8.5was added to five-fold molar excess of p-SCN- Bn-DOTA and the reaction vial was mixed gently on a thermomixer (Eppendorf) for 2 hours at room temperature. Unconjugated chelator was separated from DOTA-conjugated 01-1 and the buffer exchanged to 0.9% NaCI using a centrifuge filtering cartridge (Vivaspin 15R, 50 kDa MWCO, Sartorius Stedim Biotech). Thereafter the pH was adjusted to 5.5-6.0 by adding ammonium acetate and cationic177Lu in diluted HCI was added and reacted for 15-45 minutes at 37 °C. The radiolabeled product was purified with a Sephadex PD-10 gel filtration. This resulted in a product of 99% radiochemical purity.

[0406] Example 6B. Radiolabeling of SCN-Bn-TCMC conjugated OI-1

[0407] Similar to Bn-DOTA conjugation, OI-1 antibody in carbonate buffer of pH 8.5 was added to five-fold molar excess of p-SCN-Bn-TCMC (Macrocyclics) and the reaction maintained at room temperature and under programmed mixing for 2 hours. The unconjugated TCMC chelator was separated from the TCMC conjugated 01-1 in 0.9% NaCI using a centrifuge ultrafiltration tube (Vivaspin®, 50 kDa MWCO, Cytiva Life sciences). Thereafter, the TCMC conjugated OI-1 antibody was added to212Pb2+solution of pH 5.5-6.0 (pH adjusted using ammonium acetate). The mixture was reacted for 15-30 minutes at room temperature with programmed mixing on a thermomixer. This resulted in a product of >98% radiochemical purity when measured at equilibrium.

[0408] In conclusion: The 01-1 monoclonal antibody could be radiolabeled with both177Lu and212Pb. This indicates that it may be appropriate for creating antibody-radionuclide conjugates relevant for therapeutic applications etc.

[0409] Example 6C: Immunoreactive fraction with radiolabeled OI-1

[0410] The human osteosarcoma cell lines; Saos-2 and OHS, were used to study binding of radiolabeled OI-1 monoclonal antibody by determining the immunoreactive fraction in one-point binding assays in vitro.

[0411] Single cell suspensions of 50-1 13 x 106cells / ml in Dulbecco’s PBS supplemented with 0.5% BSA was prepared. Duplicate samples of 0.15-0.2 ml cell solution were incubated at room temperature for about 60 min with gentle shaking together with radiolabeled mAb: 0.4-6.3 ng of177Lu-DOTA-OI- 1 and 2 ng of212Pb-TCMC-OI-1. Nonspecific binding was estimated from samples preincubated (15-30 min) with 15-20 pg unlabeled mAb prior to the addition of radiolabeled mAb. After that each tube was counted on a gamma counter to determine total radioactivity, washed tree times with 0.5 ml DPBS / BSA and the cell pellets re-measured to determine the cell bound fraction. The fraction of bound mAb was calculated by dividing the amount of cell-bound activity on the total added radioactivity, and the immunoreactivity was the fraction of bound mAb minus the fraction of unspecific bound mAb.

[0412] Results: The immunoreactive fraction ranged from 51 to 72% with177Lu-DOTA-OI-1 in Saos-2 cells and 40-44 % with212Pb-TCMC-OI-1 in OHS cells. In conclusion, radiolabeled OI-1 displayed a relevant binding in cell binding assays.

[0413] Example 7. Affinity assessment using177Lu-OI-1

[0414] Saturation binding experiments were set up to determine the equilibrium dissociation constant, KD, and the number of specific binding sites, Bmax, for177Lu-OI-1 on Saos-2 (osteosarcoma) and SKOV-3 (ovarian cancer) cells.

[0415] Increasing amounts of177Lu-OI-1 , ranging from 25 ng to 22 pg per ml, were added to 0.1 ml samples of 10 x io6cells / ml in Dulbecco’s PBS with 0.5 % BSA, four sample replicates for each concentration. Incubation with gentle agitation was allowed to proceed for 65-95 min at 4°C. Unspecific binding was determined by letting half of the samples preincubate for 30-60 min in the presence of approximately 100 pg / ml unlabeled mAb, before177Lu-OI-1 was added. Measurements of the total and cell-bound activity were carried out as described for the immunoreactivity assay (Example 6C). Values of KD and Bmax were determined from globally fitting the experimental binding data to the sum of a hyperbolic curve (specific binding) and a straight line (nonspecific binding): where B is the number of antigen sites per cell, [T] is the total concentration of177Lu-OI-1 in nmol / L (nM) and NS is the slope of the nonspecific binding. The fitting was performed with GraphPad Prism version 9.4.1 for Windows (GraphPad Software, La Jolla, CA, USA).

[0416] Table 2 Experimentally determined equilibrium dissociation constant, KD, and number of specific binding sites, Bmax, for177Lu-OI-1 on Saos-2 (osteosarcoma) and SKOV-3 (ovarian cancer) cells. The results are given as a range of two separate experiments for each cell line.

[0417] The KD is within the same range as marketed therapeutic antibodies; Rituximab KD = 8 nM (Rituxan, Prescribing information; Genentech, Inc. Med Guide Revision Date July 2012), Trastuzumab KD = 8.2 nM (Costantini et al., 2007) and Erbitux KD = 0.2 nM (Product Monograph, Bristol-Myers Squibb Canada, December 2012).

[0418] Example 8 Biodistribution of177Lu-OI-1 in nude mice with sub cutaneous osteosarcoma tumors

[0419] The177Lu-OI-1 was administered by tail vein injection to mice bearing osteosarcoma (cell line OHS) xenografts. The dosage was between 500 - 1100 kBq 177Lu on 4 - 7 ug OI-1 per mouse. At different time points post injection (from 1 h up to 14 days) blood was collected by cardiac puncture while the mice were under sevoflurane anesthesia. Immediately after blood sampling, the animals were euthanized by cervical dislocation, before tumor and different tissues (lungs, heart, liver, spleen, kidney, stomach, small intestine, large intestine, femur, muscle, brain and skull) were removed at the autopsy. Each tissue sample was weighed, and the radioactivity measured in a gamma counter (Cobra II auto-gamma detector). Samples of the injectates were used as a reference in the measurement procedures. The decay corrected percentages of the injected dose per gram tissue (% I D / g) were calculated for all time points. A group size of two to six animals was used per time point. Tumors with weight less than 50 mg were excluded from the analyses.

[0420] The distribution of radioactivity in various tissues as a function of time after administration of the 177Lu-labeled 01-1 is shown Figure 4. At all time points, the percentage of injected dose per gram tissue was higher for tumor than any other normal organ. The radioactivity in the tumors increased up to 4 days after injection, whereas the uptake in normal tissues decreased or remained unchanged during the same course of time.

[0421] In conclusion, OI-1 had significant tumor targeting abilities in vivo, which is an essential feature for development of anticancer monoclonal antibodies or conjugates thereof.

[0422] Example 9: Recombinant cloning and testing of a human / mouse chimeric variant of OI-1 (mlgG1), ChOI-1 (hlgG1)

[0423] Example 9A: Recombinant cloning and expression of a chimeric OI-1 antibody, chOI-1.

[0424] Recombinant plasmids were constructed encoding heavy (SEQ ID NO:1 and 3) and light chain (SEQ ID NO:2 and 4) of OI-1 in combination with constant regions of human IgG / K light chain (SEQ ID NO:5) and IgG 1 / K (SEQ ID NO:6). The plasmids were transiently transfected into suspension CHO cell cultures.

[0425] The target antibody, ChOI-1 was captured from the cell culture supernatant by HiTrapTM ProteinA HP 5 ml column. Analysis of antibodies by reduced and non-reduced SDS-PAGE and Western confirmed estimated molecular weight of -55 kDa heavy chain, -25 kDa light chain and -150 kDa full length antibodies, respectively.

[0426] SEQ ID NO:5 hlgG / K light chain constant region

[0427] CGAACTGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGA ACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAG GTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGG ACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAA GTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAG GGGAGAGTGT SEQ ID NO:6 hlgG1 / K heavy chain constant region

[0428] GCCTCCACCAAGGGACCAAGCGTGTTCCCACTGGCTCCATCTAGTAAAAGTACTTCAGGGGG AACCGCCGCTCTGGGATGTCTGGTGAAGGATTATTTCCCAGAGCCCGTGACAGTGAGCTGGA ACTCCGGCGCCCTGACCTCCGGAGTGCACACATTTCCAGCTGTGCTGCAGTCAAGCGGCCTG TACTCTCTGTCCTCTGTGGTGACCGTGCCCAGTTCAAGCCTGGGGACTCAGACCTATATCTGC AACGTGAATCACAAGCCAAGTAATACAAAAGTGGACAAGAAAGTGGAACCCAAGAGCTGTGAT AAAACACATACTTGCCCCCCTTGTCCTGCTCCAGAGCTGCTGGGAGGACCAAGCGTGTTCCT GTTTCCACCCAAGCCTAAAGACACCCTGATGATTTCCAGGACACCCGAAGTGACTTGCGTGGT

[0429] GGTGGACGTGTCTCACGAGGATCCTGAAGTGAAGTTCAACTGGTACGTGGATGGCGTGGAG GTGCATAATGCCAAGACCAAACCCAGGGAGGAACAGTACAATAGCACCTATCGCGTGGTGTC CGTGCTGACAGTGCTGCACCAGGACTGGCTGAACGGCAAAGAGTATAAGTGCAAAGTGAGCA ATAAGGCCCTGCCCGCTCCTATCGAGAAAACAATTAGCAAGGCCAAAGGGCAGCCTCGGGAA CCACAGGTGTACACTCTGCCTCCATCCAGAGACGAGCTGACAAAGAACCAGGTGTCTCTGAC TTGTCTGGTGAAAGGGTTCTATCCTTCAGATATTGCTGTGGAGTGGGAAAGCAATGGACAGCC AGAAAACAATTACAAGACCACACCCCCTGTGCTGGACTCTGATGGAAGTTTCTTTCTGTATTCT

[0430] AAGCTGACTGTGGATAAAAGTCGGTGGCAGCAGGGCAACGTGTTTAGCTGTTCCGTGATGCA TGAGGCCCTGCACAATCATTACACCCAGAAGTCTCTGAGTCTGTCACCTGGCAAA

[0431] Example 9B: Analysis of ChOI-1 binding specificity

[0432] To verify that the chimeric variant of 01-1 has equal binding properties as the murine 01-1 , analyses of binding were performed. Flow cytometry analysis of antibody ChOI-1 confirmed specific binding to several cell lines (including OHS, Skov3 and NCI-H226). Figure 5 shows data for selected cell lines.

[0433] Example 10: Tumor growth inhibition of lead-212 labeled chOI-1 in xenograft mouse model

[0434] The therapeutic effect of radiolabeled chOI-1 was evaluated in a mouse model. Human ovarian cancer xenograft was established in nude mice aged 4-5 weeks by intraperitoneal (i.p.) inoculating 5 x 106of Skov-3 cells. Three days after cell inoculation, mice were randomized into groups and each mouse received an i.p injection of either saline, non-radiolabeled chOI-1 , or a dose of212Pb- chOI-1 with a constant antibody amount of 10 pg. Mice were followed over the course of the study, by monitoring weight every 2-3 days. When control mice reached the predefined humane endpoints corresponding to when the mice displayed significant clinical signs of tumor burden (Day of all visible tumors.

[0435] The results of the study (Figure 6) show that targeting PTK7 with the212Pb-chOI-1 antibody is a promising treatment strategy. Even at the lowest activity dose tested (180 kBq per animal), a significant tumor reduction was observed.

[0436] Example 11. Binding and internalization of chimeric 01-1 (ChOI-1 ) in human bladder cancer cell lines.

[0437] Binding of ChOI-1 (Example 9A) to human bladder cancer cell lines were assessed by flow cytometry (methods as in Example 3). The bladder cancer cell lines examined were KU-19-19, T- 24, CAL-29, 647-V and RT-112, human isotype control antibody (hlgG 1 ) was run as parallel negative control samples. Results are shown in Table 3. The highest expression of PTK7 by chOI- 1 binding was detected on RT-112 with a relative Mean Fluorescence Intensity (rMFI) of 14.1 followed by cell line 647-V, KU-19-19, CAL-29 and T-24.

[0438] Table 3: Relative mean Alexa488 fluorescence intensity of antibodies, chOI-1-Alexa488 and hlgG1-Alexa488 on bladder cancer cells. The bladder cancer cells in an amount of 1 x 106cells / sample were incubated with 1 pg chOI-1-Alexa488 or 1 pg hlgG1-Alexa488 for 30 minutes at 4°C and washed twice before being acquired on the flow instrument. The relative Mean Fluorescence Intensity (rMFI) was calculated based on MFI of unstained cells. The saturation binding of ChOI-1 to bladder cancer cell line KU-19-19 and 627-V were further examined. The equilibrium dissociation constant (KD) to the cell lines were found to be 3.6 x 10-8M for KU-19-19 and 2.6 x 10-8M for 647-V (Figure 7 A and B, respectively).

[0439] To investigate functionality of the ChOI-1 antibody, chOI-1-Alexa488 was examined for internalization within high PTK7 expressing human bladder cancer cell line KU-19-19 by flow cytometry, isotype control hlgG1-Alexa488 was used as non-binding negative control in the assay. Antibody incubation timepoints for KU-19-19 cells with chOI-1-Alexa488 were 0.25-, 2-, 6- and 24 h. A persistent increase of Alexa488 signal in trypsin-treated cells was detected at 2 h, 6 h and 24 h incubation, in which the signal was approximately 51.0 ± 6.0 %, 63.0 ± 16.3% and 93.1 ± 9.8 %, respectively, suggesting a subsequent increase in chOI-1-Alexa488 internalization. As a result, chOI-1 antibody bound to surface expressed PTK7 internalizes within KU-19-19 human bladder cancer cells starting with 0.25 h and reaching approximately 93 % at 24 h.

[0440] The flow cytometry assays results confirm binding properties and cellular internalization of ChOI-1 to human bladder cancer cells, that support antibody characteristics of relevance for development as a targeted cancer therapeutics.

Claims

Claims1. An antibody comprising a heavy chain and a light chain, wherein the heavy chain comprises a CDRH1 having an amino acid sequence that is 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 10, a CDRH2 having an amino acid sequence that is 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 11 , and a CDRH3 having an amino acid sequence that is 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 12, and the light chain comprises a CDRL1 having an amino acid sequence that is 90%, such as for example 93, 95, 98, or 99% identical to any one of SEQ ID NO: 16, a CDRL2 having an amino acid sequence that is 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 17, and a CDRL3 having an amino acid sequence that is 90%, such as for example 93, 95, 98 , 99 or 100% identical to any one of SEQ ID NO: 18.

2. The antibody according to any one of the proceeding claims, wherein the heavy chain comprises a CDRH1 having an amino acid sequence according to any one of SEQ ID NO: 10, a CDRH2 having an amino acid sequence according to any one of SEQ ID NO: 11 , and a CDRH3 having an amino acid sequence according to any one of SEQ ID NO: 12, and the light chain comprises a CDRL1 having an amino acid sequence according to any one of SEQ ID NO: 16, a CDRL2 having an amino acid sequence according to any one of SEQ ID NO: 17, and a CDRL3 having an amino acid sequence according to any one of SEQ ID NO: 18.

3. The antibody according to any one of the preceding claims, wherein-the heavy chain comprises an amino acid sequence according to any one of SEQ ID NO: 3, and-the light chain comprises an amino acid sequence according to SEQ ID NO: 4.

4. The antibody according to any one of the preceding claims, wherein- the heavy chain has an amino acid sequence according to SEQ ID NO: 3, and-the light chain has an amino acid sequence according to SEQ ID NO: 4.

5. An antibody fragment of an antibody as defined in any one of the preceding claims, wherein the antibody fragment is an Fv, scFv, Fab, F(ab')2, Fab', or scFv-Fc antibody fragment.

6. An antibody-drug conjugate comprising an antibody according to any one of claims 1-5 or an antibody fragment according to claim 5.

7. The antibody-drug conjugate according to claim 6 further comprising a linker and / or a chelator.

8. The antibody-drug conjugate according to claim 7, wherein the antibody or antibody fragment is conjugated to a chelator and / or a linker.

9. The antibody-drug conjugate according to claim 8, wherein the chelator and / or a linker binds a drug or radionuclide.

10. The antibody-drug conjugate according to claim 9, wherein the chelator and / or a linker forms a complex with a drug or radionuclide.11 . The antibody-drug conjugate according to any one of claims 7-10, wherein the chelator is selected as one from the group consisting of p-SCN-bn-DOTA, DOTA-NHS-ester, p-SCN- Bn-TCMC; bifunctional linear chelators like p-SCN-Bn-DTPA and CHX-A"-DTPA.

12. The antibody-drug conjugate according to any one of claims 7-1 1 , wherein the chelator is p-SCN-Bn-TCMC.

13. The antibody-drug conjugate according to any one of claims 7-1 1 , wherein the chelator is p-SCN-bn-DOTA.

14. The antibody-drug conjugate according to claim 7-13, wherein the antibody-drug conjugate comprises a radionuclide.

15. The antibody-drug conjugate according to claim 7-14, wherein the radionuclide is selected as any one from the group consisting of211At,213Bi,212Bi,212Pb,225Ac,227Th,90Y,161Tb, 186Re,188Re,224Ra,223Ra,199Au,194lr,166Ho,159Gd,153Sm,149Pm,142Pr,111Ag,109Pd,77As, 67Cu,64Cu,47Sc, and177Lu.

16. The antibody-drug conjugate according to any one of claims 7-15, wherein the radionuclide is212Pb.

17. The antibody-drug conjugate according to any one of claims 7-15, wherein the radionuclide is177Lu.

18. A nucleic acid sequence encoding the antibody according to any one of claims 1-4 or the antibody fragment according to claim 5.

19. The nucleic acid construct comprising the nucleic acid sequence according to claim 18.

20. A host cell comprising the nucleic acid sequence according to claim 18 or the nucleic acid construct according to claim 19.21 . A method for producing an antibody according to claims 1-4 or an antibody fragment according to claim 5, comprising providing a host cell comprising the nucleic acid sequence according to claim 18 or the nucleic acid construct according to claim 19,culturing the host under conditions ensuring expression of the nucleic acid or nucleic acid construct, and recovering the antibody or antibody fragment.

22. A method for producing a chelator-antibody conjugate labelled with a radionuclide according to any one of claims 14-19 comprising the steps obtaining an antibody according to any one of claims 1-4, or antibody fragment according to claim 5, mixing said antibody or antibody fragment with a chelator to obtain a chelator antibody conjugate, admixing said chelator-antibody conjugate or chelator antibody fragment conjugate with a radionuclide, and recovering a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate.

23. A composition comprising an antibody according to any one of claims 1-4, an antibody fragment according to claim 5, an antibody drug conjugate according to any one of claims 6-17, a nucleic acid sequence according to claim 18, a nucleic acid construct according to claim 19, a host cell according to claim 20, an antibody or antibody fragment produced according to the method of claim 21 , or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate produced by the method according to claim 22.

24. A pharmaceutical composition comprising an antibody according to any one of claims 1-4, an antibody fragment according to claim 5, an antibody drug conjugate according to any one of claims 6-17, a nucleic acid sequence according to claim 18, a nucleic acid construct according to claim 19, a host cell according to claim 20, an antibody or antibody fragment produced according to the method of claim 21 , or a radionuclide labelled chelator-antibodyconjugate or radionuclide labelled chelator-antibody fragment conjugate produced by the method according to claim 22.

25. An antibody according to any one of claims 1-4, an antibody fragment according to claim 5, an antibody drug conjugate according to any one of claims 6-17, a nucleic acid sequence according to claim 18, a nucleic acid construct according to claim 19, a host cell according to claim 20, an antibody or antibody fragment produced according to the method of claim 21 , or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelatorantibody fragment conjugate produced by the method according to claim 22, or a composition according to any one of claims 23-24 for use as a medicament.

26. An antibody according to any one of claims 1-4, an antibody fragment according to claim 5, an antibody drug conjugate according to any one of claims 6-17, a nucleic acid sequence according to claim 18, a nucleic acid construct according to claim 19, a host cell according to claim 20, an antibody or antibody fragment produced according to the method of claim 21 , or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelatorantibody fragment conjugate produced by the method according to claim 22, or a composition according to any one of claims 23-24 for use in the treatment of cancer.

27. An antibody according to any one of claims 1-4, an antibody fragment according to claim 5, an antibody drug conjugate according to any one of claims 6-17, a nucleic acid sequence according to claim 18, a nucleic acid construct according to claim 19, a host cell according to claim 20, an antibody or antibody fragment produced according to the method of claim 21 , or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelatorantibody fragment conjugate produced by the method according to claim 22, or a composition according to any one of claims 23-24 for use in the treatment of a PTK7 expressing cancer selected from but not limited to ovarian cancer, cervical cancer, endometrial cancer, breast cancer including triple-negative breast cancer, renal cancer, bladder cancer including urothelial cancer, colorectal cancer, mesothelioma, osteosarcoma, brain cancer, including glioma, head and neck cancer, liver cancer including hepatocellular carcinoma, lung cancer including non-small-cell lung cancer, lymphoma, myeloma, leukemia, melanoma, neuroendocrine cancer including carcinoid, pancreatic cancer, , prostate cancer, testis cancer, skin cancer, gastric cancer, squamous cell carcinoma including esophageal squamous cell carcinoma, and thyroid cancer..

28. An antibody according to any one of claims 1-4, an antibody fragment according to claim 5, an antibody drug conjugate according to any one of claims 6-17, a nucleic acid sequenceaccording to claim 18, a nucleic acid construct according to claim 19, a host cell according to claim 20, an antibody or antibody fragment produced according to the method of claim 21 , or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelatorantibody fragment conjugate produced by the method according to claim 22, or a composition according to any one of claims 23-24 for use in the treatment of ovarian cancer.

29. An antibody according to any one of claims 1-4, an antibody fragment according to claim 5, an antibody drug conjugate according to any one of claims 6-17, a nucleic acid sequence according to claim 18, a nucleic acid construct according to claim 19, a host cell according to claim 20, an antibody or antibody fragment produced according to the method of claim 21 , or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelatorantibody fragment conjugate produced by the method according to claim 22, or a composition according to any one of claims 23-24 for use in the treatment of bladder cancer.

30. An antibody according to any one of claims 1-4, an antibody fragment according to claim 5, an antibody drug conjugate according to any one of claims 6-17, a nucleic acid sequence according to claim 18, a nucleic acid construct according to claim 19, a host cell according to claim 20, an antibody or antibody fragment produced according to the method of claim 21 , or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelatorantibody fragment conjugate produced by the method according to claim 22, or a composition according to any one of claims 23-24 for use in the diagnosis, monitoring or imaging of cancer.31 . An antibody according to any one of claims 1-4, an antibody fragment according to claim 5, an antibody drug conjugate according to any one of claims 6-17, a nucleic acid sequence according to claim 18, a nucleic acid construct according to claim 19, a host cell according to claim 20, an antibody or antibody fragment produced according to the method of claim 21 , or a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelatorantibody fragment conjugate produced by the method according to claim 22, or a composition according to any one of claims 23-24 for use as an imaging agent.

32. A kit for producing a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate, the kit comprisingan antibody according to any one of claims 1-4 or an antibody fragment according to claim 5, a solution of a radionuclide selected as any one of the group consisting of211At, 213Bi,212Bi,224Ra,223Ra,212Pb,225Ac,227Th,90Y,161Tb,186Re,188Re,199Au,194lr, 166Ho,159Gd,153Sm,149Pm,142Pr,111Ag,109Pd,77As,67Cu,64Cu,47Sc, and177Lu, and optionally instructions for producing the radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate using the materials provided in the kit.

33. A kit for producing a radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate, the kit comprising a chelator-antibody conjugate or complex, wherein the antibody is an antibody according to any one of claims 1-6 or an antibody fragment according to claim 7, a solution of a radionuclide selected as any one of the group consisting of211At, 213Bi,212Bi,224Ra,223Ra,212Pb,225Ac,227Th, "Y,161Tb,186Re,188Re,199Au,194lr, 166Ho,159Gd,153Sm,149Pm,142Pr,111Ag,109Pd,77As,67Cu,64Cu,47Sc, and177Lu, and optionally instructions for producing the radionuclide labelled chelator-antibody conjugate or radionuclide labelled chelator-antibody fragment conjugate using the materials provided in the kit.

34. The kit according to any one of claims 32 or 33, wherein the chelator is selected as any one of p-SCN-bn-DOTA and p-SCN-Bn-TCMC and the radionuclide is selected as any one of177Lu or212Pb.

35. The kit according to any one of claims 32-34, wherein the chelator is selected as TCMC and the radionuclide is selected as177Lu.

36. The kit according to any one of claims 32-35, wherein the chelator is selected as Bn-DOTA and the radionuclide is selected as212Pb.

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