vaccine

Abstract

The present invention relates to antigens, antibodies and vaccines for treatment or prevention of malaria.

Description

[0001] VACCINE

[0002] SEQUENCE LISTING

[0003] The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled P78923WO_Seq List for filing.xml, created on 23 July 2025, which is 168 KB in size. The information in the electronic format of the sequence listing is incorporated herein by reference in its entirety.

[0004] FIELD OF INVENTION

[0005] The present invention relates to antigens, antibodies and vaccines for treatment or prevention of malaria.

[0006] BACKGROUND

[0007] Malaria is a mosquito-borne disease caused by infection with the parasite Plasmodium. Malaria is a prevalent disease worldwide, with a particularly high prevalence in tropical and subtropical regions. The infection of red blood cells (RBCs) by the blood-stage form of the Plasmodium parasite is responsible for the clinical manifestations of malaria. Examples of Plasmodium parasite include the species P. falciparum, P. vivax, P. ovale and P. malariae. Among the most prevalent and harmful species are P. falciparum (Pf) and P. vivax (Pv).

[0008] The life cycle of Plasmodium is complex and mainly includes three stages: the pre-erythrocytic stage (or liver stage), the blood stage and the mosquito stage (or transmission stage and sexual stage). In the liver stage and blood stage, Plasmodium undergoes asexual development and reproduction in the human body, while in the mosquito stage, sexual reproduction and sporozoite proliferation and development occur in the mosquito. The life cycle of P. vivax differs in key aspects to that of P. falciparum. In particular, during the liver stage, P. vivax sporozoites may enter hepatocytes but not immediately start to grow and divide, remaining instead in a dormant, hypnozoite stage. These parasites are known as hypnozoites and may remain dormant for weeks or even months, and so can cause relapsing infections in the absence of transmission. In addition, compared with P. falciparum, P. vivax gametocytes can appear early in the parasite life cycle, before onset of clinical symptoms. P. vivax also displays a shorter development cycle in the mosquito host.

[0009] Most vaccine development to-date has focused on the major cause of malaria mortality, P. falciparum. In order to control and eliminate malaria, vaccine candidates have been designed for each of the three stages of the life cycle of P. falciparum.

[0010] Even with research prioritising P. falciparum vaccines, and the high level of clinical need, at present, only two P. falciparum vaccines have been approved by the World Health Organization (WHO) and are currently available on the market. The first is the RTS, S / AS01, developed by the GSK and the PATH organization. The second is the R21 / Matrix-M, developed by the Jenner Institute of the University of Oxford with cooperation of other organisations. The RTS, S and R21 vaccines consist of the C-terminal sequences of the circumsporozoite protein (CSP) of the P. falciparum, which have been fused with the hepatitis B virus surface antigen (HBsAg) and assembled into VLP, but they differ in the display density of the CSP antigen. However, RTS, S and R21 are prophylactic vaccines that do not protect against Plasmodium infections in the blood stage and do not block sexual reproduction and transmission in mosquitoes because their antigenic targets are P. falciparum cyclosporins (proteins expressed by Plasmodium in the pre-erythrocytic stage).

[0011] However, an effective P. vivax vaccine could also greatly facilitate elimination of P. vivax in many areas of the Americas and Asia-Pacific where standard malaria control tools are facing challenges in view of the unique aspects of P. vivax biology, such as its ability for form dormant hypnozoites and the pre-clinical appearance of gametocytes.

[0012] Currently, there is no P. vivax vaccine on the market. All the vaccine candidates in clinical trials are single stage vaccines. There are no vaccine candidates that could target multiple stages of P. vivax. A multi-stage P. vivax vaccine would be highly desirable for P. vivax malaria prevention, control, and treatment.

[0013] It is an object of the present invention is to overcome the current shortage of vaccines for P. vivax. The present invention addresses one or more of the above needs by providing P. vivax antigens and fusion proteins thereof which induce potent antibodies against P. vivax infection.

[0014] SUMMARY OF THE INVENTION

[0015] The present inventors provide for the first time a P. vivax vaccine candidate that is capable of targeting multiple stages of the P. vivax life cycle. In particular, the present inventors have developed a fusion protein comprising a P. vivax circumsporozoite protein (PvCSP) antigen and a P. vivax 230 kDa sexual-stage protein (Pvs230) antigen. Additionally, the present inventors also developed a fusion protein comprising a PvCSP antigen, a Pvs230 antigen and a Plasmodium vivax Duffy binding protein II (PVDBPII) antigen. They have shown that these fusion proteins can be expressed and are stable. Further, the fusion proteins are immunogenic, and have been demonstrated to be capable of eliciting antibodies against PvCSP, Pvs230 and PVDBPII in mice. Expression of the fusion protein on the surface of a virus-like particle (VLP) further improves stability and immunogenicity compared with the fusion protein alone.

[0016] Accordingly, the present invention provides a fusion protein comprising: (a) a Plasmodium vivax circumsporozoite protein (PvCSP) or immunogenic fragment thereof; and (b) a P. vivax 230 kDa sexual-stage protein (Pvs230) or immunogenic fragment thereof.

[0017] A fusion protein of the invention may comprise: (a) at least 10 repeat motifs derived from the central repeat region of a Type I (VK210) of P. vivax; and / or (b) at least 10 repeat motifs derived from the central repeat region of a Type II (VK247) of P. vivax. The at least 10 repeat motifs derived from the central repeat region of a Type I PvCSP may comprise at least 5 copies of the amino acid sequence GDRADGQPA (SEQ ID NO: 4) and at least 5 copies of the amino acid sequence GDRAAGQPA (SEQ ID NO: 5), wherein optionally: (i) at least one copy of SEQ ID NO: 5 is interspersed between copies of SEQ ID NO: 4; and / or (ii) the at least 10 repeat motifs derived from the central repeat region of a Type I PvCSP further comprise at least one copy of GNGAGGQAA (SEQ ID NO: 6). Alternatively or in addition, the at least 10 repeat motifs derived from the central repeat region of a Type II PvCSP may comprise at least 5 copies of the amino acid sequence ANGAGNQPG (SEQ ID NO: 7) and at least one copy of any two or more of (i) EDGAGNQPG (SEQ ID NO: 8); (ii) ANGAGDQPG (SEQ ID NO: 9); (iii) ANGADDQPG (SEQ ID NO: 10); (iv) ANGAGGQAA (SEQ ID NO: 11); and (v) GGNA ANKKAEDA GGNA GGNA (SEQ ID NO: 12).

[0018] The PvCSP protein or immunogenic fragment thereof may comprise or consist of: (a) an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 1; and / or (b) an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 2. The PvCSP protein or immunogenic fragment thereof may comprise or consist of: (a) the amino acid sequence of SEQ ID NO: 1; and / or (b) the amino acid sequence SEQ ID NO: 2.

[0019] The PvCSP protein or immunogenic fragment thereof may be a chimeric PvCSP protein or immunogenic fragment thereof which comprises (a) at least 10 repeat motifs derived from the central repeat region of a Type I PvCSP; and (b) at least 10 repeat motifs derived from the central repeat region of a Type II PvCSP; wherein optionally said PvCSP protein or immunogenic fragment is as defined herein. The chimeric PvCSP protein or immunogenic fragment thereof may comprise or consist of an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 3 or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3. The PvCSP protein or immunogenic fragment thereof may comprise or consist of an amino acid sequence of SEQ ID NO: 3.

[0020] The PvCSP protein may further comprise or consist of a PvCSP protein C-terminal region (CTR) or immunogenic fragment thereof. The PvCSP protein CTR or immunogenic fragment thereof may comprise or consist of an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 90. The PvCSP protein CTR or immunogenic fragment thereof may comprise or consist of an amino acid sequence of SEQ ID NO: 90. The PvCSP protein or immunogenic fragment thereof may comprise or consist of an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 91. The PvCSP protein or immunogenic fragment thereof may comprise or consist of the amino acid sequence of SEQ ID NO: 91.

[0021] The Pvs230 or immunogenic fragment thereof may comprise or consist of Domain 1 (DI) of PVs230, optionally with one or two substitutions. The Pvs230 or immunogenic fragment thereof may comprise or consist of an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 13 or 83, preferably SEQ ID NO: 13; or a fragment thereof which is at least 50% of the length of SEQ ID NO: 13 or 83, preferably SEQ ID NO: 13. The Pvs230 or immunogenic fragment thereof may comprise or consist of the amino acid sequence of SEQ ID NO: 13 or 83, preferably SEQ ID NO: 13.

[0022] In a fusion protein of the invention: (a) the PvCSP protein or immunogenic fragment thereof may be N-terminal to Pvs230 or the immunogenic fragment thereof; or (b) the PvCSP protein or immunogenic fragment thereof may be C-terminal to Pvs230 or the immunogenic fragment thereof; preferably the PvCSP protein or immunogenic fragment thereof is N-terminal to the Pvs230 or the immunogenic fragment thereof. A fusion protein of the invention may further comprise or consist of a Plasmodium vivax Duffy binding protein II (PVDBPII) or immunogenic fragment thereof. The DBPII or immunogenic fragment thereof may comprise or consist of an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 92. The PVDBPII or immunogenic fragment thereof may comprise or consist of the amino acid sequence of SEQ ID NO: 92, optionally comprising one, two or three substitutions. The DBPII or immunogenic fragment thereof may comprise or consist of the amino acid sequence of SEQ ID NO: 93.

[0023] In a fusion protein of the invention which comprises DBPII or immunogenic fragment thereof: (a) (i) the PVDBPII protein or immunogenic fragment thereof may be N-terminal to Pvs230 or the immunogenic fragment thereof; or (ii) the PVDBPII protein or immunogenic fragment thereof may be C-terminal to Pvs230 or the immunogenic fragment thereof; and / or (b) (i) the PVDBPII protein or immunogenic fragment thereof may be N-terminal to PvCSP or the immunogenic fragment thereof; or (ii) the PVDBPII protein or immunogenic fragment thereof may be C-terminal to PvCSP or the immunogenic fragment thereof. Preferably the PVDBPII protein or immunogenic fragment thereof may be N-terminal to Pvs230 or the immunogenic fragment thereof and C-terminal to PvCSP or the immunogenic fragment thereof.

[0024] The PvCSP protein or immunogenic fragment thereof, Pvs230 or the immunogenic fragment thereof and / or PVDBPII protein or immunogenic fragment thereof may be separated from the remainder of the fusion protein by a linker polypeptide, wherein optionally the linker polypeptide comprises the amino acid sequence of SEQ ID NO: 100.

[0025] A fusion protein of the invention may further comprise or consist of a signal peptide sequence located at the N-terminus, optionally wherein the signal peptide sequence is selected from the group consisting of: SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16.

[0026] A fusion protein of the invention may further comprise or consist of a first complexing motif optionally at the N- or C-terminus, preferably the C-terminus, and wherein optionally the first complexing motif comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 17, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37. The first complexing motif may be separated from the remainder of the fusion protein by a linker polypeptide, wherein optionally the linker polypeptide comprises the amino acid sequence of SEQ ID NO: 18, SEQ ID NO: 101, SEQ ID NO: 115, and / or SEQ ID NO: 19.

[0027] A fusion protein of the invention may further comprise or consist of a label at the N- or C-terminus, wherein optionally said label is selected from the group consisting of a histag (SEQ ID NO: 38), GST (SEQ ID NO: 39), MBP (SEQ ID NO: 40), NusA (SEQ ID NO: 41) and SUMO (SEQ ID NO: 42), preferably SEQ ID NO: 38.

[0028] A fusion protein of the invention may comprise or consist of an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to an amino acid sequence selected from the group consisting of: SEQ ID NOs: 20 to 27,63 to 66, 94 to 99, 102 to 110 and 126; preferably SEQ ID NO: 20, 21, 22, 23, 24, 94, 99 or 102 to 107; more preferably SEQ ID NO: 20, 24, 94, 99 or 102 to 107. A fusion protein of the invention may further comprise or consist of an amino acid sequence selected from the group consisting of: SEQ ID NOs: 20 to 27,63 to 66, 94 to 99, 102 to 110 and 126; preferably SEQ ID NO: 20, 22, 23, 24, 94, 99 or 102 to 107; more preferably SEQ ID NO: 20, 24, 94, 99 or 102 to 107.

[0029] The invention further provides a chimeric PvCSP protein or immunogenic fragment thereof, which comprises (a) at least 10 repeat motifs derived from the central repeat region of a Type I PvCSP; and (b) at least 10 repeat motifs derived from the central repeat region of a Type II PvCSP; wherein said PvCSP protein or immunogenic fragment is as defined herein, and preferably said PvCSP protein or immunogenic fragment comprises an amino acid sequence of SEQ ID NO: 3, 56 to 62 or 84.

[0030] The invention also provides a chimeric PvCSP protein or immunogenic fragment thereof, which comprises (a) at least 10 repeat motifs derived from the central repeat region of a Type I PvCSP; (b) at least 10 repeat motifs derived from the central repeat region of a Type II PvCSP; and (c) a PvCSP protein C -terminal region (CTR) or immunogenic fragment thereof; wherein said PvCSP protein or immunogenic fragment is as defined herein, and preferably said PvCSP protein or immunogenic fragment comprises an amino acid sequence of SEQ ID NO: 91 or 111-114.

[0031] The invention also provides a virus-like particle (VLP) comprising: (a) at least one fusion protein of the invention; (b) at least one chimeric PvCSP protein or immunogenic fragment thereof of the invention; and / or (c) at least one Pvs230 or immunogenic fragment thereof of the invention; and / or at least one PVDBPII protein or immunogenic fragment thereof of the invention.

[0032] A VLP of the invention may comprise a core which comprises at least one subunit of a VLP carrier, optionally the at least one subunit of the VLP carrier is selected from the group consisting of: an amino acid sequence of SEQ ID NO: 28, Hepatitis B surface antigen (SEQ ID NO: 43), HPV 18 LI protein (SEQ ID NO: 44), Hepatitis E P239 protein (SEQ ID NO: 45), HPV 16 LI protein (SEQ ID NO: 46), AP205 (SEQ ID NO: 47), ferritin (SEQ ID NO: 48), I53-50A (SEQ ID NO: 49) or I53-50B (SEQ ID NO: 69), preferably an amino acid sequence of SEQ ID NO: 28. Alternatively or in addition, in a VLP of the invention, the at least one fusion protein, at least one chimeric PvCSP protein or immunogenic fragment thereof and / or at least one Pvs230 or immunogenic fragment thereof is covalently linked to the at least one subunit of the VLP carrier, such that the at least one fusion protein, at least one chimeric PvCSP protein or immunogenic fragment thereof and / or at least one Pvs230 or immunogenic fragment thereof is presented on the surface of the VLP.

[0033] The at least one subunit of the VLP carrier may further comprise or consist of a second complexing motif at the N- or C-terminus of the at least one subunit of the VLP carrier, preferably at the N-terminus of the at least one subunit of the VLP carrier; wherein optionally the second complexing motif comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 29, pilin-N (SEQ ID NO: 50), pilin-C (SEQ ID NO: 51), SnoopCatcher (SEQ ID NO: 52), SdyCatcher (SEQ ID NO: 53), SilkCatcher (SEQ ID NO: 54) or DogCatcher (SEQ ID NO: 55), preferably SEQ ID NO: 29. The second complexing motif may be separated from the remainder of the at least one subunit of the VLP carrier by a linker polypeptide, wherein optionally the linker polypeptide comprises the amino acid sequence of SEQ ID NO: 18 and / or SEQ ID NO: 19.

[0034] The VLP core may comprise or consist of an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent. The VLP core may comprise or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent.

[0035] In a VLP of the invention, the first complexing motif and the second complexing motif may be covalently linked, such that the at least one fusion protein, at least one chimeric PvCSP protein or immunogenic fragment thereof and / or at least one Pvs230 or immunogenic fragment thereof; and / or at least one PVDBPII protein or immunogenic fragment thereof, is presented on the surface of the VLP.

[0036] A VLP of the invention may comprise or consist of: (a) a fusion protein comprising or consisting of an amino acid sequence selected from the group consisting of: SEQ ID NOs: 20 to 27,63 to 66, 94 to 99, 102 to 110 and 126, preferably any one of SEQ ID NOs: 20 to 24, 94, 99 or 102 to 107; more preferably SEQ ID NO: 20, 24, 94, 99 or 102 to 107; and (b) a VLP core comprising or consisting of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent.

[0037] A VLP of the invention may have a diameter of less than about 100 nm or less than about 75 nm, preferably the VLP has a diameter of between about 30 nm to about 55 nm, optionally as measured by dynamic light scattering (DLS) and / or electron microscopy.

[0038] The invention further provides a nucleic acid encoding a fusion protein of the invention, a chimeric PvCSP protein or immunogenic fragment thereof of the invention, a Pvs230 or immunogenic fragment thereof of the invention, or a VLP of the invention, wherein optionally said nucleic acid: (a) comprises or consists of a nucleic acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 31, 74-82, or 116 to 125, preferably SEQ ID NO: 31, 120 or 125; or (b) comprises or consists of the nucleic acid sequence of SEQ ID NO: 31, 74-82 or 116 to 125, preferably SEQ ID NO: 31, 120 or 125. The invention also provides a viral vector, RNA vector or DNA plasmid comprising a nucleotide sequence of the invention, optionally operably linked to a promoter, wherein optionally the viral vector is an adenoviral vector, a vesicular stomatitis virus (VSV) vector, an adeno-associated virus (AAV) vectors, a measles virus vector, a retroviral vector, a Herpesvirus vector, an influenza virus vector or a modified Vaccinia Ankara viral vector, preferably a non-replicating adenoviral vector or a non-replicating modified Vaccinia Ankara viral vector.

[0039] The invention further provides a host cell comprising the nucleotide sequence or DNA plasmid of the invention, wherein optionally the host cell is a mammalian cell, a yeast cell or a bacterial cell.

[0040] The invention also provides a nucleic acid, viral vector, RNA vector or DNA plasmid or host cell of the invention, which further comprises or consists of a nucleic acid sequence encoding a VLP core, and preferably encoding a VLP core with the amino acid sequence of SEQ. ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent.

[0041] The invention further provides a pharmaceutical composition comprising a fusion protein, a chimeric PvCSP protein or immunogenic fragment thereof, a Pvs230 or immunogenic fragment thereof, a VLP, a nucleic acid, or a viral vector or DNA plasmid of the invention, and optionally one or more pharmaceutically acceptable excipient, carrier and / or diluent.

[0042] The invention also provides a vaccine composition comprising a fusion protein, a chimeric PvCSP protein or immunogenic fragment thereof, a Pvs230 or immunogenic fragment thereof, a VLP, a nucleic acid, or a viral vector or DNA plasmid of the invention, and a pharmaceutically acceptable adjuvant.

[0043] In a vaccine composition of the invention, the pharmaceutically acceptable adjuvant may be selected from MF59, AS01 or biosimilars thereof, or a combination thereof.

[0044] A pharmaceutical composition or a vaccine composition of the invention may be stable at 4°C, 25°C and 37°C for at least 2 weeks. A pharmaceutical composition or a vaccine composition of the invention may be stable at -80°C for at least 3 months.

[0045] A pharmaceutical composition or a vaccine composition of the invention may be a liquid or solid formulation.

[0046] A vaccine composition of the invention may induce antibodies with a geometric mean titre (GMT) of: (a) at least 50,000 IU / mL, optionally at least 60,000 IU / mL against PvCSP; and / or (b) at least 5,000 IU / mL, optionally at least 6,000 IU / mL, at least 7,000 IU / mLor at least 8,000 IU / mL against Pvs230; and / or (c) at least 50,000 IU / mL, optionally at least 60,000 IU / mL against PVDBPII.

[0047] The invention further provides a pharmaceutical preparation comprising: (a) a pharmaceutical composition or a vaccine composition of the invention; and (b) (i) one or more additional P. vivax polypeptide, or a variant, derivative or fragment thereof, or a nucleic acid encoding said polypeptide; and / or (ii) one or more P. falciparum polypeptide, or a variant, derivative or fragment thereof, or a nucleic acid encoding said polypeptide; as a combined preparation in a form suitable for simultaneous, separate or sequential use. The invention also provides a method of producing a fusion protein, a chimeric PvCSP protein or immunogenic fragment thereof, a Pvs230 or immunogenic fragment thereof, or a VLP of the invention, the method comprising culturing a host cell of the invention under conditions suitable to express said fusion protein, chimeric PvCSP protein or immunogenic fragment thereof, Pvs230 or immunogenic fragment thereof, or VLP, and recovering the fusion protein, chimeric PvCSP protein or immunogenic fragment thereof, Pvs230 or immunogenic fragment thereof, or VLP produced, optionally wherein said method further comprises: (a) introducing a nucleic acid of the invention into said host cell; and / or (b) purifying the fusion protein, chimeric PvCSP protein or immunogenic fragment thereof, Pvs230 or immunogenic fragment thereof, or VLP produced.

[0048] The invention further provides a fusion protein, a chimeric PvCSP protein or immunogenic fragment thereof, a Pvs230 or immunogenic fragment thereof, a VLP, a nucleic acid, a viral vector, RNA vector or DNA plasmid, a pharmaceutical composition, a vaccine composition, or a pharmaceutical preparation of the invention for use in a method of preventing and / or treating malaria.

[0049] The invention also provides a method of preventing and / or treating malaria comprising the administration of an effective dose of fusion protein, a chimeric PvCSP protein or immunogenic fragment thereof, a Pvs230 or immunogenic fragment thereof, a VLP, a nucleic acid, a viral vector, RNA vector or DNA plasmid, a pharmaceutical composition, a vaccine composition, or a pharmaceutical preparation of the invention to a patient in need thereof.

[0050] The invention further provides the use of a fusion protein, a chimeric PvCSP protein or immunogenic fragment thereof, a Pvs230 or immunogenic fragment thereof, a VLP, a nucleic acid, or a viral vector or DNA plasmid of the invention, in the manufacture of a medicament for the prevention and / or treatment of malaria.

[0051] Brief Description of the Figures

[0052] Figure 1 - Optimisation of PV2 target fusion protein expression in CHO cells. (A) SDS PAGE showing PV2 expression level at day 3 (D3) to day 10 (DIO). (B) Western Blot showing PV2 expression level at day 3 (D3) to day 10 (DIO) using anti-Histag primary antibody.

[0053] Figure 2 - Nickel Affinity Chromatography purification of PV2 target fusion protein. (A) Chromatography trace showing elution fractions for the purification of PV2 target fusion protein. (B) SDS PAGE showing fractions A4, A6, A8, A10, A12, B2, B4 and B6 as well as outlet (flow-through), wash and supernatant (input). Figure 3 - Size Exclusion Chromatography (SEC) purification of PV2 target fusion protein. (A) Chromatography trace showing elution fractions for the purification of PV2 target fusion protein. (B) SDS PAGE showing fractions E9, E11, F1, F3, F5, F7, F11 and G7.

[0054] Figure 4 - Size Exclusion Chromatography purification of PV2-VLP. (A) Chromatography trace showing elution fractions for the purification of PV2-VLP. (B) SDS PAGE showing input, PV2-VLP and PV2.

[0055] Figure 5 - PV2 purity characterisation by SEC-HPLC showing high purity of the PV2 target fusion protein after Nickel Affinity Chromatography purification and SEC purification.

[0056] Figure 6 - Dynamic Light Scattering analysis of PV2-VLP showing uniform size of the particles.

[0057] Figure 7 - PV2-VLP purity characterisation by SEC-HPLC showing generally high purity of over 73.57% of the area under the curve and a residual polymer peak accounting for 26.43% of the area under the curve.

[0058] Figure 8 - Western Blot showing PV2-VLP display efficiency compared with VLP alone. The display efficiencies were 61.4% and 68.3% respectively.

[0059] Figure 9 - Electron Microscopy imaging of particles: (A) PV2-VLP. Scale bar is 100 nm. (B) PV5-VLP. Scale bar is 100 nm. (C) PV16-VLP. Scale bar is 200 nm. (D) PV33-VLP. Scale bar is 200 nm.

[0060] Figure 10 - PV2-VLP stability. (A) SDS PAGE showing stability of PV2-VLP over time at 4°C, 25°C and 37°C and shown at day 1, day 2, day 3, day 7 and day 14 showing stability of PV2-VLP. (B) DLS detection of particle size over time at 4°C, 25°C and 37°C showing relatively stable particle size of PV2-VLP with moderate aggregation at day 14 for 25°C and 37°C (measured in nm).

[0061] Figure 11 - PV2-VLP long term stability. (A) SDS PAGE showing stability of PV2-VLP over time at -80°C and shown at day 0, 1 month, 2 months and 3 months showing stability of PV2-VLP. (B) DLS detection of particle size over time at -80°C showing stable particle size of PV2-VLP (measured in nm).

[0062] Figure 12 - Immunisation experiment using PV2, PV5, PV2-VLP (PV2M) and PV5-VLP (PV5M). (A) and (B) Experimental plan. (C) ELISA results using anti-PvCSP and anti-Pvs230 primary antibodies. PV2, PV5, PV2-VLP and PV5-VLP elicit a strong immune response in mice. Differences are not statistically significant between groups immunised with PV2 and PV2-VLP at either day 20 or day 42. Graphs are displayed in logarithmic scale. Figure 13 - Adjuvant optimisation for PV2-VLP immunisation in vivo. (A) and (B) Experimental plan. (C) ELISA results at day 20 using anti-PvCSP and anti-Pvs230 primary antibodies. (D) ELISA results at day 42 using anti-PvCSP and anti-Pvs230 primary antibodies. Graphs are displayed in logarithmic scale.

[0063] Figure 14 -Cellular immune response of PV2-VLP immunisation. (A) and (B) Experimental plan. (C) ELISpot results showing expression levels of IFN-γ at day 42. (D) ELISpot results showing expression levels of IL-2 at day 42. Graphs are displayed in logarithmic scale.

[0064] Figure 15 - Expression in CHO cells of PV1, PV2, PV4, PV5, PV6, PV16, PV17, PV18, PV19, PV20, PV33 target fusion protein. (A) SDS PAGE of the supernatant in the expression stage of CHO cells of PV1, PV2, PV4, PV5 and PV6. (B) SDS PAGE of the supernatant in the expression stage of CHO cells of PV16, PV17, PV18, PV19 and PV20. (C) SDS PAGE of the supernatant in the expression stage of CHO cells of PV33. (D) Western Blot showing PV16, PV17, PV18, PV19 and PV20 using anti-His primary antibody. (E) Western Blot showing PV33 using anti-His primary antibody.

[0065] Figure 16 - Size Exclusion Chromatography (SEC) purification of PV5 target fusion protein. (A) Chromatography trace showing elution fractions for the purification of PV5 target fusion protein. (B) SDS PAGE showing fractions B5, B12, Cl, C2, C3, C4, C5, C6 and C7.

[0066] Figure 17 - Size Exclusion Chromatography (SEC) purification of PV16 target fusion protein. (A) Chromatography trace showing elution fractions for the purification of PV16 target fusion protein. (B) SDS PAGE showing fractions B3, B5, B7, B11, C1, C3, C5 and C7.

[0067] Figure 18 - Size Exclusion Chromatography (SEC) purification of PV17 target fusion protein. (A) Chromatography trace showing elution fractions for the purification of PV17 target fusion protein. (B) SDS PAGE showing fractions A9, A11, B1, B3, B5, B7, B9, B11, C2 and C4.

[0068] Figure 19 - Size Exclusion Chromatography (SEC) purification of PV18 target fusion protein. (A) Chromatography trace showing elution fractions for the purification of PV18 target fusion protein. (B) SDS PAGE showing fractions H8, H10, A2, A4, A6, A8, AID, A12 and B2.

[0069] Figure 20 - Size Exclusion Chromatography (SEC) purification of PV19 target fusion protein. (A) Chromatography trace showing elution fractions for the purification of PV19 target fusion protein. (B) SDS PAGE showing fractions E4, E6, E8, E10, E12, F2, F4, F6, F8, F10 and F12. Figure 21 - Size Exclusion Chromatography (SEC) purification of PV20 target fusion protein. (A) Chromatography trace showing elution fractions for the purification of PV20 target fusion protein. (B) SDS PAGE showing fractions E12, F8, F10, F12, G2, G4, G6, G8, GIO, G12 and H2.

[0070] Figure 22 - Size Exclusion Chromatography (SEC) purification of PV33 target fusion protein. (A) Chromatography trace showing elution fractions for the purification of PV33 target fusion protein. (B) SDS PAGE showing fractions D2, D4, D6, D8 and Dll.

[0071] Figure 23 - SEC purification of PV5-VLP. (A) Chromatography trace showing elution fractions for the purification of PV5-VLP. (B) SDS PAGE showing fractions B4, B5 and B6.

[0072] Figure 24 - SEC purification of PV16-VLP. (A) Chromatography trace showing elution fractions for the purification of PV16-VLP. (B) SDS PAGE showing fractions C3, C4 and C5.

[0073] Figure 25 - SEC purification of PV17-VLP. (A) Chromatography trace showing elution fractions for the purification of PV17-VLP. (B) SDS PAGE showing fractions H2, H3 and H5.

[0074] Figure 26 - SEC purification of PV18-VLP. (A) Chromatography trace showing elution fractions for the purification of PV18-VLP. (B) SDS PAGE showing fractions B3, B4, B5, B6, B7, B8 and C4.

[0075] Figure 27 - SEC purification of PV19-VLP. (A) Chromatography trace showing elution fractions for the purification of PV19-VLP. (B) SDS PAGE showing fractions C3, C4, C5, C6 and C7.

[0076] Figure 28 - SEC purification of PV20-VLP. (A) Chromatography trace showing elution fractions for the purification of PV20-VLP. (B) SDS PAGE showing fractions F4, F5, F6, F7, F8 and G5.

[0077] Figure 29 - SEC purification of PV33-VLP. (A) Chromatography trace showing elution fractions for the purification of PV20-VLP. (B) SDS PAGE showing PV33 reduced (lane 3), PV33 non-reduced (lane 4), PV33-VLP reduced (lane 5), PV33-VLP non-reduced (lane 6).

[0078] Figure 30 - Dynamic Light Scattering analysis of (A) PV16-VLP and (B) PV33-VLP showing uniform size of the particles.

[0079] Figure 31 - Immunisation experiment using PV2, PV16, PV33 fusion proteins and PV2-VLP, PV16-VLP and PV33-VLP (PV2M, PV16M and PV33M). (A) and (B) Experimental plan. (C) ELISA results using anti-PvCSP primary antibodies on Day l sample. (D) ELISA results using anti-PvDBPII primary antibodies on Day 27 sample. (E) ELISA results using anti-Pvs230 primary antibodies on Day 1 sample. PV33 elicited higher antibody titres against PvCSP and PvS230 compared to PV2 or PV16. Nanoparticle PV33-VLP demonstrated comparable immunogenicity to PV16-VLP and PV2-VLP in anti-PvCSP and Pvs230 tiers. Nanoparticle PV33-VLP demonstrated comparable immunogenicity to PV16-VLP in anti-PvDBPIL Graphs are displayed in logarithmic scale.

[0080] DETAILED DESCRIPTION

[0081] Definitions

[0082] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Singleton, et al., DICTIONARY OF MICROBIOLOGY AND MOLECULAR BIOLOGY, 20 ED., John Wiley and Sons, New York (1994), and Hale & Marham, THE HARPER COLLINS DICTIONARY OF BIOLOGY, Harper Perennial, NY (1991) provide the skilled person with a general dictionary of many of the terms used in this disclosure. The meaning and scope of the terms should be clear; however, in the event of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. It should be understood that this invention is not limited to the particular methodology, protocols, and reagents, etc., described herein and as such can vary.

[0083] The description of embodiments of the disclosure is not intended to be exhaustive or to limit the disclosure to the precise form disclosed. While specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as those skilled in the relevant art will recognize. For example, while method steps or functions are presented in a given order, alternative embodiments may perform functions in a different order, or functions may be performed substantially concurrently. The teachings of the disclosure provided herein can be applied to other procedures or methods as appropriate. The various embodiments described herein can be combined to provide further embodiments. Aspects of the disclosure can be modified, if necessary, to employ the compositions, functions and concepts described herein to provide yet further embodiments of the disclosure. Moreover, due to biological functional equivalency considerations, some changes can be made in protein structure without affecting the biological or chemical action in kind or amount. These and other changes can be made to the disclosure in light of the detailed description. All such modifications are intended to be included within the scope of the appended claims.

[0084] The headings provided herein are not limitations of the various aspects or embodiments of this disclosure.

[0085] As used herein, the term "capable of' when used with a verb, encompasses or means the action of the corresponding verb. For example, "capable of interacting" also means interacting, "capable of inducing" also means induces, "capable of binding" also means binds and "capable of specifically targeting..." also means specifically targets.

[0086] Numeric ranges are inclusive of the numbers defining the range. Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limits of that range is also specifically disclosed. Each smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in that stated range is encompassed within this disclosure. The upper and lower limits of these smaller ranges may independently be included or excluded in the range, and each range where either, neither or both limits are included in the smaller ranges is also encompassed within this disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in this disclosure.

[0087] As used herein, the articles "a" and "an" may refer to one or to more than one (e.g. to at least one) of the grammatical object of the article. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. In this application, the use of "or" means "and / or" unless stated otherwise. Furthermore, the use of the term "including", as well as other forms, such as "includes" and "included", is not limiting.

[0088] " About" may generally mean an acceptable degree of error for the quantity measured given the nature or precision of the measurements. Exemplary degrees of error are within 20 percent (%), typically, within 10%, and more typically, within 5% of a given value or range of values. Preferably, the term "about" shall be understood herein as plus or minus (±) 5%, preferably ± 4%, ± 3%, ± 2%, ± 1%, ± 0.5% or ± 0.1%, of the numerical value of the number with which it is being used.

[0089] The term "consisting of" refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not recited in that description of the invention.

[0090] As used herein the term "consisting essentially of" refers to those elements required for a given invention. The term permits the presence of elements that do not materially affect the basic and novel or functional characteristic(s) of that invention (i.e. inactive or non-immunogenic ingredients).

[0091] Embodiments described herein as "comprising" one or more features may also be considered as disclosure of the corresponding embodiments "consisting of" and / or "consisting essentially of" such features.

[0092] Concentrations, amounts, volumes, percentages and other numerical values may be presented herein in a range format. It is also to be understood that such range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. A "vector" or "construct" (sometimes referred to as gene delivery or gene transfer "vehicle") refers to a macromolecule or complex of molecules comprising a polynucleotide to be delivered to a host cell, either in vitro or in vivo. A vector can be a linear or a circular molecule. A vector of the invention may be viral or non-viral. All disclosure herein in relation vectors of the invention applies equally to viral and non-viral vectors unless otherwise stated. All disclosure in relation to viral vectors of the invention applies equally and without reservation to adenovirus vectors or pox virus vectors.

[0093] As used herein, the term "plasmid", refers to a common type of non-viral vector. A plasmid is an extra-chromosomal DNA molecule separate from the chromosomal DNA which is capable of replicating independently of the chromosomal DNA. Preferably a plasmid is circular and may be double-stranded.

[0094] The terms "nucleic acid cassette", "nucleic acid construct", "expression cassette" and "nucleic acid expression cassette" are used interchangeably to mean a nucleic acid moleculethat is capable of directing transcription. A nucleic acid cassette includes, at the least, a promoter or a structure functionally equivalent to a promoter and a nucleic acid sequence to be transcribed. Thus, a nucleic acid cassette includes, at the least, a promoter or a structure functionally equivalent to a promoter and a nucleic acid sequence encoding a protein of interest. In the present invention, a nucleic acid cassette includes, at the least, a promoter or a structure functionally equivalent to a promoter, and a nucleic acid encoding a therapeutic protein. A nucleic acid cassette may include additional elements, such as an enhancer, and / or a transcription termination signal.

[0095] As used herein, the term "virus-like particle" (VLP) refers to a particle which resembles a virus but which does not contain viral nucleic acid and is therefore non-infectious. VLPs commonly contain one or more virus capsid or envelope proteins which are capable of self-assembly to form the VLP. VLPs have been produced from components of a wide variety of virus families (Noad and Roy (2003), Trends in Microbiology, 11:438-444; Grgacic et al., (2006), Methods, 40:60-65). Some VLPs have been approved as prophylactic vaccines, for example Engerix-B (for hepatitis B), Cervarix and Gardasil (for human papilloma viruses).

[0096] As used herein, the terms "transduced" and "modified" are used interchangeably to describe cells which have been modified to express a transgene of interest, particularly an antigen as defined herein. Typically the modification occurs through transduction of the cells.

[0097] As used herein, the terms "titre" and "yield" are used interchangeably to mean the amount of viral vector produced by a method of the invention.

[0098] Amino acids are referred to herein using the name of the amino acid, the three-letter abbreviation or the single letter abbreviation.

[0099] Unless otherwise indicated, any nucleic acid sequences are written left to right in 5' to 3' orientation; amino acid sequences are written left to right in amino to carboxy orientation, respectively.

[0100] As used herein, the terms "protein" and "polypeptide" are used interchangeably herein to designate a series of amino acid residues, connected to each other by peptide bonds between the alpha- amino and carboxyl groups of adjacent residues. The terms "protein", and "polypeptide" refer to a polymer of amino acids, including modified amino acids (e.g., phosphorylated, glycated, glycosylated, etc.) and amino acid analogues, regardless of its size or function. " Protein" and "polypeptide" are often used in reference to relatively large polypeptides, whereas the term "peptide" is often used in reference to small polypeptides, but usage of these terms in the art overlaps. The terms "protein" and "polypeptide" are used interchangeably herein when referring to a gene product and fragments thereof. Thus, exemplary polypeptides or proteins include gene products, naturally occurring proteins, homologs, orthologs, paralogs, fragments and other equivalents, variants, fragments, and analogues of the foregoing.

[0101] As used herein, the terms "polynucleotides", "nucleic acid" and "nucleic acid sequence" refers to any molecule, preferably a polymeric molecule, incorporating units of ribonucleic acid, deoxyribonucleic acid or an analogue thereof. The nucleic acid can be either single-stranded or double-stranded. A singlestranded nucleic acid can be one nucleic acid strand of a denatured double- stranded DNA Alternatively, it can be a single-stranded nucleic acid not derived from any double-stranded DNA. In one aspect, the nucleic acid can be DNA. In another aspect, the nucleic acid can be RNA Suitable nucleic acid molecules are DNA, including genomic DNA or cDNA. Other suitable nucleic acid molecules are RNA, including siRNA, shRNA, and antisense oligonucleotides. The terms "transgene" and "gene" are also used interchangeably and both terms encompass fragments or variants thereof encoding the target protein, specifically an antigen of the invention.

[0102] The transgenes of the present invention include nucleic acid sequences that have been removed from their naturally occurring environment, recombinant or cloned DNA isolates, and chemically synthesized analogues or analogues biologically synthesized by heterologous systems.

[0103] Minor variations in the amino acid sequences of the invention are contemplated as being encompassed by the present invention, providing that the variations in the amino acid sequence(s) maintain at least 60%, at least 70%, more preferably at least 80%, at least 85%, at least 90%, at least 95%, and most preferably at least 97% or at least 99% sequence identity to the amino acid sequence of the invention or a fragment thereof as defined anywhere herein. The term homology is used herein to mean identity. As such, the sequence of a variant or analogue sequence of an amino acid sequence of the invention may differ on the basis of substitution (typically conservative substitution) deletion or insertion. Proteins comprising such variations are referred to herein as variants.

[0104] Proteins of the invention may include variants in which amino acid residues from one species are substituted for the corresponding residue in another species, either at the conserved or non-conserved positions. Variants of protein molecules disclosed herein may be produced and used in the present invention. Following the lead of computational chemistry in applying multivariate data analysis techniques to the structure / property-activity relationships [see for example, Wold, et al. Multivariate data analysis in chemistry. Chemometrics-Mathematics and Statistics in Chemistry (Ed.: B. Kowalski); D. Reidel Publishing Company, Dordrecht, Holland, 1984 (ISBN 90-277-1846-6] quantitative activity-property relationships of proteins can be derived using well-known mathematical techniques, such as statistical regression, pattern recognition and classification [see for example Norman et al. Applied Regression Analysis. Wiley-Interscience; 3rd edition (April 1998) ISBN: 0471170828; Kandel, Abraham et al. Computer-Assisted Reasoning in Cluster Analysis. Prentice Hall PTR, (May 11, 1995), ISBN: 0133418847; Krzanowski, Wojtek. Principles of Multivariate Analysis: A User's Perspective (Oxford Statistical Science Series, No 22 (Paper)). Oxford University Press; (December 2000), ISBN: 0198507089; Witten, Ian H. et al Data Mining: Practical Machine Learning Tools and Techniques with Java Implementations. Morgan Kaufmann; (October 11, 1999), ISBN:1558605525; Denison David G. T. (Editor) et al Bayesian Methods for Nonlinear Classification and Regression (Wiley Series in Probability and Statistics). John Wiley & Sons; (July 2002), ISBN: 0471490369; Ghose, Arup K. et al. Combinatorial Library Design and Evaluation Principles, Software, Tools, and Applications in Drug Discovery. ISBN: 0-8247-0487-8]. The properties of proteins can be derived from empirical and theoretical models (for example, analysis of likely contact residues or calculated physicochemical property) of proteins sequence, functional and three-dimensional structures and these properties can be considered individually and in combination.

[0105] Amino acids are referred to herein using the name of the amino acid, the three-letter abbreviation or the single letter abbreviation. The term "protein", as used herein, includes proteins, polypeptides, and peptides. As used herein, the term "amino acid sequence" is synonymous with the term "polypeptide" and / or the term "protein". In some instances, the term "amino acid sequence" is synonymous with the term "peptide". The terms "protein" and "polypeptide" are used interchangeably herein. In the present disclosure and claims, the conventional one-letter and three-letter codes for amino acid residues may be used. The 3-letter code for amino acids as defined in conformity with the IUPACIUB Joint Commission on Biochemical Nomenclature (JCBN). It is also understood that a polypeptide may be coded for by more than one nucleotide sequence due to the degeneracy of the genetic code.

[0106] Amino acid residues at non-conserved positions may be substituted with conservative or nonconservative residues. In particular, conservative amino acid replacements are contemplated.

[0107] A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (e.g., lysine, arginine, or histidine), acidic side chains (e.g., aspartic acid or glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, or cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, or tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, or histidine). Thus, if an amino acid in a polypeptide is replaced with another amino acid from the same side chain family, the amino acid substitution is considered to be conservative. The inclusion of conservatively modified variants in a protein of the invention does not exclude other forms of variant, for example polymorphic variants, interspecies homologs, and alleles. " Non-conservative amino acid substitutions" include those in which (i) a residue having an electropositive side chain (e.g., Arg, His or Lys) is substituted for, or by, an electronegative residue (e.g., Glu or Asp), (ii) a hydrophilic residue (e.g., Ser or Thr) is substituted for, or by, a hydrophobic residue (e.g., Ala, Leu, He, Phe or Vai), (iii) a cysteine or proline is substituted for, or by, any other residue, or (iv) a residue having a bulky hydrophobic or aromatic side chain (e.g., Vai, His, He or Trp) is substituted for, or by, one having a smaller side chain (e.g., Ala or Ser) or no side chain (e.g., Gly).

[0108] " Insertions" or "deletions" are typically in the range of about 1, 2, or 3 amino acids. The variation allowed may be experimentally determined by systematically introducing insertions or deletions of amino acids in a protein using recombinant DNA techniques and assaying the resulting recombinant variants for activity. This does not require more than routine experiments for a skilled person.

[0109] A "fragment" of a polypeptide comprises at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 97% or more of the original polypeptide. For example, a fragment may comprise at least 5, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, at least 100, at least 150, at least 200, at least 250, at least 300, at least 350, at least 400 or more amino acids of the protein from which it is derived. Preferably a fragment may comprise no more than 50, no more than 60, no more than 70, no more than 80, no more than 90, no more than 100, no more than 150, no more than 200, no more than 250 amino acids of the protein from which it is derived. A fragment may be continuous or discontinuous, preferably continuous.

[0110] A polypeptide may be "chimeric". A chimeric polypeptide may comprise amino acid sequences from two or more (e.g. from two, three, four, five or more) different proteins. These proteins may be different proteins, or may be variants of the same protein, such as variants of the protein from different strains of P. vivax.

[0111] A PvCSP antigen of the invention may be chimeric. A chimeric PvCSP antigen of the invention may comprise amino acid sequences from two or more (e.g. from two, three, four, five or more) different PvCSP antigens. Said two or more different PvCSP antigens may be from different strains of P. vivax. By way of non-limiting example, a chimeric PvCSP antigen may comprise one or more amino acid sequence from the CSP protein of the P. vivax VK210 genotype and one or more amino acid sequence from the CSP protein of the P. vivax VK247 genotype.

[0112] A Pvs230 antigen of the invention may be chimeric. A chimeric Pvs230 antigen of the invention may comprise amino acid sequences from two or more (e.g. from two, three, four, five or more) different Pvs230 antigens. Said two or more different Pvs230 antigens may be from different strains of P. vivax. By way of non-limiting example, a chimeric Pvs230 antigen may comprise one or more amino acid sequence from the Pvs230 protein of the P. vivax VK210 strain and one or more amino acid sequence from the Pvs230 protein of the P. vivax VK247 strain. A PVDBPII antigen of the invention may be chimeric. A chimeric PVDBPII antigen of the invention may comprise amino acid sequences from two or more (e.g. from two, three, four, five or more) different PVDBPII antigens. Said two or more different PVDBPII antigens may be from different strains of P. vivax. By way of non-limiting example, a chimeric PVDBPII antigen may comprise one or more amino acid sequence from the PVDBPII protein of the P. vivax VK210 strain and one or more amino acid sequence from the PVDBPII protein of the P. vivax VK247 strain.

[0113] As used herein, the term "repeat motif" refers to a short amino acid sequence, typically 5 to 15 amino acids in length, that is repeated within the central repeat region of PvCSP. The central repeat region of native PvCSP (either the VK210 or VK247 strains) may comprise up to 25 repeat motifs, and typically contains 20 repeat motifs.

[0114] As used herein, the term "copy" in the context of a repeat motif refers to the number of instances of that repeat motif present in a polypeptide, e.g. a PvCSP protein or immunogenic fragment thereof according to the invention. For example, if a polypeptide contains two copies of a repeat motif, there are two instances of said repeat motif within the polypeptide; if a polypeptide contains six copies of a repeat motif, there are six instances of said repeat motif within the polypeptide.

[0115] The polynucleotides of the present invention may be prepared by any means known in the art. For example, large amounts of the polynucleotides may be produced by replication in a suitable host cell. The natural or synthetic DNA fragments coding for a desired fragment will be incorporated into recombinant nucleic acid constructs, typically DNA constructs, capable of introduction into and replication in a prokaryotic or eukaryotic cell. Usually the DNA constructs will be suitable for autonomous replication in a unicellular host, such as yeast or bacteria, but may also be intended for introduction to and integration within the genome of a cultured insect, mammalian, plant or other eukaryotic cell lines.

[0116] The polynucleotides of the present invention may also be produced by chemical synthesis, e.g. by the phosphoramidite method or the tri-ester method and may be performed on commercial automated oligonucleotide synthesizers. A double-stranded fragment may be obtained from the single stranded product of chemical synthesis either by synthesizing the complementary strand and annealing the strand together under appropriate conditions or by adding the complementary strand using DNA polymerase with an appropriate primer sequence.

[0117] When applied to a nucleic acid sequence, the term "isolated" in the context of the present invention denotes that the polynucleotide sequence has been removed from its natural genetic milieu and is thus free of other extraneous or unwanted coding sequences (but may include naturally occurring 5' and 3' untranslated regions such as promoters and terminators) and is in a form suitable for use within genetically engineered protein production systems. Such isolated molecules are those that are separated from their natural environment. In view of the degeneracy of the genetic code, considerable sequence variation is possible among the polynucleotides of the present invention. Degenerate codons encompassing all possible codons for a given amino acid are set forth below:

[0118] Amino Acid Codons Degenerate Codon

[0119] Cys TGC TGT TGY

[0120] Ser AGC AGT TCA TCC TCG TCT WSN

[0121] Thr ACA ACC ACG ACT ACN

[0122] Pro CCA CCC CCG CCT CCN

[0123] Ala GCA GCC GCG GCT GCN

[0124] Gly GGA GGC GGG GGT GGN

[0125] Asn AAC AAT AAY

[0126] Asp GAC GAT GAY

[0127] Glu GAA GAG GAR

[0128] Gin CAA CAG CAR

[0129] His CAC CAT CAY

[0130] Arg AGA AGG CGA CGC CGG CGT MGN

[0131] Lys AAA AAG AAR

[0132] Met ATG ATG

[0133] lie ATA ATC ATT ATH

[0134] Leu CTA CTC CTG CTT TTA TTG YTN

[0135] Vai GTA GTC GTG GTT GTN

[0136] Phe TTCTTT TTY

[0137] Tyr TAC TAT TAY

[0138] Trp TGG TGG

[0139] Ter TAA TAG TGA TRR

[0140] Asn / Asp RAY

[0141] Glu / Gin SAR

[0142] Any NNN

[0143]

[0144] One of ordinary skill in the art will appreciate that flexibility exists when determining a degenerate codon, representative of all possible codons encoding each amino acid. For example, some polynucleotides encompassed by the degenerate sequence may encode variant amino acid sequences, but one of ordinary skill in the art can easily identify such variant sequences by reference to the amino acid sequences of the present invention. A "variant" nucleic acid sequence has substantial homology or substantial similarity to a reference nucleic acid sequence (or a fragment thereof). A nucleic acid sequence or fragment thereof is "substantially homologous" (or "substantially identical") to a reference sequence if, when optimally aligned (with appropriate nucleotide insertions or deletions) with the other nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 70%, 75%, 80%, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or more% of the nucleotide bases. Methods for homology determination of nucleic acid sequences are known in the art.

[0145] Alternatively, a "variant" nucleic acid sequence is substantially homologous with (or substantially identical to) a reference sequence (or a fragment thereof) if the "variant" and the reference sequence they are capable of hybridizing under stringent (e.g. highly stringent) hybridization conditions. Nucleic acid sequence hybridization will be affected by such conditions as salt concentration (e.g. NaCI), temperature, or organic solvents, in addition to the base composition, length of the complementary strands, and the number of nucleotide base mismatches between the hybridizing nucleic acids, as will be readily appreciated by those skilled in the art. Stringent temperature conditions are preferably employed, and generally include temperatures in excess of 30°C, typically in excess of 37°C and preferably in excess of 45°C. Stringent salt conditions will ordinarily be less than 1000 mM, typically less than 500 mM, and preferably less than 200 mM. The pH is typically between 7.0 and 8.3. The combination of parameters is much more important than any single parameter.

[0146] Methods of determining nucleic acid percentage sequence identity are known in the art. By way of example, when assessing nucleic acid sequence identity, a sequence having a defined number of contiguous nucleotides may be aligned with a nucleic acid sequence (having the same number of contiguous nucleotides) from the corresponding portion of a nucleic acid sequence of the present invention. Tools known in the art for determining nucleic acid percentage sequence identity include Nucleotide BLAST (as described below).

[0147] One of ordinary skill in the art appreciates that different species exhibit "preferential codon usage". As used herein, the term "preferential codon usage" refers to codons that are most frequently used in cells of a certain species, thus favouring one or a few representatives of the possible codons encoding each amino acid. For example, the amino acid threonine (Thr) may be encoded by ACA, ACC, ACG, or ACT, but in mammalian host cells ACC is the most commonly used codon; in other species, different codons may be preferential. Preferential codons for a particular host cell species can be introduced into the polynucleotides of the present invention by a variety of methods known in the art. Introduction of preferential codon sequences into recombinant DNA can, for example, enhance production of the protein by making protein translation more efficient within a particular cell type or species.

[0148] A "fragment" of a polynucleotide of interest comprises a series of consecutive nucleotides from the sequence of said full-length polynucleotide. By way of example, a "fragment" of a polynucleotide of interest may comprise (or consist of) at least 600 consecutive nucleotides from the sequence of said polynucleotide (e.g. at least 600, 650, 700, or 750 consecutive nucleic acid residues of said polynucleotide). Typically, a fragment as defined herein retains the same function as the full-length polynucleotide.

[0149] A "fragment" of a polypeptide of interest comprises a series of consecutive amino acid residues from the sequence of said full-length polypeptide. By way of example, a "fragment" of a polypeptide of interest may comprise (or consist of) at least 100 consecutive amino acid residues from the sequence of said polypeptide (e.g. at least 100, 150, 200, or 250 consecutive amino acid residues of said polypeptide). Typically, a fragment as defined herein retains the same function as the full-length polypeptide.

[0150] The terms "decrease", "reduced", "reduction", or "inhibit" are all used herein to mean a decrease by a statistically significant amount. The terms "reduce," "reduction" or "decrease" or "inhibit" typically means a decrease by at least 10% as compared to a reference level (e.g. the absence of a given treatment) and can include, for example, a decrease by at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99%, or more. As used herein, "reduction" or "inhibition" encompasses a complete inhibition or reduction as compared to a reference level. " Complete inhibition" is a 100% inhibition (i.e. abrogation) as compared to a reference level.

[0151] The terms "increased", "increase", "enhance", or "activate" are all used herein to mean an increase by a statically significant amount. The terms "increased", "increase", "enhance", or "activate" can mean an increase of at least 25%, at least 50% as compared to a reference level, for example an increase of at least about 50%, or at least about 75%, or at least about 80%, or at least about 90%, at least about 95%, or at least about 98%, or at least about 99%, or at least about 100%, or at least about 250% or more compared with a reference level, or at least about a 1.5-fold, or at least about a 2-fold, or at least about a 2.5-fold, or at least about a 3-fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10-fold increase, or any increase between 1.5-fold and 10-fold or greater as compared to a reference level. In the context of a yield or titre, an "increase" is an observable or statistically significant increase in such level.

[0152] The terms "individual", "subject", and "patient", are used interchangeably herein to refer to a mammalian subject for whom diagnosis, prognosis, disease monitoring, treatment, therapy, and / or therapy optimisation is desired. The mammal can be (without limitation) a human, non-human primate, mouse, rat, dog, cat, horse, or cow. In a preferred embodiment, the individual, subject, or patient is a human. An "individual" may be an adult, juvenile or infant. An "individual" may be male or female.

[0153] A "subject in need" of treatment for a particular condition can be an individual having that condition, diagnosed as having that condition, or at risk of developing that condition. A subject can be one who has been previously diagnosed with or identified as suffering from or having a condition in need of treatment or one or more complications or symptoms related to such a condition, and optionally, have already undergone treatment for a condition as defined herein or the one or more complications or symptoms related to said condition. Alternatively, a subject can also be one who has not been previously diagnosed as having a condition as defined herein or one or more or symptoms or complications related to said condition. For example, a subject can be one who exhibits one or more risk factors for a condition, or one or more or symptoms or complications related to said condition or a subject who does not exhibit risk factors.

[0154] As used herein, the term "healthy individual" refers to an individual or group of individuals who are in a healthy state, e.g. individuals who have not shown any symptoms of the disease, have not been diagnosed with the disease and / or are not likely to develop the disease e.g. malaria. Preferably said healthy individual(s) is not on medication affecting malaria and has not been diagnosed with any other disease. The one or more healthy individuals may have a similar sex, age, and / or body mass index (BMI) as compared with the test individual. Application of standard statistical methods used in medicine permits determination of normal levels of expression in healthy individuals, and significant deviations from such normal levels.

[0155] Herein the terms "control" and "reference population" are used interchangeably.

[0156] The term "pharmaceutically acceptable" as used herein means approved by a regulatory agency of the Federal or a state government, or listed in the U. S. Pharmacopeia, European Pharmacopeia or other generally recognized pharmacopeia.

[0157] The term "simultaneous" as used herein means that the agents are administered concurrently, i.e. at the same time.

[0158] The term "sequential" as used herein means that the agents are administered one after the other. The term "separately" as used herein means that the agents are administered at different times. Dynamic Light Scattering (DLS) is an analytical technique used to measure the size distribution of small particles, such as the VLPs of the invention. It works by analysing the fluctuations in the intensity of light scattered by these particles as they undergo Brownian motion. The rate of these fluctuations is related to the size of the particles, with smaller particles causing faster fluctuations. DLS is commonly used in fields like chemistry, biology, and materials science to characterize particle size and study the behaviour of colloidal systems. The skilled person would therefore easily be able to assess the size of the VLPs of the invention using DLS.

[0159] Electron microscopy (EM) is an imaging technique that uses a beam of electrons, rather than light, to magnify and visualize objects at very high resolutions, such as the VLPs of the invention. Due to the shorter wavelength of electrons compared to light, electron microscopy can reveal much finer structural details, making it possible to view objects at the nanoscale, such as the VLPs of the invention. There are two main types of electron microscopy: Transmission Electron Microscopy (TEM), which provides detailed images of thin slices of a specimen, and Scanning Electron Microscopy (SEM), which provides detailed surface images. Electron microscopy is widely used in fields like materials science, biology, and nanotechnology for detailed structural analysis. The skilled person would therefore easily be able to assess the size of the VLPs of the invention using EM.

[0160] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that such publications constitute prior art to the claims appended hereto.

[0161] Plasmodium vivax circumsporozoite protein (PvCSP)

[0162] Circumsporozoite protein (CSP) is a major surface protein of sporozoites. PfCSP contains a flexible N-terminal domain with a heparan sulfate binding site, a central region consisting of multiple NANP repeats, a structured C-terminal domain with a thrombospondin-like type I repeat (aTSR). However, unlike the P. falciparum, P. vivax CSP (PvCSP) is genetically diverse with two distinct genotypes or strains, VK210 (also known as Type I) and VK247 (also known as Type 2). The full-length VK210 PvCSP has an amino acid sequence according to GenBank Accession No. P08677 (version 2, updated 01 February 1996, accessed 05 September 2024 - SEQ ID NO: 70). The full-length VK247 PvCSP has an amino acid sequence according to GenBank Accession No. Q26166 (version 1, updated 1 November 1996, accessed 05 September 2024-SEQ ID NO: 71). Both of these sequences are herein incorporated by reference in their entirety.

[0163] The VK210 PvCSP and VK247 PvCSP differ in their central region, which consists of repeat units (also referred to interchangeably as repeat motifs) as shown in Figure 1 of Kucharska et al (eLife. 2022; 11: e72908), which is herein incorporated by reference in its entirety. This central region is known as the central repeat region. The central repeat region of VK210 PvCSP is defined by residues 96 to 275 of the GenBank Accession No. P08677 sequence (SEQ ID NO: 72). The central repeat region of VK247 PvCSP is defined by residues 6 to 193 of the GenBank Accession No. Q26166 sequence (SEQ ID NO: 73).

[0164] A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of repeat motifs of the PvCSP from a Type I (VK210) P. vivax. A PvCSP protein or immunogenic fragment thereof of the invention may comprise or consist of at least 10 (e.g. 10, 11, 12, 13, 14, 15, 20, 25 or more) repeat motifs from the central repeat region of a Type I (VK210) PvCSP.

[0165] The repeat motifs from the central repeat region of a Type I (VK210) PvCSP may be selected independently. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise any number of a Type I (VK210) PvCSP repeat motif of interest. In other words, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise a single copy of a Type I (VK210) PvCSP repeat motif, or may comprise multiple copies of said Type I (VK210) PvCSP repeat motif. By way of non-limiting, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise at least 5 copies of a first repeat motif from a Type I (VK210) PvCSP, at least five copies of a second repeat motif from a Type I (VK210) PvCSP and at least one copy of a third repeat motif from a Type I (VK210) PvCSP.

[0166] A repeat motif from a Type I (VK210) PvCSP may be selected from the group consisting of: GDRADGQPA (SEQ ID NO: 4); GDRAAGQPA (SEQ ID NO: 5); and GNGAGGQAA (SEQ ID NO: 6), or a variant thereof. A variant of SEQ ID NO: 4 may differ from SEQ ID NO: 4 by one or two amino acids, e.g. by substitution or deletion, typically substitution, preferably a conservative amino acid substitution. A variant of SEQ ID NO: 5 may differ from SEQ ID NO: 5 by one or two amino acids, e.g. by substitution or deletion, typically substitution, preferably a conservative amino acid substitution. A variant of SEQ ID NO: 6 may differ from SEQ ID NO: 6 by one or two amino acids, e.g. by substitution or deletion, typically substitution, preferably a conservative amino acid substitution.

[0167] A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of at least one copy (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of SEQ ID NO: 4 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of 5, 6, 7, 8, 9 or 10 copies of SEQ ID NO: 4 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise 6 copies of SEQ ID NO: 4 or a variant thereof.

[0168] Alternatively or in addition, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of at least one copy (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of SEQ ID NO: 5 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of 5, 6, 7, 8, 9 or 10 copies of SEQ ID NO: 5 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise 6 copies of SEQ ID NO: 5 or a variant thereof.

[0169] Further alternatively or in addition, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of at least one copy (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of SEQ ID NO: 6 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of 1, 2 or 3 copies of SEQ ID NO: 6 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise 1 copy of SEQ ID NO: 6 or a variant thereof.

[0170] A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (i) at least 5 copies of SEQ ID NO: 4 or a variant thereof; and (ii) at least 5 copies of SEQ ID NO: 5 or a variant thereof. A PvCSP protein or immunogenie fragment thereof according to the invention may comprise or consist of: (i) at least 5 copies of SEQ ID NO: 4; and (ii) at least 5 copies of SEQ ID NO: 5. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (i) 6 copies of SEQ ID NO: 4 or a variant thereof; and (ii) 6 copies of SEQ ID NO: 5 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (i) 6 copies of SEQ ID NO: 4; and (ii) 6 copies of SEQ ID NO: 5. A PvCSP protein or immunogenic fragment thereof according to the invention may further comprise or consist of at least one copy of SEQ ID NO: 6 or a variant thereof. Thus, a PvCSP protein or immunogenie fragment thereof according to the invention may comprise or consist of: (i) at least 5 copies of SEQ ID NO: 4 or a variant thereof; (ii) at least 5 copies of SEQ ID NO: 5 or a variant thereof; and (iii) at least one copy of SEQ ID NO: 6 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (i) at least 5 copies of SEQ ID NO: 4; (ii) at least 5 copies of SEQ ID NO: 5; and (iii) at least one copy of SEQ ID NO: 6. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (i) 6 copies of SEQ ID NO: 4 or a variant thereof; (ii) 6 copies of SEQ ID NO: 5 or a variant thereof; and (iii) one copy of SEQ ID NO: 6 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (i) 6 copies of SEQ ID NO: 4; (ii) 6 copies of SEQ ID NO: 5; and (iii) one copy of SEQ ID NO: 6.

[0171] The repeat motifs from the central repeat region of a Type I (VK210) PvCSP may be arranged in any order.

[0172] In embodiments wherein a PvCSP protein or immunogenic fragment thereof according to the invention comprises two or more repeat motifs from the central repeat region of a Type I (VK210) PvCSP, the copies of each repeat motif may be arranged contiguously. By way of non-limiting, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise 5 copies of a first repeat motif from a Type I (VK210) PvCSP [210RM1], five copies of a second repeat motif from a Type I (VK210) PvCSP ([210RM2]), and one copy of a third repeat motif from a Type I (VK210) PvCSP ([210RM3]). These may be arranged in any of the following non-limiting configurations: (i) [210RM1]5[210RM2]5[210RM3]; (ii) [210RM2]5[210RM1]5[210RM3]; (iii) [210RM1]5[210RM3][210RM2]5; (iv) [210RM2]5[210RM3][210RM1]5; (v) [210RM3][210RM1]5[210RM2]5; or (vi) [210RM3][210RM2]5[210RM1]5.

[0173] Wherein a PvCSP protein or immunogenic fragment thereof according to the invention comprises at least one copy at least two different Type I (VK210) PvCSP repeat motifs, the copies of the first repeat motif may be contiguous and the copies of the second motif may be contiguous. The contiguous copies of the first repeat motif may be N-terminal to the contiguous copies of the second motif. The contiguous copies of the second motif may be N-terminal to the contiguous copies of the first repeat motif.

[0174] Thus, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (i) at least 5 contiguous copies of SEQ ID NO: 4 or a variant thereof; and (ii) at least 5 contiguous copies of SEQ ID NO: 5 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (i) at least 5 contiguous copies of SEQ ID NO: 4; and (ii) at least 5 contiguous copies of SEQ ID NO: 5. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (i) 6 contiguous copies of SEQ ID NO: 4 or a variant thereof; and (ii) 6 contiguous copies of SEQ ID NO: 5 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (i) 6 contiguous copies of SEQ ID NO: 4; and (ii) 6 contiguous copies of SEQ ID NO: 5. The copies of SEQ ID NO: 4 may be N-terminal to the copies of SEQ ID NO: 5. The copies of SEQ ID NO: 5 may be N-terminal to the copies of SEQ ID NO: 4. Wherein a PvCSP protein or immunogenic fragment thereof according to the invention further comprises at least one copy of SEQ ID NO: 6, the contiguous nature of the copies of SEQ ID NO: 4 and / or SEQ ID NO: 5 may be retained.

[0175] Alternatively, in embodiments wherein a PvCSP protein or immunogenic fragment thereof according to the invention comprises two or more repeat motifs from the central repeat region of a Type I (VK210) PvCSP, the copies of each repeat motif may be interspersed. By interspersed, it is typically meant that each individual copy of a repeat motif remains intact / undisrupted, but that a copy of a different repeat motif is inserted between two copies of the repeat motif(s). A PvCSP protein or immunogenic fragment thereof according to the invention may comprise 5 copies of a first repeat motif from a Type I (VK210) PvCSP [210RM1], five copies of a second repeat motif from a Type I (VK210) PvCSP ([210RM2]) and one copy of a third repeat motif from a Type I (VK210) PvCSP ([210RM3]). These may be arranged in any of the following non-limiting configurations: (i) [210RMl]5[210RM2]2[210RM3][210RM2]3; (ii) [210RMl][210RM2]2[210RM3][210RMl]4[210RM2]3; (iii) [210RM1][210RM2][210RM1][210RM2][210RM3][210RM1]3[210RM2]3[210RM1]; or (iv) [210RMl]3[210RM2][210RMl]2[210RM2]2[210RMl][210RM2]3[210RM3]. Preferably, the repeat motifs from a Type I (VK210) PvCSP are arranged [210RMl]3[210RM2][210RMl]2[210RM2]2[210RMl][210RM2]3[210RM3].

[0176] Wherein a PvCSP protein or immunogenic fragment thereof according to the invention comprises at least one copy at least two different Type I (VK210) PvCSP repeat motifs, at least one copy of the second repeat motif may be interspersed with the copies of the first repeat motif. Alternatively or in addition, at least one copy of the first repeat motif may be interspersed with the copies of the second motif. Preferably, wherein a PvCSP protein or immunogenic fragment thereof according to the invention comprises at least one copy at least two different Type I (VK210) PvCSP repeat motifs, at least one copy of the second repeat motif is interspersed with the copies of the first repeat motif and at least one copy of the first repeat motif is interspersed with the copies of the second motif.

[0177] Thus, a PvCSP protein or immunogenie fragment thereof according to the invention may comprise or consist of: (a) at least 5 copies of SEQ ID NO: 4 or a variant thereof; and (b) at least 5 copies of SEQ ID NO: 5 ora variant thereof; wherein (i) at least one copy of SEQ ID NO: 5 or a variant thereof is interspersed between copies of SEQ ID NO: 4 or a variant thereof; and / or (ii) at least one copy of SEQ ID NO: 4 or a variant thereof is interspersed between copies of SEQ ID NO: 5 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) at least 5 copies of SEQ ID NO: 4 or a variant thereof; and (b) at least 5 copies of SEQ ID NO: 5 or a variant thereof; wherein (i) at least one copy of SEQ ID NO: 5 or a variant thereof is interspersed between copies of SEQ ID NO: 4 or a variant thereof; and (ii) at least one copy of SEQ ID NO: 4 or a variant thereof is interspersed between copies of SEQ ID NO: 5 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) at least 5 copies of SEQ ID NO: 4; and (b) at least 5 copies of SEQ ID NO: 5; wherein (i) at least one copy of SEQ ID NO: 5 is interspersed between copies of SEQ ID NO: 4; and / or (ii) at least one copy of SEQ ID NO: 4 is interspersed between copies of SEQ ID NO: 5. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) at least 5 copies of SEQ ID NO: 4; and (b) at least 5 copies of SEQ ID NO: 5; wherein (i) at least one copy of SEQ ID NO: 5 is interspersed between copies of SEQ ID NO: 4; and (ii) at least one copy of SEQ ID NO: 4 is interspersed between copies of SEQ ID NO: 5. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) 6 copies of SEQ ID NO: 4 or a variant thereof; and (b) 6 copies of SEQ ID NO: 5 or a variant thereof; wherein (i) at least one copy of SEQ ID NO: 5 or a variant thereof is interspersed between copies of SEQ ID NO: 4 or a variant thereof; and / or (ii) at least one copy of SEQ ID NO: 4 or a variant thereof is interspersed between copies of SEQ ID NO: 5 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) 6 copies of SEQ ID NO: 4; and (b) 6 copies of SEQ ID NO: 5 or a variant thereof; wherein (i) at least one copy of SEQ ID NO: 5 is interspersed between copies of SEQ ID NO: 4; and (ii) at least one copy of SEQ ID NO: 4 is interspersed between copies of SEQ ID NO: 5. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) 6 copies of SEQ ID NO: 4; and (b) 6 copies of SEQ ID NO: 5 or a variant thereof; wherein (i) one copy of SEQ ID NO: 5 is interspersed between copies of SEQ ID NO: 4; and (ii) one copy of SEQ ID NO: 4 is interspersed between copies of SEQ ID NO: 5. The interspersed copy of SEQ ID NO: 4 may be present between: (i) the 3rdand 4th; (ii) 1stand 2nd; (iii) 2ndand 3rd; (iv) 4thand 5th; or (v) 5thand 6th; contiguous copies of SEQ ID NO: 5. Preferably the interspersed copy of SEQ ID NO: 4 is present between the 3rdand 4thcontiguous copies of SEQ ID NO: 5. The interspersed copy of SEQ ID NO: 5 may be present between: (i) 2ndand 3rd; (ii) 1stand 2nd; (iii) the 3rdand 4th; (iv) 4thand 5th; or (v) 5thand 6th; contiguous copies of SEQ ID NO: 4. Preferably the interspersed copy of SEQ ID NO: 5 is present between the 2ndand 3rdcontiguous copies of SEQ ID NO: 4. The noninterspersed copies of SEQ ID NO: 4 may be N-terminal to the non-interspersed copies of SEQ ID NO: 5. The non-interspersed copies of SEQ ID NO: Smay be N-terminal to the non-interspersed copies of SEQ ID NO: 4. Wherein a PvCSP protein or immunogenic fragment thereof according to the invention further comprises at least one copy of SEQ ID NO: 6, the interspersed nature of the copies of SEQ ID NO: 4 and / or SEQ ID NO: 5may be retained. The at least one copy of SEQ ID NO: 6 may be C-terminal to the noninterspersed copies of SEQ ID NO: 4 and / or SEQ ID NO: 5. The at least one copy of SEQ ID NO: 6 may be N-terminal to the non-interspersed copies of SEQ ID NO: 4 and / or SEQ ID NO: 5.

[0178] A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to SEQ ID NO: I. The PvCSP protein or immunogenic fragment thereof may comprise an amino acid sequence that is at least 85% identical to SEQ ID NO: 1. The PvCSP protein or immunogenic fragment thereof may consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 1. The PvCSP protein or immunogenic fragment thereof may comprise an amino acid sequence that is at least 90% identical to SEQ ID NO: 1. The PvCSP protein or immunogenic fragment thereof may consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 1. The PvCSP protein or immunogenic fragment thereof may comprise an amino acid sequence that is at least 95% identical to SEQ ID NO: 1. The PvCSP protein or immunogenic fragment thereof may consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 1. The PvCSP protein or immunogenic fragment thereof may comprise an amino acid sequence that is at least 99% identical to SEQ ID NO: 1. The PvCSP protein or immunogenic fragment thereof may consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 1. The PvCSP protein or immunogenic fragment thereof may comprise the amino acid sequence of SEQ ID NO: 1. The PvCSP protein or immunogenic fragment thereof may consist of the amino acid sequence of SEQ ID NO: 1.

[0179] Alternatively or in addition, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of repeat motifs of the PvCSP from a Type II (VK247) P. vivax. A PvCSP protein or immunogenic fragment thereof of the invention may comprise or consist of at least 10 (e.g. 10, 11, 12, 13, 14, 15, 20, 25 or more) repeat motifs from the central repeat region of a Type II (VK247) PvCSP.

[0180] The repeat motifs from the central repeat region of a Type II (VK247) PvCSP may be selected independently. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise any number of a Type II (VK247) PvCSP repeat motif of interest. In other words, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise a single copy of a Type II (VK247) PvCSP repeat motif, or may comprise multiple copies of said Type II (VK247) PvCSP repeat motif. By way of non-limiting, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise at least 5 copies of a first repeat motif from a Type II (VK247) PvCSP, at least one copy of a second repeat motif from a Type II (VK247) PvCSP and at least one copy of a third repeat motif from a Type II (VK247) PvCSP.

[0181] A repeat motif from a Type II (VK247) PvCSP may be selected from the group consisting of: ANGAGNQPG (SEQ ID NO: 7); EDGAGNQPG (SEQ ID NO: 8); NGAGDQPG (SEQ ID NO: 9); ANGADDQPG (SEQ ID NO: 10); ANGAGGQAA (SEQ ID NO: 11); and GGNA ANKKAEDA GGNA GGNA (SEQ ID NO: 12). A variant of SEQ ID NO: 7 may differ from SEQ ID NO: 7 by one or two amino acids, e.g. by substitution or deletion, typically substitution, preferably a conservative amino acid substitution. A variant of SEQ ID NO: 8 may differ from SEQ ID NO: 8 by one or two amino acids, e.g. by substitution or deletion, typically substitution, preferably a conservative amino acid substitution. A variant of SEQ ID NO: 9 may differ from SEQ ID NO: 9 by one or two amino acids, e.g. by substitution or deletion, typically substitution, preferably a conservative amino acid substitution. A variant of SEQ ID NO: 10 may differ from SEQ ID NO: 10 by one or two amino acids, e.g. by substitution or deletion, typically substitution, preferably a conservative amino acid substitution. A variant of SEQ ID NO: 11 may differ from SEQ ID NO: 11 by one or two amino acids, e.g. by substitution or deletion, typically substitution, preferably a conservative amino acid substitution. A variant of SEQ ID NO: 12 may differ from SEQ ID NO: 12 by one, two, three or four amino acids, preferably one or two amino acids, e.g. by substitution or deletion, typically substitution, preferably a conservative amino acid substitution.

[0182] A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of at least one copy (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of SEQ ID NO: 7 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of 5, 6, 7, 8, 9 or 10 copies of SEQ ID NO: 7 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise 8 copies of SEQ ID NO: 7 or a variant thereof.

[0183] Alternatively or in addition, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of at least one copy (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of SEQ ID NO: 8 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of 1, 2 or 3 copies of SEQ ID NO: 8 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise 1 copy of SEQ ID NO: 8 or a variant thereof.

[0184] Further alternatively or in addition, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of at least one copy (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of SEQ ID NO: 9 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of 1, 2 or 3 copies of SEQ ID NO: 9 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise 2 copies of SEQ ID NO: 9 or a variant thereof.

[0185] Further alternatively or in addition, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of at least one copy (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of SEQ ID NO: 10 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of 1, 2 or 3 copies of SEQ ID NO: 10 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise 1 copy of SEQ ID NO: 10 or a variant thereof.

[0186] Further alternatively or in addition, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of at least one copy (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of SEQ ID NO: 11 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of 1, 2 or 3 copies of SEQ ID NO: 11 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise 1 copy of SEQ ID NO: 11 or a variant thereof.

[0187] Further alternatively or in addition, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of at least one copy (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) of SEQ ID NO: 12 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of 1, 2 or 3 copies of SEQ ID NO: 12 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise 1 copy of SEQ ID NO: 12 or a variant thereof.

[0188] A PvCSP protein or immunogenic fragment thereof according to the invention may comprise at least 1 copy of any two, three, four, five or all six of SEQ ID NO: 8, 9, 10, 11, or 12, or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise at least 1 copy of any two of SEQ ID NO: 8, 9, 10, 11, or 12, or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise at least 1 copy of any three of SEQ ID NO: 8, 9, 10, 11, or 12, or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise at least 1 copy of any four of SEQ ID NO: 8, 9, 10, 11, or 12, or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise at least 1 copy of all five of SEQ ID NO: 8, 9, 10, 11, or 12, or a variant thereof.

[0189] Thus, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) at least 5 copies of SEQ ID NO: 7 or a variant thereof; and (b) at least 1 copy of any two or more of (i) SEQ ID NO: 8 or a variant thereof; (ii) SEQ ID NO: 9 or a variant thereof; (iii) SEQ ID NO: 10 or a variant thereof; (iv) SEQ ID NO: 11 or a variant thereof; or (v) SEQ ID NO: 12 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) at least 5 copies of SEQ ID NO: 7 or a variant thereof; and (b) at least 1 copy of (i)+(ii), (i)+(iii), (i)+(iv), (i)+(v), (ii)+(iii), (ii)+(iv), (ii)+(v), (iii)+(iv), (iii)+(v), (iv)+(v), (i)+(ii)+(iii), (i)+(ii)+(iv), (i)+(ii)+(v), (i)+(iii)+(iv), (i)+(iii)+(v), (i)+(iv)+(v), (ii)+(iii)+(iv), (ii)+(iii)+(v), (ii)+(iv)+(v), (iii)+(iv)+(v), (i)+(ii)+(iii)+(iv), (i)+(ii)+(iii)+(v), (i)+(ii)+(iv)+(v), (i)+(iii)+(iv)+(v), (ii)+(iii)+(iv)+(v) or(i)+(ii)+(iii)+(iv)+(v).

[0190] A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) at least 5 copies of SEQ ID NO: 7; and (b) at least 1 copy of any two or more of (i) SEQ ID NO: 8; (ii) SEQ ID NO: 9; (iii) SEQ ID NO: 10; (iv) SEQ ID NO: 11; or (v) SEQ ID NO: 12. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) at least 5 copies of SEQ ID NO: 7 or a variant thereof; and (b) at least 1 copy of (i)+(ii), (i)+(iii), (i)+(iv), (i)+(v), (ii)+(iii), (ii)+(iv), (ii)+(v), (iii)+(iv), (iii)+(v), (iv)+(v), (i)+(ii)+(iii), (i)+(ii)+(iv), (i)+(ii)+(v), (i)+(iii)+(iv), (i)+(iii)+(v), (i)+(iv)+(v), (ii)+(iii)+(iv), (ii)+(iii)+(v), (ii)+(iv)+(v), (iii)+(iv)+(v), (i)+(ii)+(iii)+(iv), (i)+(ii)+(iii)+(v), (i)+(ii)+(iv)+(v), (i)+(iii)+(iv)+(v), (ii)+(iii)+(iv)+(v) or(i)+(ii)+(iii)+(iv)+(v).

[0191] A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) 8 copies of SEQ ID NO: 7 or a variant thereof; and (b) 1 copy of any two or more of (i) SEQ ID NO: 8 or a variant thereof; (ii) SEQ ID NO: 9 or a variant thereof; (iii) SEQ ID NO: 10 or a variant thereof; (iv) SEQ ID NO: 11 or a variant thereof; or (v) SEQ ID NO: 12 or a variant thereof. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) 8 copies of SEQ ID NO: 7 or a variant thereof; and (b) 1 copy of (i)+(ii), (i)+(iii), (i)+(iv), (i)+(v), (ii)+(iii), (ii)+(iv), (ii)+(v), (iii)+(ivh (iii)+(v), (iv)+(v), (i)+(ii)+(iii), (i)+(ii)+(ivL (i)+(ii)+(vL (i)+(iii)+(iv), (i)+(iii)+(v), (i)+(iv)+(v), (ii)+(iii)+(iv), (ii)+(iii)+(v), (ii)+(iv)+(v), (iii)+(iv)+(v), (i)+(ii)+(iii)+(iv), (i)+(ii)+(iii)+(v), (i)+(ii)+(iv)+(v), (i)+(iii)+(iv)+(v), (ii)+(iii)+(iv)+(v) or(i)+(ii)+(iii)+(iv)+(v).

[0192] A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) 8 copies of SEQ ID NO: 7; and (b) 1 copy of any two or more of (i) SEQ ID NO: 8; (ii) SEO ID NO: 9; (iii) SEQ ID NO: 10; (iv) SEQ ID NO: 11; or (v) SEQ ID NO: 12. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) 8 copies of SEQ ID NO: 7 or a variant thereof; and (b) 1 copy of (i)+(ii), (i)+(iii), (i)+(iv), (i)+(v), (ii)+(iii), (ii)+(iv), (ii)+(v), (iii)+(iv), (iii)+(v), (iv)+(v), (i)+(ii)+(iii), (i)+(ii)+(iv), (i)+(ii)+(v);(i)+(iii)+(iv), (i)+(iii)+(v), (i)+(iv)+(v), (ii)+(iii)+(iv), (ii)+(Hi)+(v), (ii)+(iv)+(v), (iii)+(iv)+(v), (i)+(ii)+(iii)+(iv), (i)+(ii)+(iii)+(v), (i)+(ii)+(iv)+(v), (i)+(iii)+(iv)+(v), (ii)+(iii)+(iv)+(v) or(i)+(ii)+(iii)+(iv)+(v).

[0193] The repeat motifs from the central repeat region of a Type II (VK247) PvCSP may be arranged in any order.

[0194] In embodiments wherein a PvCSP protein or immunogenic fragment thereof according to the invention comprises two or more repeat motifs from the central repeat region of a Type II (VK247) PvCSP, the copies of each repeat motif may be arranged contiguously. By way of non-limiting, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise 8 copies of a first repeat motif from a Type II (VK247) PvCSP ([247RM1]), one copy of a second repeat motif from a Type II (VK247) PvCSP ([247RM2]) and one copy of a third repeat motif from a Type II (VK247) PvCSP ([247RM3]). These may be arranged in any of the following configurations: (i) [247RMl]s[247RIVI2][247RM3]; (ii) [247RM2][247RMl]g[247RM3]; (iii) [247RM2][247RM3][247RM1]8; (iv) [247RM1]8[247RM3][247RM2]; (v) [247RM3][247RMl]g[247RM2]; or (vi) [247RM3][247RM2][247RM1]8.

[0195] Wherein a PvCSP protein or immunogenic fragment thereof according to the invention comprises at least one copy at least two different Type II (VK247) PvCSP repeat motifs, at least two copies of the first repeat motif may be contiguous. The contiguous copies of the first repeat motif may be N-terminal to the copies of the second motif. The second motif may be N-terminal to the contiguous copies first repeat motif.

[0196] Thus, a PvCSP protein or immunogenie fragment thereof according to the invention may comprise or consist of: (a) at least 5 copies of SEQ ID NO: 7 or a variant thereof, at least three of which are contiguous; and (b) at least 1 copy of any two or more of (i) SEQ ID NO: 8 or a variant thereof; (ii) SEQ ID NO: 9 or a variant thereof; (iii) SEQ ID NO: 10 or a variant thereof; (iv) SEQ ID NO: 11 or a variant thereof; or (v) SEQ ID NO: 12 or a variant thereof, as described elsewhere herein. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) at least 5 copies of SEQ ID NO: 7, at least three of which are contiguous; and (b) at least 1 copy of any two or more of (i) SEQ ID NO: 8; (ii) SEQ ID NO: 9; (iii) SEQ ID NO: 10; (iv) SEQ ID NO: 11; or (v) SEQ ID NO: 12, as described elsewhere herein. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) 8 copies of SEQ ID NO: 7 or a variant thereof, at least three of which are contiguous; and (b) I copy of any two or more of (i) SEQ ID NO: 8 or a variant thereof; (ii) SEQ ID NO: 9 or a variant thereof; (iii) SEQ ID NO: 10 or a variant thereof; (iv) SEQ ID NO: 11 or a variant thereof; or (v) SEQ ID NO: 12 or a variant thereof, as described elsewhere herein. A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) 8 copies of SEQ ID NO: 7, at least three of which are contiguous; and (b) 1 copy of any two or more of (i) SEQ ID NO: 8; (ii) SEQ ID NO: 9; (iii) SEQ ID NO: 10; (iv) SEQ ID NO: 11; or (v) SEQ ID NO: 12, as described elsewhere herein.

[0197] The contiguous copies of SEQ ID NO: 7 or a variant thereof may be N-terminal to any two or more of (i) SEQ ID NO: 8; (ii) SEQ ID NO: 9; (iii) SEQ ID NO: 10; (iv) SEQ ID NO: 11; or (v) SEQ ID NO: 12, or variants thereof as described elsewhere herein. The contiguous copies of SEQ ID NO: 7 or a variant thereof may be C-terminal to any two or more of (i) SEQ ID NO: 8; (ii) SEQ ID NO: 9; (iii) SEQ ID NO: 10; (iv) SEQ ID NO: 11; or (v) SEQ ID NO: 12, or variants thereof as described elsewhere herein.

[0198] Alternatively, in embodiments wherein a PvCSP protein or immunogenic fragment thereof according to the invention comprises two or more repeat motifs from the central repeat region of a Type II (VK247) PvCSP, the copies of each repeat motif may be interspersed. By interspersed, it is typically meant that each individual copy of a repeat motif remains intact / undisrupted, but that a copy of a different repeat motif is inserted between two copies of the repeat motif(s). By way of non-limiting, a PvCSP protein or immunogenic fragment thereof according to the invention may comprise 8 copies of a first repeat motif from a Type II (VK247) PvCSP [247RM1], one copy of a second repeat motif from a Type II (VK247) PvCSP ([247RM2]), one copy of a third repeat motif from a Type II (VK247) PvCSP ([247RM3]), one copy of a fourth repeat motif from a Type II (VK247) PvCSP ([247RM4]), one copy of a fifth repeat motif from a Type II (VK247) PvCSP ([247RM5]), and two copies of a sixth repeat motif from a Type II (VK247) PvCSP ([247RM6]). These may be arranged in any of the following non-limiting configurations: (i) [247RM2][247RM1]3[247RM6][247RM1][247RM6][247RM1]2[247RM3][247RM1]2[247RM4][247RM5]; (ii) [247RM1]3[247RM3][247RM6][247RM1][247RM6][247RM1]2[247RM2][247RM1]2[247RM4] [247RM5]; or (iii) [247RM2][247RM1]3[247RM6][247RM1][247RM6][247RM1]2[247RM3][247RM1]2[247RM4][247RM5]. Preferably, the repeat motifs from a Type II (VK247) PvCSP are arranged [247RM2][247RM1]3[247RM6][247RM1][247RM6][247RM1]2[247RM3][247RM1]2[247RM4][247RM5].

[0199] Wherein a PvCSP protein or immunogenic fragment thereof according to the invention comprises at least one copy at least two different Type II (VK247) PvCSP repeat motifs, at least one copy of the second repeat motif may be interspersed with the copies of the first repeat motif. Alternatively or in addition, at least one copy of the first repeat motif may be interspersed with the copies of the second motif. Wherein a PvCSP protein or immunogenic fragment thereof according to the invention comprises at least one copy at least two different Type II (VK247) PvCSP repeat motifs, at least one copy of the second repeat motif is interspersed with the copies of the first repeat motif and at least one copy of the first repeat motif is interspersed with the copies of the second motif.

[0200] A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to SEQ ID NO: 2. The PvCSP protein or immunogenic fragment thereof may comprise an amino acid sequence that is at least 85% identical to SEQ ID NO: 2. The PvCSP protein or immunogenic fragment thereof may consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 2. The PvCSP protein or immunogenic fragment thereof may comprise an amino acid sequence that is at least 90% identical to SEQ ID NO: 2. The PvCSP protein or immunogenic fragment thereof may consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 2. The PvCSP protein or immunogenic fragment thereof may comprise an amino acid sequence that is at least 95% identical to SEQ ID NO: 2. The PvCSP protein or immunogenic fragment thereof may consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 2. The PvCSP protein or immunogenic fragment thereof may comprise an amino acid sequence that is at least 99% identical to SEQ ID NO: 2. The PvCSP protein or immunogenic fragment thereof may consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 2. The PvCSP protein or immunogenic fragment thereof may comprise the amino acid sequence of SEQ ID NO: 2. The PvCSP protein or immunogenic fragment thereof may consist of the amino acid sequence of SEQ ID NO: 2.

[0201] A PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of repeat motifs of the PvCSP from both a Type I (VK210) P. vivax and a Type II (VK247) P. vivax. Such PvCSP proteins and immunogenic fragments may be referred to as chimeric PvCSP proteins and chimeric PvCSP immunogenic fragments.

[0202] Thus, the invention provides a polypeptide comprising or consisting of: (a) at least 10 repeat motifs derived from the central repeat region of a Type I (VK210) of P. vivax; and / or (b) at least 10 repeat motifs derived from the central repeat region of a Type II (VK247) of P. vivax. Said polypeptide may be referred to as a chimeric PvCSP protein or immunogenic fragment thereof.

[0203] The repeat motifs of the PvCSP from a Type I (VK210) P. vivax and the repeat motifs of the PvCSP from a Type II (VK247) P. vivax may be selected independently from each other. Any repeat motif(s) from a Type I (VK210) PvCSP protein or immunogenic fragment thereof as described herein may be used in combination with any repeat motif(s) from a Type II (VK247) PvCSP protein or immunogenic fragment thereof. Thus, all the disclosure herein in relation to repeat motifs of the PvCSP from a Type I (VK210) P. vivax. applies equally and without reservation in the context of PvCSP proteins or immunogenic fragments thereof according to the invention which comprise repeat motifs from both Type I (VK210) PvCSP and Type II (VK247) PvCSP (chimeric PvCSP proteins or immunogenic fragments thereof). Similarly, all the disclosure herein in relation to repeat motifs of the PvCSP from a Type II (VK247) P. vivax. applies equally and without reservation in the context of PvCSP proteins or immunogenic fragments thereof according to the invention which comprise repeat motifs from both Type I (VK210) PvCSP and Type II (VK247) PvCSP (chimeric PvCSP proteins or immunogenic fragments thereof).

[0204] The configuration of the set of repeat motifs from the Type I (VK210) PvCSP repeat motifs and the set of repeat motifs from the Type II (VK247) PvCSP may be as described herein. Thus, the repeat motifs from the Type I (VK210) PvCSP may be contiguous or interspersed, as described herein. Similarly, the repeat motifs from the Type II (VK247) PvCSP may be contiguous or interspersed, as described herein.

[0205] Typically, the chimeric PvCSP protein or immunogenic fragment thereof comprises a first domain comprising or consisting of the repeat motifs from the Type I (VK210) PvCSP and a second domain comprising or consisting of repeat motifs from the Type II (VK247) PvCSP. The first domain may be N-terminal to the second domain. In other words, the chimeric PvCSP protein or immunogenic fragment thereof may have the structure N'-[Type I (VK210) PvCSP repeat motifs]-[Type II (VK247) PvCSP repeat motifs]-C', wherein within each domain the Type I or Type II motifs can be independently contiguous or interspersed as described herein. Alternatively, the first domain may beC-terminal to the second domain. In other words, the chimeric PvCSP protein or immunogenic fragment thereof may have the structure N'-[Type II (VK247) PvCSP repeat motifs]-[Type I (VK210) PvCSP repeat motifs]-C', wherein within each domain the Type I or Type II motifs can be independently contiguous or interspersed as described herein. Preferably, the first domain is N-terminal to the second domain, such that the polypeptides have the structure N'-[Type I (VK210) PvCSP repeat motifs]-[Type II (VK247) PvCSP repeat motifs]-C', wherein within each domain the Type I or Type II motifs can be independently contiguous or interspersed as described herein.

[0206] The first domain and the second domain may be covalently linked (e.g., fused) by recombinant, chemical or other suitable methods. The first and second domains may be linked directly or indirectly. Wherein the first domain and second domain are linked indirectly, this may be via a linker, such as a flexible linker. Examples of linkers, particularly peptide linkers are described herein. The structure of a polypeptide comprising a linker may be N'-[Type I (VK210) PvCSP repeat motifs]-linker-[Type II (VK247) PvCSP repeat motifs]-C' or N'-[Type II (VK247) PvCSP repeat motifs]-linker-[Type I (VK210) PvCSP repeat motifs]-C'. In some preferred embodiments, the first and second domain are linked directly, such that the C-terminal residue of the N-terminal domain is linked directly (e.g. via a peptide bond) to the N-terminal residue of the C-terminal domain.

[0207] By way of non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4 and at least 5 copies of the amino acid sequence of SEQ. ID NO: 5; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of any two or more of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0208] By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4 and at least 5 copies of the amino acid sequence of SEQ ID NO: 5; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of any three or more of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0209] By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4 and at least 5 copies of the amino acid sequence of SEQ ID NO: 5; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of any four or more of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0210] By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4 and at least 5 copies of the amino acid sequence of SEQ ID NO: 5; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of each of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0211] By way of non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4, at least 5 copies of the amino acid sequence of SEQ ID NO: 5, and at least one copy of SEQ ID NO: 6; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of any two or more of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0212] By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4, at least 5 copies of the amino acid sequence of SEQ ID NO: 5, and at least one copy of SEQ ID NO: 6; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of any three or more of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0213] By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4, at least 5 copies of the amino acid sequence of SEQ ID NO: 5, and at least one copy of SEQ ID NO: 6; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of any four or more of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0214] By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4, at least 5 copies of the amino acid sequence of SEQ ID NO: 5, and at least one copy of SEQ ID NO: 6; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of each of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5. By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4 and at least 5 copies of the amino acid sequence of SEQ ID NO: 5; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of any two or more of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (Hi) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0215] By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4 and at least 5 copies of the amino acid sequence of SEQ ID NO: 5; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of any three or more of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0216] By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4 and at least 5 copies of the amino acid sequence of SEQ ID NO: 5; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of any four or more of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0217] By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4 and at least 5 copies of the amino acid sequence of SEQ ID NO: 5; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of each of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0218] By way of non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4, at least 5 copies of the amino acid sequence of SEQ ID NO: 5, and at least one copy of SEQ ID NO: 6; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of any two or more of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0219] By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4, at least 5 copies of the amino acid sequence of SEQ ID NO: 5, and at least one copy of SEQ ID NO: 6; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of any three or more of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0220] By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4, at least 5 copies of the amino acid sequence of SEQ ID NO: 5, and at least one copy of SEQ ID NO: 6; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of any four or more of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0221] By way of a further non-limiting example, a chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of: (a) at least 5 copies of the amino acid sequence of SEQ ID NO: 4, at least 5 copies of the amino acid sequence of SEQ ID NO: 5, and at least one copy of SEQ ID NO: 6; and (b) at least 5 copies of the amino acid sequence of SEQ ID NO: 7 and at least one copy of each of (i) the amino acid sequence of SEQ ID NO: 8; (ii) the amino acid sequence of SEQ ID NO: 9; (iii) the amino acid sequence of SEQ ID NO: 10; (iv) the amino acid sequence of SEQ ID NO: 11; and (v) the amino acid sequence of SEQ ID NO: 12. At least one copy of SEQ ID NO: 5 may be interspersed between copies of SEQ ID NO: 4, optionally between the 3rdand 4thcontiguous copy of SEQ ID NO: 4. At least one copy of SEQ ID NO: 4 may be interspersed between copies of SEQ ID NO: 5, optionally between the 2ndand 3rdcontiguous copy of SEQ ID NO: 5.

[0222] A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to SEQ ID NO: 1; and (b) an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to SEQ ID NO: 2. Typically (a) is N-terminal to (b).

[0223] A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) an amino acid sequence that is at least 85% identical to SEQ ID NO: 1; and (b) an amino acid sequence that is at least 85% identical to SEQ ID NO: 2. Typically (a) is N-terminal to (b). A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) an amino acid sequence that is at least 90% identical to SEQ ID NO: 1; and (b) an amino acid sequence that is at least 90% identical to SEQ ID NO: 2. Typically (a) is N-terminal to (b). A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) an amino acid sequence that is at least 95% identical to SEQ ID NO: 1; and (b) an amino acid sequence that is at least 95% identical to SEQ ID NO: 2. Typically (a) is N-terminal to (b). A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) an amino acid sequence that is at least 99% identical to SEQ ID NO: 1; and (b) an amino acid sequence that is at least 99% identical to SEQ ID NO: 2. Typically (a) is N-terminal to (b). A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of: (a) the amino acid sequence of SEQ ID NO: 1; and (b) the amino acid sequence of SEQ ID NO: 2. Typically (a) is N-terminal to (b).

[0224] A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to SEQ ID NO: 3, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 85% identical to SEQ ID NO: 3, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 3, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 90% identical to SEQ ID NO: 3, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 3, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 95% identical to SEQ ID NO: 3, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 3, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof) according to the invention may comprise an amino acid sequence that is at least 99% identical to SEQ ID NO: 3, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 3, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise the amino acid sequence of SEQ ID NO: 3, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may consist of the amino acid sequence of SEQ ID NO: 3, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3.

[0225] A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 3. A chimeric PvCSP protein or immunogenic fragment thereof according to the invention may comprise or consist of the amino acid sequence of SEQ ID NO: 3.

[0226] A PvCSP protein or immunogenic fragment thereof according to the invention, particularly a chimeric PvCSP protein or immunogenic fragment thereof, may further comprise a PvCSP protein C-terminal region (CTR) or immunogenic fragment thereof. A PvCSP protein C-terminal region (CTR) of the invention may be interchangeably referred to as a CTR herein. A CTR or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to SEQ ID NO: 90, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 85% identical to SEQ ID NO: 90, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 90, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 90% identical to SEQ ID NO: 90, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 90, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 95% identical to SEQ ID NO: 90, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 90, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 99% identical to SEQ ID NO: 90, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 90, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may comprise the amino acid sequence of SEQ ID NO: 90, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may consist of the amino acid sequence of SEQ ID NO: 90, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 90.

[0227] A CTR or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 90. A CTR or immunogenic fragment thereof according to the invention may comprise or consist of the amino acid sequence of SEQ ID NO: 90. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of: (a) an amino acid sequence that is at least 85% identical to SEQ ID NO: 3; and (b) an amino acid sequence that is at least 85% identical to SEQ ID NO: 90. Typically (a) is N-terminal to (b). A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of: (a) an amino acid sequence that is at least 90% identical to SEQ ID NO: 3; and (b) an amino acid sequence that is at least 90% identical to SEQ ID NO: 90. Typically (a) is N-terminal to (b). A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of: (a) an amino acid sequence that is at least 95% identical to SEQ ID NO: 3; and (b) an amino acid sequence that is at least 95% identical to SEQ ID NO: 90. Typically (a) is N-terminal to (b). A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of: (a) an amino acid sequence that is at least 99% identical to SEQ ID NO: 3; and (b) an amino acid sequence that is at least 99% identical to SEQ ID NO: 90. Typically (a) is N-terminal to (b). A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of: (a) the amino acid sequence of SEQ ID NO: 3; and (b) the amino acid sequence of SEQ ID NO: 90. Typically (a) is N-terminal to (b).

[0228] A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to SEQ ID NO: 91, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 85% identical to SEQ ID NO: 91, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 91, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 90% identical to SEQ ID NO: 91, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 91, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 95% identical to SEQ ID NO: 91, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 91, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 99% identical to SEQ ID NO: 91, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 91, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise the amino acid sequence of SEQ ID NO: 91, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of the amino acid sequence of SEQ ID NO: 91, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 91.

[0229] A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 91. A chimeric PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of the amino acid sequence of SEQ ID NO: 91.

[0230] A PvCSP or immunogenic fragment thereof according to the invention, particularly a chimeric PvCSP protein or immunogenic fragment thereof, may comprise other components, such as a signal peptide, a linker, and / or label such as those described herein in the context of fusion proteins of the invention.

[0231] Accordingly, a PvCSP or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to any one of SEQ ID NO: 56 [SP2-PvCSP], SEQ ID NO: 57 [SP2-PvCSP-G4Sx3], SEQ ID NO: 58 [SP2-PvCSP-G4Sx3 - 4T], SEQ ID NO: 59 [SP2-PvCSP-EAAAKx3], SEQ ID NO: 60 [SP2-PvCSP- EAAAKx3 - 4T], SEQ ID NO: 61 [SP2-PvCSP- G4Sx3 - 4T - histag], SEQ ID NO: 62 [SP2-PvCSP- EAAAKx3 - 4T-histag] or 84 [PVCSP-GSGX2-4T-6XHis]. A PvCSP or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 56 to 62 or 84. A PvCSP or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 56 to 62 or 84. A PvCSP or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 56 to 62 or 84. A PvCSP or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 56 to 62 or 84. A PvCSP or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 56 to 62 or 84. A PvCSP or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 56 to 62 or 84. A PvCSP or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 99% identical to any one of SEQ ID NOs: 56 to 62 or 84. A PvCSP or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 99% identical to any one of SEQ ID NOs: 56 to 62 or 84. A PvCSP or immunogenic fragment thereof according to the invention may comprise the amino acid sequence of any one to any one of SEQ ID NOs: 56 to 62 or 84. A PvCSP or immunogenic fragment thereof according to the invention may consist of the amino acid sequence of any one to any one of SEQ ID NOs: 56 to 62 or 84.

[0232] Accordingly, a PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to any one of SEQ ID NO: 111 [SP2-PvCSP-CTR], 112 [SP2-PvCSP-CTR- GSx4], 113 [SP2-PvCSP- CTR-GSx4 - 4T] or 114 [SP2-PvCSP-CTR - GSx4 - 4T-Histag], A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 111 to 114. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 111 to 114. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 111 to 114. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 111 to 114. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 111 to 114. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 111 to 114. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 99% identical to any one of SEQ ID NOs: 111 to 114. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 99% identical to any one of SEQ ID NOs: 111 to 114. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise the amino acid sequence of any one to any one of SEQ ID NOs: 111 to 114. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of the amino acid sequence of any one to any one of SEQ ID NOs: 111 to 114.

[0233] A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to any one of SEQ ID NO: 56 to 62, 84, or 111. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 56 to 62, 84, or 111. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 56 to 62, 84, or 111. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 56 to 62, 84, or 111. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 56 to 62, 84, or 111. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 56 to 62, 84, or 111. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 56 to 62, 84, or 111. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 99% identical to any one of SEQ ID NOs: 56 to 62, 84, or 111. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 99% identical to any one of SEQ ID NOs: 56 to 62, 84, or 111. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may comprise the amino acid sequence of any one to any one of SEQ ID NOs: 56 to 62, 84, or 111. A PvCSP protein or immunogenic fragment thereof comprising a CTR or immunogenic fragment thereof according to the invention may consist of the amino acid sequence of any one to any one of SEQ ID NOs: 56 to 62, 84, or 111.

[0234] A PvCSP or immunogenic fragment thereof according to the invention may by glycosylated or unglycosylated, preferably unglycosylated. A PvCSP or immunogenic fragment thereof according to the invention may lack N-glycosylation. A PvCSP or immunogenic fragment thereof according to the invention may lack O-glycosylation. A PvCSP or immunogenic fragment thereof according to the invention may lack both N-glycosylation and O-glycosylation.

[0235] The invention also relates to fusion proteins comprising a PvCSP or immunogenic fragment thereof according to the invention, particularly a chimeric PvCSP protein or immunogenic fragment thereof, as well as to VLPs comprising a PvCSP or immunogenic fragment thereof, particularly a chimeric PvCSP protein or immunogenic fragment thereof, according to the invention. Such fusion proteins and VLPs are described further herein. For the avoidance of doubt, any and all disclosure herein in relation to PvCSP and immunogenic fragment thereof according to the invention, particularly chimeric PvCSP protein or immunogenic fragment thereof, applies equally and unreservedly to other aspects of the invention, such as fusion proteins and VLPs comprising a PvCSP or immunogenic fragment thereof according to the invention, particularly a chimeric PvCSP protein or immunogenic fragment thereof.

[0236] Plasmodium vivax 230 kDa sexual-stage protein (Pvs230)

[0237] Pfs230 is a 230 kDa sexual-stage protein that is composed of fourteen 6-cysteine-rich domains. Pfs230 is necessary for gamete fusion. Pfs230 is expressed in both male and female gametocytes without a known membrane anchor and appears on the surface of gametes as a complex with Pfs48 / 45, a glycosylphosphatidylinositol (GPI) anchored protein. Pvs230 is an ortholog of Pfs230.

[0238] The full-length Pvs230 has an amino acid sequence according to NCBI Accession No. XP_001613020.1 (version 1, updated 17 April 2018, accessed 17 August 2024). This sequence is herein incorporated by reference in its entirety.

[0239] The invention relates to the use of fragments of Pvs230. In particular, the invention relates to the use of fragments of Pv230 comprising or consisting of amino acid residues 226 to 427 of the Pvs230 amino acid sequence according to NCBI Accession No. XP 001613020.1 as described herein, or variants thereof. The amino acid sequence of residues 226 to 427 of the Pvs230 amino acid sequence according to NCBI Accession No. XP 001613020.1 is SEQ ID NO: 13 herein. An exemplary nucleic acid sequence encoding SEQ ID NO: 13 is provided by SEQ ID NO: 76.

[0240] The invention therefore relates to the use of SEQ ID NO: 13 and fragments and variants thereof. A variant of SEQ ID NO: 13 may differ from SEQ ID NO: 13 by no more than 10 amino acids, such as no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid; e.g. by substitution or deletion, typically substitution, preferably a conservative amino acid substitution.

[0241] A Pvs230 or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to SEQ ID NO: 13, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 85% identical to SEQ ID NO: 13, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 13, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 90% identical to SEQ ID NO: 13, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 13, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 95% identical to SEQ ID NO: 13, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 13, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 99% identical to SEQ ID NO: 13, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 13, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may comprise the amino acid sequence of SEQ ID NO: 13, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may consist of the amino acid sequence of SEQ ID NO: 13, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 13.

[0242] A Pvs230 or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 13. A Pvs230 or immunogenic fragment thereof according to the invention may comprise or consist of the amino acid sequence of SEQ ID NO: 13.

[0243] Similarly, the invention therefore relates to the use of SEQ ID NO: 83 and fragments and variants thereof. The definitions of Pvs230, immunogenic fragments and variants thereof in the preceding paragraphs in the context of SEQ ID NO: 13 apply equally and without reservation to Pvs230 and immunogenic fragments thereof comprising SEQ ID NO: 83 and fragments and variants thereof. By way of non-limiting example, a variant of SEQ ID NO: 83 may differ from SEQ ID NO: 83 by no more than 10 amino acids, such as no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid; e.g. by substitution or deletion, typically substitution, preferably a conservative amino acid substitution.

[0244] A Pvs230 or immunogenic fragment thereof according to the invention may comprise other components, such as a signal peptide and / or label such as those described herein in the context of fusion proteins of the invention.

[0245] A Pvs230 or immunogenic fragment thereof according to the invention may by glycosylated or unglycosylated, preferably unglycosylated. A Pvs230 or immunogenic fragment thereof according to the invention may lack N-glycosylation. A PvCSP or immunogenic fragment thereof according to the invention may lack O-glycosylation. A Pvs230 or immunogenic fragment thereof according to the invention may lack both N-glycosylation and O-glycosylation.

[0246] The invention also relates to fusion proteins comprising a Pvs230 or immunogenic fragment thereof according to the invention, as well as to VLPs comprising Pvs230 or immunogenic fragment thereof according to the invention. Such fusion proteins and VLPs are described further herein. For the avoidance of doubt, any and all disclosure herein in relation to Pvs230 and immunogenic fragment thereof according to the invention applies equally and unreservedly to other aspects of the invention, such as fusion proteins and VLPs comprising a Pvs230 or immunogenic fragment thereof according to the invention.

[0247] Plasmodium vivax Duffy binding protein II (PvDBPII)

[0248] Plasmodium vivax Duffy binding protein II (PvDBPII) is a key protein domain involved in the invasion of human red blood cells by the malaria parasite Plasmodium vivax. PvDBPII is the receptorbinding region of the Duffy Binding Protein (DBP), a surface protein expressed by P. vivax merozoites. This protein specifically binds to the Duffy Antigen Receptor for Chemokines (DARC) on the surface of human erythrocytes.

[0249] The full-length PvDBPII has an amino acid sequence according to NCBI Accession No. XP_001608387 (version 1, updated 17 April 2018, accessed 17 July 2025). This sequence is herein incorporated by reference in its entirety. The invention relates to the use of fragments of PvDBPII. In particular, the invention relates to the use of fragments of PvDBPII comprising or consisting of amino acid residues 194 to 521 of the PvDBPII amino acid sequence according to NCBI Accession No. XP_001608387 as described herein, or variants thereof. The amino acid sequence of residues 194 to 521 of the PvDBPII amino acid sequence according to NCBI Accession No. XP 001608387 is SEQ ID NO: 92 herein. An exemplary nucleic acid sequence encoding SEQ ID NO: 92 is provided by SEQ ID NO: 118.

[0250] The invention therefore relates to the use of SEQ ID NO: 92 and fragments and variants thereof. A variant of SEQ ID NO: 92 may differ from SEQ ID NO: 92 by no more than 10 amino acids, such as no more than 9, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 amino acid; e.g. by substitution or deletion, typically substitution, preferably a conservative amino acid substitution.

[0251] A PvDBPII or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to SEQ ID NO: 92, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 85% identical to SEQ ID NO: 92, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 92, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 90% identical to SEQ ID NO: 92, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 92, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 95% identical to SEQ ID NO: 92, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 92, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 99% identical to SEQ ID NO: 92, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 92, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may comprise the amino acid sequence of SEQ ID NO: 92, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may consist of the amino acid sequence of SEQ ID NO: 92, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 92.

[0252] A PvDBPII or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 92. A PvDBPII or immunogenic fragment thereof according to the invention may comprise or consist of the amino acid sequence of SEQ ID NO: 92.

[0253] The invention therefore relates to the use of variants of SEQ ID NO: 92 and fragments thereof. A variant of SEQ ID NO: 92 may comprise SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to SEQ ID NO: 93, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 85% identical to SEQ ID NO: 93, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 93, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 90% identical to SEQ ID NO: 93, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 93, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 95% identical to SEQ ID NO: 93, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 93, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may comprise an amino acid sequence that is at least 99% identical to SEQ ID NO: 93, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 93, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may comprise the amino acid sequence of SEQ ID NO: 93, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may consist of the amino acid sequence of SEQ ID NO: 93, or a fragment thereof which is at least 50% of the length of SEQ ID NO: 93.

[0254] A PvDBPII or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 85% identical to SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 95% identical to SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may comprise or consist of an amino acid sequence that is at least 99% identical to SEQ ID NO: 93. A PvDBPII or immunogenic fragment thereof according to the invention may comprise or consist of the amino acid sequence of SEQ ID NO: 93.

[0255] A PvDBPII or immunogenic fragment thereof according to the invention may comprise other components, such as a signal peptide, linker, and / or label such as those described herein in the context of fusion proteins of the invention.

[0256] A PvDBPII or immunogenic fragment thereof according to the invention may by glycosylated or unglycosylated, preferably unglycosylated. A PvDBPII or immunogenic fragment thereof according to the invention may lack N-glycosylation. A PvDBPII or immunogenic fragment thereof according to the invention may lack O-glycosylation. A PvDBPII or immunogenic fragment thereof according to the invention may lack both N-glycosylation and O-glycosylation.

[0257] The invention also relates to fusion proteins comprising a PvDBPII or immunogenic fragment thereof according to the invention, as well as to VLPs comprising PvDBPII or immunogenic fragment thereof according to the invention. Such fusion proteins and VLPs are described further herein. For the avoidance of doubt, any and all disclosure herein in relation to PvDBPII and immunogenic fragment thereof according to the invention applies equally and unreservedly to other aspects of the invention, such as fusion proteins and VLPs comprising a PvDBPII or immunogenic fragment thereof according to the invention.

[0258] Fusion proteins

[0259] The invention also provides a fusion protein. The terms "fusion protein" and "fusion molecule" are used interchangeably and may refer to a polypeptide comprising a first immunogenic polypeptide (antigen or fragment thereof) linked to a second immunogenic polypeptide (antigen or fragment thereof).

[0260] According to the invention, the first immunogenic polypeptide is typically a PvCSP or immunogenic fragment thereof according to the invention and the second immunogenic polypeptide is typically a Pvs230 or immunogenic fragment thereof according to the invention. Thus, the invention provides fusion proteins comprising a PvCSP or immunogenic fragment thereof according to the invention and / or a Pvs230 or immunogenic fragment thereof according to the invention.

[0261] For the avoidance of doubt, a fusion protein of the invention may comprise any PvCSP or immunogenic fragment thereof according to the invention. Similarly, for the avoidance of doubt, a fusion protein of the invention may comprise any Pvs230 or immunogenic fragment thereof according to the invention. The PvCSP or immunogenic fragment thereof according to the invention and the Pvs230 or immunogenic fragment thereof according to the invention may be selected independently.

[0262] Thus, and by way of non-limiting example, a fusion protein of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 85% identical to SEQ ID NO: 3. By way of a further non-limiting example, a fusion protein of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 90% identical to SEQ ID NO: 3. By way of a further non-limiting example, a fusion protein of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 95% identical to SEQ ID NO: 3. By way of a further non-limiting example, a fusion protein of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 99% identical to SEQ ID NO: 3. By way of a further non-limiting example, a fusion protein of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 3. These PvCSP and immunogenic fragments thereof may be used in a fusion protein of the invention in combination with any Pvs230 as described herein.

[0263] Similarly, and by way of non-limiting example, a fusion protein of the invention may comprise a Pvs230 or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 85% identical to SEQ ID NO: 13. By way of a further non-limiting example, a fusion protein of the invention may comprise a Pvs230 or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 90% identical to SEQ ID NO: 13. By way of a further non-limiting example, a fusion protein of the invention may comprise a Pvs230 or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 95% identical to SEQ ID NO: 13. By way of a further non-limiting example, a fusion protein of the invention may comprise a Pvs230 or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 99% identical to SEQ ID NO: 13. By way of a further non-limiting example, a fusion protein of the invention may comprise a Pvs230 or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 13. These Pvs230 and immunogenic fragments thereof may be used in a fusion protein of the invention in combination with any PvCSP as described herein, particularly any chimeric PvCSP described herein. The polypeptide sequence of the first immunogenic polypeptide (e.g. comprising a PvCSP or immunogenic fragment thereof according to the invention) and the second immunogenic polypeptide (e.g. a Pvs230 or immunogenic fragment thereof according to the invention) may be covalently linked (e.g., fused) by recombinant, chemical or other suitable methods. The fusion protein can be fused at one or several sites through a linker, particularly a peptide linker sequence, examples of which are described herein. Alternatively, the peptide linker may be used to assist in construction of the fusion protein.

[0264] The immunogenic polypeptides (e.g., the PvCSP or immunogenic fragment thereof and the Pvs230 or immunogenic fragment thereof according to the invention) may be directly fused. Alternatively, the fusion protein may comprise a linker between the immunogenic polypeptides (e.g., the PvCSP or immunogenic fragment thereof and the Pvs230 or immunogenic fragment thereof according to the invention). Wherein a linker is present, preferably the linker is a flexible linker (e.g., allowing effective positioning of the two immunogenic polypeptides). The flexible linker may, for example, be a peptide linker. The peptide linker may comprise (or consist of) 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 amino acids. The peptide linker may comprise (or consist of) from 10 to 20 amino acids; preferably from 12 to 18 amino acids. Preferably, the immunogenic polypeptides (e.g., the PvCSP or immunogenic fragment thereof and the Pvs230 or immunogenic fragment thereof according to the invention) are directly fused, such that the C-terminal residue of the N-terminal domain is linked directly (e.g. via a peptide bond) to the N-terminal residue of the C-terminal domain.

[0265] Different orientations of the immunogenic polypeptides (e.g., the PvCSP or immunogenic fragment thereof and the Pvs230 or immunogenic fragment thereof according to the invention) are envisaged. Thus, according to the invention, a fusion protein may comprise the PvCSP protein or immunogenic fragment thereof N-terminal to Pvs230 protein or the immunogenic fragment thereof. Alternatively, the PvCSP protein or immunogenic fragment thereof may be C-terminal to Pvs230 protein or the immunogenic fragment thereof. Preferably, the PvCSP protein or immunogenic fragment thereof is N-terminal to the Pvs230 protein or the immunogenic fragment thereof.

[0266] The immunogenic polypeptides (e.g., the PvCSP or immunogenic fragment thereof, the Pvs230 or immunogenic fragment thereof and the PvDBPII or immunogenic fragment thereof according to the invention) may be directly fused. Alternatively, the fusion protein may comprise a linker between the immunogenic polypeptides (e.g., the PvCSP or immunogenic fragment thereof, the Pvs230 or immunogenic fragment thereof and / or the PvDBPII or immunogenic fragment thereof according to the invention). Wherein a linker is present, preferably the linker is a flexible linker (e.g., allowing effective positioning of the two immunogenic polypeptides). The flexible linker may, for example, be a peptide linker. The peptide linker may comprise (or consist of) 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 amino acids. The peptide linker may comprise (or consist of) from 1 to 12 amino acids; preferably from 3 to 12 amino acids. Optionally, the immunogenic polypeptides (e.g., the PvCSP or immunogenic fragment thereof, the Pvs230 or immunogenic fragment thereof and / or the PvDBPII or immunogenic fragment thereof according to the invention) are directly fused, such that the C-terminal residue of the N-terminal domain is linked directly (e.g. via a peptide bond) to the N-terminal residue of the C-terminal domain.

[0267] Different orientations of the immunogenic polypeptides (e.g., the PvCSP or immunogenic fragment thereof, the Pvs230 or immunogenic fragment thereof and the PvDBPII or immunogenic fragment thereof according to the invention) are envisaged. Thus, according to the invention, a fusion protein may comprise the PvDBPII protein or immunogenic fragment thereof N-terminal to Pvs230 protein or the immunogenic fragment thereof. Alternatively, the PvDBPII protein or immunogenic fragment thereof may be C-terminal to Pvs230 protein or the immunogenic fragment thereof. According to the invention, a fusion protein may comprise the PvDBPII protein or immunogenic fragment thereof N-terminal to PvCSP protein or the immunogenic fragment thereof. Alternatively, the PvDBPII protein or immunogenic fragment thereof may be C-terminal to PvCSP protein or the immunogenic fragment thereof. Further, according to the invention a fusion protein may comprise: (i) the PvCSP or immunogenic fragment thereof N-terminal to the PvDBPII protein or immunogenic fragment thereof which is itself N-terminal to the Pvs230 or immunogenic fragment thereof (PvCSP-PvDBPII-Pvs230); (ii) the PvCSP or immunogenic fragment thereof N-terminal to the Pvs230 protein or immunogenic fragment thereof which is itself N-terminal to the PvDBPII or immunogenic fragment thereof (PvCSP-Pvs230-PvDBPII); (iii) the PvDBPII or immunogenic fragment thereof N-terminal to the Pvs230 protein or immunogenic fragment thereof which is itself N-terminal to the PvCSP or immunogenic fragment thereof (PvDBPII-Pvs230-PvCSP); (iv) the PvDBPII or immunogenic fragment thereof N-terminal to the PvCSP protein or immunogenic fragment thereof which is itself N-terminal to the Pvs230 or immunogenic fragment thereof (PvDBPII-PvCSP-Pvs230); (v) the Pvs230 or immunogenic fragment thereof N-terminal to the PvCSP protein or immunogenic fragment thereof which is itself N-terminal to the PvDBPII or immunogenic fragment thereof (Pvs230-PvCSP-PvDBPII); or (v) the Pvs230 or immunogenic fragment thereof N-terminal to the PvDBPII protein or immunogenic fragment thereof which is itself N-terminal to the PvCSP or immunogenic fragment thereof (Pvs230-PvDBPII-PvCSP). Preferably, the PvCSP or immunogenic fragment thereof is: (i) N-terminal to the PvDBPII protein or immunogenic fragment thereof, which is itself N-terminal to the Pvs230 or immunogenic fragment thereof (PvCSP-PvDBPII-Pvs230); or (ii) the PvCSP or immunogenic fragment thereof is N-terminal to the Pvs230 protein or immunogenic fragment thereof which is itself N-terminal to the PvDBPII or immunogenic fragment thereof (PvCSP-Pvs230-PvDBPII).

[0268] The fusion protein may further comprise a signal peptide. The signal peptide may be located at the N-or C-terminus of the fusion protein. Preferably, the signal peptide may be located at the N-terminus of the fusion protein. The signal peptide may comprise an amino acid sequence selected from the group consisting of: SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16. The signal peptide may consist of an amino acid sequence selected from the group consisting of: SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16. A fusion protein of the invention may further comprise a first complexing motif. Said first complexing motif may be located at the N-, C-terminus or positioned between two of the PvCSP or immunogenic fragment thereof, the Pvs230 or immunogenic fragment thereof and the PvDBPII or immunogenic fragment thereof of the fusion protein. Preferably, the first complexing motif may be located at the C-terminus or positioned between two of the PvCSP or immunogenic fragment thereof, the Pvs230 or immunogenic fragment thereof and the PvDBPII or immunogenic fragmentthereof of the fusion protein. More preferably, the first complexing motif may be located at the C-terminus of the fusion protein. The first complexing motif may comprise an amino acid sequence selected from the group consisting of: SEQ ID NO: 17 (4T), SEQ ID NO: 32 (Isopeptag-N), SEQ ID NO: 33 (Isopeptag), SEQ ID NO: 34 (Isopeptag), SEQ ID NO: 35 (SdyTag), SEQ ID NO: 36 (SilkTag) and SEQ ID NO: 37 (DogTag). Preferably, the first complexing motif may comprise SEQ ID NO: 17. The first complexing motif may consist of an amino acid sequence selected from the group consisting of: SEQ ID NO: 17, SEQ ID NO:32 (Isopeptag-N), SEQ ID NO:33 (Isopeptag), SEQ ID NO:34 (Isopeptag), SEQ ID NO:35 (SdyTag), SEQ ID NO:36 (SilkTag) and SEQ ID NO: 37 (DogTag). Preferably, the first complexing motif may consist of SEQ ID NO: 17.

[0269] The first complexing motif may be separated from the remainder of the fusion protein by a linker. Said linker may be a linker polypeptide, typically a flexible linker polypeptide. The linker polypeptide may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 amino acid residues in length. The linker polypeptide may comprise the amino acid sequence of SEQ ID NO: 18. Alternatively or in addition, the linker polypeptide may comprise the amino acid sequence of SEQ ID NO: 19. Alternatively or in addition, the linker polypeptide may comprise the amino acid sequence of SEQ ID NO: 67. Alternatively or in addition, the linker polypeptide may comprise the amino acid sequence of SEQ ID NO: 68. Alternatively or in addition, the linker polypeptide may comprise the amino acid sequence of SEQ ID NO: 100. Alternatively or in addition, the linker polypeptide may comprise the amino acid sequence of SEQ ID NO: 101. Alternatively or in addition, the linker polypeptide may comprise the amino acid sequence of SEQ ID NO: 115. A linker peptide may comprise multiple copies (e.g. 2, 3, 4, 5 or more copies) of a linker sequence, such as SEQ ID NO: 18, 19, 67, 68, 100, 101 or 115. The linker peptide may consist of the amino acid sequence of SEQ ID NO: 18. Alternatively or in addition, the linker peptide may consist of the amino acid sequence of SEQ ID NO: 19. Alternatively or in addition, the linker peptide may consist of the amino acid sequence of SEQ ID NO: 67. Alternatively or in addition, the linker peptide may consist of the amino acid sequence of SEQ ID NO: 68. Alternatively or in addition, the linker peptide may consist of the amino acid sequence of SEQ ID NO: 100. Alternatively or in addition, the linker peptide may consist of the amino acid sequence of SEQ ID NO: 101. Alternatively or in addition, the linker peptide may consist of the amino acid sequence of SEQ ID NO: 115. A linker peptide may consist of multiple copies (e.g. 2, 3, 4, 5 or more copies) of a linker sequence, such as multiple copies of SEQ ID NO: 67 (e.g. SEQ ID NO: 18), 68 (e.g. SEQ ID NO: 19) or 100 (e.g. SEQ ID NO: 115). A fusion protein of the invention may further comprise a label. Said label may be located at the N- or C-terminus of the fusion protein. The label may be selected from the group consisting of: histag (SEQ ID NO: 38), GST (SEQ ID NO: 39), MBP (SEQ ID NO: 40), NusA (SEQ ID NO: 41) and SUMO (SEQ ID NO: 42). Preferably, the label may be a histag (SEQ ID NO: 38).

[0270] Accordingly, a fusion protein according to the invention may comprise or consist of an amino acid sequence that is at least 80% identical (e.g. at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or more identical) to any one of SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126. A Fusion protein according to the invention may comprise an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126. A Fusion protein according to the invention may consist of an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126. A Fusion protein according to the invention may comprise an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126. A Fusion protein according to the invention may consist of an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126. A Fusion protein according to the invention may comprise an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126. A Fusion protein according to the invention may consist of an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126. A Fusion protein according to the invention may comprise an amino acid sequence that is at least 99% identical to any one of SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126. A Fusion protein according to the invention may consist of an amino acid sequence that is at least 99% identical to any one of SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126. A Fusion protein according to the invention may comprise the amino acid sequence of any one to any one of SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126. A Fusion protein according to the invention may consist of the amino acid sequence of any one to any one of SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126.

[0271] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 20. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 20. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 20.

[0272] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 21. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 21. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 21.

[0273] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 22. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 22. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 22.

[0274] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 23. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 23. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 23.

[0275] In some preferred embodiment of the invention, a fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 24. In some further preferred embodiment of the invention, a fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 24. In some further preferred embodiment of the invention, a fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 24.

[0276] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 25. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 25. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 25.

[0277] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 26. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 26. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 26.

[0278] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 27. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 27. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 27.

[0279] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 63. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 63. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 63.

[0280] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 64. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 64. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 64.

[0281] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 65. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 65. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 65. A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 66. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 66. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 66.

[0282] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 94. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 94. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 94.

[0283] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 95. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 95. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 95.

[0284] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 96. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 96. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 96.

[0285] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 97. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 97. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 97.

[0286] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 98. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 98. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 98.

[0287] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 99. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 99. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 99.

[0288] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 102. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 102. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 102.

[0289] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 103. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 103. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 103. A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 104. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 104. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 104.

[0290] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 105. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 105. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 105.

[0291] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 106. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 106. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 106.

[0292] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 107. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 107. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 107.

[0293] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 108. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 108. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 108.

[0294] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 109. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 109. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 109.

[0295] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 110. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 110. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 110.

[0296] A fusion protein may comprise or consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 126. A fusion protein may comprise or consist of an amino acid sequence that is at least 90% identical to SEQ ID NO: 126. A fusion protein may comprise or consist of the amino acid sequence of SEQ ID NO: 126.

[0297] A fusion protein according to the invention may by glycosylated or unglycosylated, preferably unglycosylated. A fusion protein according to the invention may lack N-glycosylation. A fusion protein according to the invention may lack O-glycosylation. A fusion protein according to the invention may lack both N-glycosylation and O-glycosylation. A fusion protein may further comprise one or more additional immunogenic polypeptide (antigen or fragment thereof). Said one or more additional immunogenic polypeptide (antigen or fragment thereof) any be an additional P. vivax polypeptide, or a variant, derivative or fragment thereof. Said one or more additional immunogenic polypeptide (antigen or fragment thereof) any be a P. falciparum polypeptide, or a variant, derivative or fragment thereof. The one or more additional P. vivax immunogenic polypeptide, or a variant, derivative or fragment thereof may be selected from PvDBPII or PvRBP2b. The one or more additional P. falciparum immunogenic polypeptide, or a variant, derivative or fragment thereof may be selected from the group consisting of PfCSP, PfRH5, PfAMAl, PfMSPl, PfMSP3, PfEBA-175, PfAMAl-RON2, Pf25, Pf23O and Pf48 / 45.

[0298] Virus-like particles (VLPs)

[0299] Virus-like particles (VLPs) are self-assembling, nanoscale structures that resemble the organization and morphology of viruses but lack the viral genetic material necessary for replication and infection. Composed primarily of one or more viral structural proteins (capsid or envelope proteins), VLPs can spontaneously assemble into shapes resembling the original virus's capsid or envelope, depending on the proteins involved.

[0300] A VLP according to the invention may comprise any PvCSP or immunogenic fragment thereof according to the invention, particularly any chimeric PvCSP or immunogenic fragment thereof according to the invention. Alternatively or in addition, a VLP according to the invention may comprise a Pvs230 or immunogenic fragment thereof according to the invention. Further alternatively or in addition, a VLP according to the invention may comprise a PvDBPII or immunogenic fragment thereof according to the invention. The PvCSP or immunogenic fragment thereof according to the invention, the Pvs230 or immunogenic fragment thereof and the PvDBPII or immunogenic fragment thereof according to the invention may be selected independently. A VLP according to the invention may comprise any fusion protein of the invention. The fusion protein comprised in a VLP according to the invention may comprise any PvCSP or immunogenic fragment thereof according to the invention and / or any Pvs230 or immunogenic fragment thereof according to the invention and / or any PvDBPII or immunogenic fragment thereof according to the invention. The PvCSP or immunogenic fragment thereof according to the invention, the Pvs230 or immunogenic fragment thereof according to the invention and the PvDBPII or immunogenic fragment thereof according to the invention comprised in said fusion protein may be selected independently.

[0301] Thus, and by way of non-limiting example, a VLP of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 85% identical to SEQ ID NO: 3. By way of a further non-limiting example, a VLP of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 90% identical to SEQ ID NO: 3. By way of a further non-limiting example, a VLP of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 95% identical to SEQ ID NO: 3. By way of a further non-limiting example, a VLP of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 99% identical to SEQ ID NO: 3. By way of a further non-limiting example, a VLP of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 3.

[0302] Thus, and by way of non-limiting example, a VLP of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 85% identical to SEQ ID NO: 91. Byway of a further non-limiting example, a VLP of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 90% identical to SEQ ID NO: 91. By way of a further non-limiting example, a VLP of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 95% identical to SEQ ID NO: 91. By way of a further non-limiting example, a VLP of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 99% identical to SEQ ID NO: 91. By way of a further non-limiting example, a VLP of the invention may comprise a PvCSP or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 91.

[0303] Alternatively or in addition, and by way of further non-limiting example, a VLP of the invention may comprise a Pvs230 or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 85% identical to SEQ ID NO: 13. By way of a further non-limiting example, a VLP of the invention may comprise a Pvs230 or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 90% identical to SEQ ID NO: 13. By way of a further non-limiting example, a VLP of the invention may comprise a Pvs230 or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 95% identical to SEQ ID NO: 13. By way of a further non-limiting example, a VLP of the invention may comprise a Pvs230 or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 99% identical to SEQ ID NO: 13. By way of a further non-limiting example, a VLP of the invention may comprise a Pvs230 or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 13.

[0304] Similarly, and by way of further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 20, 21, 22, 23, 24, 25, 26, 27, 63, 64, 65, 66, 94, 95, 96, 97, 98, 99, 102, 103, 104, 105, 106, 107, 108, 109, 110 and 126. By way of a further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 20, 21, 22, 23, 24, 25, 26, 27, 63, 64, 65, 66, 94, 95, 96, 97, 98, 99, 102, 103, 104, 105, 106, 107, 108, 109, 110 and 126. By way of a further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 20, 21, 22, 23, 24, 25, 26, 27, 63, 64, 65, 66,94, 95, 96, 97, 98, 99, 102, 103, 104, 105, 106, 107, 108, 109, 110 and 126. By way of a further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 99% identical to any one of SEQ ID NOs: 20, 21, 22, 23, 24, 25, 26, 27, 63, 64, 65, 66, 94, 95, 96, 97, 98, 99, 102, 103, 104, 105, 106, 107, 108, 109, 110 and 126. By way of a further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 20, 21, 22, 23, 24, 25, 26, 27, 63, 64, 65, 66, 94, 95, 96, 97, 98, 99, 102, 103, 104, 105, 106, 107, 108, 109, 110 and 126.

[0305] By way of further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 20, 21, 22, 23, 24, 94, 99, 102, 103, 104, 105, 106 and 107. By way of a further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 20, 21, 22, 23, 24, 94, 99, 102, 103, 104, 105, 106 and 107. By way of a further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 20, 21, 22, 23, 24, 94, 99, 102, 103, 104, 105, 106 and 107. By way of a further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 99% identical to any one of SEQ ID NOs: 20, 21, 22, 23, 24, 94, 99, 102, 103, 104, 105, 106 and 107. By way of a further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 20, 21, 22, 23, 24, 94, 99, 102, 103, 104, 105, 106 and 107.

[0306] By way of further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 85% identical to any one of SEQ ID NOs: 20, 24, 94, 99, 102, 103, 104, 105, 106 and 107. By way of a further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 90% identical to any one of SEQ ID NOs: 20, 24, 94, 99, 102, 103, 104, 105, 106 and 107. By way of a further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 95% identical to any one of SEQ ID NOs: 20, 24, 94, 99, 102, 103, 104, 105, 106 and 107. By way of a further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 99% identical to any one of SEQ ID NOs: 20, 24, 94, 99, 102, 103, 104, 105, 106 and 107. By way of a further non-limiting example, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 20, 24, 94, 99, 102, 103, 104, 105, 106 and 107.

[0307] In some preferred embodiment of the invention, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 24. In some further preferred embodiment of the invention, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 90% identical to SEQ ID NO: 24. In some further preferred embodiment of the invention, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 24.

[0308] In some preferred embodiment of the invention, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 105. In some further preferred embodiment of the invention, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 90% identical to SEQ ID NO: 105. In some further preferred embodiment of the invention, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 105.

[0309] In some preferred embodiment of the invention, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 107. In some further preferred embodiment of the invention, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of an amino acid sequence that is at least 90% identical to SEQ ID NO: 107. In some further preferred embodiment of the invention, a VLP of the invention may comprise a fusion protein or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 107.

[0310] A VLP according to the invention may comprise a core comprising or consisting of at least one subunit of a VLP carrier. As used herein "a subunit of a VLP carrier" may refer to a capsid protein or envelope protein of a virus from which a VLP is derived. The at least one subunit of a VLP carrier may be selected from the group consisting of the amino acid sequence of: SEQ ID NO: 28, Hepatitis B surface antigen (SEQ ID NO: 43), HPV 18 LI protein (SEQ ID NO: 44), Hepatitis E P239 protein (SEQ ID NO: 45), or HPV 16 LI protein (SEQ ID NO: 46), AP205 (SEQ ID NO: 47), Ferritin (SEQ ID NO: 48), I53-50A (SEQ ID NO: 49) or I53-50B (SEQ ID NO: 69), or a sequence which has at least 80% (e.g. at least 85%, at least 90%, at least 95%, at least 99% or more identity) to any one of SEQ ID NOs: 43 to 49 or 69. Preferably the at least one subunit of a VLP carrier comprises or consists of an amino acid sequence SEQ ID NO: 28, or a sequence which has at least 80% (e.g. at least 85%, at least 90%, at least 95%, at least 99% or more identity) to SEQ ID NO: 28. Particularly preferably, the at least one subunit of a VLP carrier comprises or consists of an amino acid sequence SEQ ID NO: 28.

[0311] A VLP of the invention may comprise at least 2, at least 3, at least 4, at least 12, at least 14, at least 24, at least 60 or more subunits of a VLP carrier. These may be independently selected from any suitable VLP carrier subunits known in the art, such as those described herein. Thus, a VLP of the invention may comprise at least one copy of each of two different carrier subunits. Typically, the VLP carrier subunits of a VLP of the invention are identical.

[0312] A PvCSP or immunogenic fragment thereof according to the invention, a Pvs230 or immunogenic fragment thereof according to the invention, a PvDBPII or immunogenic fragment thereof and / or a fusion protein according to the invention may be linked to at least one subunit of a VLP carrier. The PvCSP or immunogenic fragment thereof according to the invention, the Pvs230 or immunogenic fragment thereof according to the invention, the PvDBPII or immunogenic fragment thereof and / or fusion protein according to the invention may be directly or indirectly linked to at least one subunit of a VLP carrier. A direct link may be a covalent link by recombinant, chemical or other suitable methods. An indirect link may be via a linker, such as a flexible linker, which may form a covalent link between the VLP carrier subunit and the PvCSP or immunogenic fragments thereof, Pvs230 or immunogenic fragments thereof, PvDBPII or immunogenic fragment thereof or fusion protein. Examples of linkers, particularly peptide linkers are described herein.

[0313] A PvCSP or immunogenic fragment thereof according to the invention, a Pvs230 or immunogenic fragment thereof according to the invention or a PvDBPII or immunogenic fragment thereof according to the invention may be selected independently and / or a fusion protein according to the invention have a first complexing motif attached at the N- and / or C-terminus, preferably the C-terminus. First complexing motifs are described above in the context of fusion proteins of the invention. This disclosure applies equally and without reservation to PvCSP or immunogenic fragments thereof according to the invention, Pvs230 or immunogenic fragments thereof according to the invention and PvDBPII or immunogenic fragment thereof according to the invention, particularly when said PvCSP or immunogenic fragments thereof, Pvs230 or immunogenic fragments thereof or PvDBPII or immunogenic fragment thereof are present in VLPs of the invention. Thus, by way of non-limiting example, a first complexing motif may be any one of SEQ ID NOs: 17 and 32 to 37, particularly SEQ ID NO: 17. By way of further nonlimiting example, said first complexing motif may be linked to a PvCSP or immunogenic fragment, a Pvs230 or immunogenic fragment thereof, a PvDBPII or immunogenic fragment thereof or fusion protein directly or indirectly, such as by a linker (e.g. those exemplified herein, such as SEQ ID NO: 18). Linkage of the PvCSP or immunogenic fragments thereof, Pvs230 or immunogenic fragments thereof, PvDBPII or immunogenic fragment thereof or fusion protein of the invention to at least one subunit of the VLP carrier typically presents the PvCSP or immunogenic fragments thereof, Pvs230 or immunogenic fragments thereof, PvDBPII or immunogenic fragment thereof or fusion protein on the surface of the VLP. This surface presentation means that the PvCSP or immunogenic fragments thereof, Pvs230 or immunogenic fragments thereof, PvDBPII or immunogenic fragment thereof or fusion protein is exposed to a subject's immune system on administration of the PvCSP or immunogenic fragments thereof, Pvs230 or immunogenic fragments thereof, PvDBPII or immunogenic fragment thereof or fusion protein to the subject, enabling an immune response to be elicited.

[0314] The at least one subunit of the VLP carrier may further comprise a second complexing motif. This second complexing motif typically interacts with a first complexing motif on the PvCSP or immunogenic fragment thereof, Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof or fusion protein of the invention to link the PvCSP or immunogenic fragment thereof, Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof or fusion protein to the at least one subunit of the VLP carrier. The second complexing motif may be present at the N-terminus of the at least one VLP carrier subunit. The second complexing motif may be present at the C-terminus of the at least one VLP carrier subunit. Preferably, the second complexing motif is present at the N-terminus of the at least one VLP carrier subunit.

[0315] The second complexing motif may comprise or consist of an amino acid sequence selected from the group consisting of: SEQ ID NO: 29, pilin-N (SEQ ID NO: 50), pilin-C (SEQ ID NO: 51), SnoopCatcher (SEQ ID NO: 52), SdyCatcher (SEQ ID NO: 53), SilkCatcher (SEQ ID NO: 54) or DogCatcher (SEQ ID NO: 55). Preferably, the second complexing motif comprises or consists of SEQ ID NO: 29.

[0316] The second complexing motif may be separated from the remainder of the VLP carrier subunit by a linker polypeptide, which may be flexible. Flexible linker polypeptides are known in the art and selection of such a linker is within the routine practice of one of ordinary skill in the art. By way of non-limiting examples, linker peptides may comprise one or more copies (e.g. 2, 3, 4, 5, 6, 7, 8, 9, 10 or more copies) of GGGGS (SEQ ID NO: 67), EAAAK (SEQ ID NO: 68), GSG (SEQ ID NO: 100), GGGS (SEQ ID NO: 101) or G. The linker polypeptide may comprise or consist of an amino acid sequence selected SEQ ID NO: 18, 19, 67, 68, 100, 101 or 115. The linker polypeptide may comprise or consist of the amino acid sequence of SEQ ID NO: 18. The linker polypeptide may comprise or consist of the amino acid sequence of SEQ ID NO: 19. Alternatively or in addition, the linker polypeptide may comprise or consist of the amino acid sequence of SEQ ID NO: 67. Alternatively or in addition, the linker polypeptide may comprise or consist of the amino acid sequence of SEQ ID NO: 68. Alternatively or in addition, the linker polypeptide may comprise or consist of the amino acid sequence of SEQ ID NO: 100. Alternatively or in addition, the linker polypeptide may comprise or consist of the amino acid sequence of SEQ ID NO: 101. Alternatively or in addition, the linker polypeptide may comprise or consist of the amino acid sequence of SEQ ID NO: 115.

[0317] The VLP core may comprise an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent. The VLP core may consist of an amino acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent. The VLP core may comprise the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent. The VLP core may consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent. The VLP core of SEQ ID NO: 30 is described in WO 2023 / 109195, which is herein incorporated by reference in its entirety. In particular Example 4 of WO 2023 / 109195 describes a combined preparation process for the production of this VLP core using urea and NaCI. Example 5 of WO 2023 / 109195 describes a two-step chromatographic purification process for said VLP core.

[0318] Typically, the first complexing motif and the second complexing motif are covalently linked, such that at least one PvCSP or immunogenic fragment thereof, at least one Pvs230 or immunogenic fragment thereof, at least one PvDBPII or immunogenic fragment thereof and / or at least one fusion protein of the invention is presented on the surface of the VLP.

[0319] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a fusion protein which comprises or consists of the amino acid sequence of SEQ ID NO: 20, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the fusion protein comprises or consists of the amino acid sequence of SEQ ID NO: 20.

[0320] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a fusion protein which comprises or consists of the amino acid sequence of SEQ ID NO: 21, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the fusion protein comprises or consists of the amino acid sequence of SEQ ID NO: 21.

[0321] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a fusion protein which comprises or consists of the amino acid sequence of SEQ ID NO: 22, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the fusion protein comprises or consists of the amino acid sequence of SEQ ID NO: 22.

[0322] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a fusion protein which comprises or consists of the amino acid sequence of SEQ ID NO: 23, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the fusion protein comprises or consists of the amino acid sequence of SEQ ID NO: 23.

[0323] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a fusion protein which comprises or consists of the amino acid sequence of SEQ ID NO: 24, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the fusion protein comprises or consists of the amino acid sequence of SEQ ID NO: 24.

[0324] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a fusion protein which comprises or consists of the amino acid sequence of SEQ ID NO: 25, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the fusion protein comprises or consists of the amino acid sequence of SEQ ID NO: 25.

[0325] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a fusion protein which comprises or consists of the amino acid sequence of SEQ ID NO: 26, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the fusion protein comprises or consists of the amino acid sequence of SEQ ID NO: 26.

[0326] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a fusion protein which comprises or consists of the amino acid sequence of SEQ ID NO: 27, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the fusion protein comprises or consists of the amino acid sequence of SEQ ID NO: 27.

[0327] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a fusion protein which comprises or consists of the amino acid sequence of SEQ ID NO: 63, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the fusion protein comprises or consists of the amino acid sequence of SEQ ID NO: 63.

[0328] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a fusion protein which comprises or consists of the amino acid sequence of SEQ ID NO: 64, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the fusion protein comprises or consists of the amino acid sequence of SEQ ID NO: 64.

[0329] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a fusion protein which comprises or consists of the amino acid sequence of SEQ ID NO: 65, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the fusion protein comprises or consists of the amino acid sequence of SEQ ID NO: 65.

[0330] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a fusion protein which comprises or consists of the amino acid sequence of SEQ ID NO: 66, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the fusion protein comprises or consists of the amino acid sequence of SEQ ID NO: 66.

[0331] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a PvCSP or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 3, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the PvCSP or immunogenic fragment comprises or consists of the amino acid sequence of SEQ ID NO: 3.

[0332] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a PvCSP or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 56, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the PvCSP or immunogenic fragment comprises or consists of the amino acid sequence of SEQ ID NO: 56.

[0333] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a PvCSP or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 57, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the PvCSP or immunogenic fragment comprises or consists of the amino acid sequence of SEQ ID NO: 57.

[0334] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a PvCSP or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 58, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the PvCSP or immunogenic fragment comprises or consists of the amino acid sequence of SEQ ID NO: 58.

[0335] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a PvCSP or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 59, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the PvCSP or immunogenic fragment comprises or consists of the amino acid sequence of SEQ ID NO: 59.

[0336] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a PvCSP or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 60, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the PvCSP or immunogenic fragment comprises or consists of the amino acid sequence of SEQ ID NO: 60.

[0337] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a PvCSP or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 61, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the PvCSP or immunogenic fragment comprises or consists of the amino acid sequence of SEQ ID NO: 61.

[0338] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a PvCSP or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 62, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the PvCSP or immunogenic fragment comprises or consists of the amino acid sequence of SEQ ID NO: 62.

[0339] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a PvCSP or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 84, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the PvCSP or immunogenic fragment comprises or consists of the amino acid sequence of SEQ ID NO: 84.

[0340] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a Pvs230 or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 13, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the Pvs230 or immunogenic fragment comprises or consists of the amino acid sequence of SEQ ID NO: 13.

[0341] The invention provides a VLP in which (i) the VLP core comprises or consists of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity); and (ii) a Pvs230 or immunogenic fragment thereof which comprises or consists of the amino acid sequence of SEQ ID NO: 83, or an amino acid sequence with at least 80% identity thereto as described herein (e.g. at least 85%, 90%, 95% or 99% identity). In particular, the invention provides a VLP in which (i) the VLP core comprises or consist of the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent; and (ii) the Pvs230 or immunogenic fragment comprises or consists of the amino acid sequence of SEQ ID NO: 83.

[0342] The ratio of the VLP core (at least one VLP carrier subunit) to the PvCSP protein or immunogenic fragment thereof, Pvs230 protein or immunogenic fragment thereof, or fusion protein is not particularly limited (i.e. the VLP:antigen ratio). It is within the routine practice of one of ordinary skill in the art to determine an appropriate VLP:antigen ratio, such as using the methods exemplified herein. Typically, the VLP:antigen ratio is selected to maximise the presentation of the PvCSP protein or immunogenic fragment thereof, Pvs230 protein or immunogenic fragment thereof, or fusion protein on the surface of the VLP core. By way of non-limiting example, a VLP:antigen ratio of between about 1:1 to about 1:20, such as between about 1:1 to about 1:10 may be used. The VLP:antigen ratio may be about 1:5, about 1:6, about 1:7, about 1:8, about 1:9 or about 1:10. The VLP:antigen ratio may be about 1:6.

[0343] The diameter of a VLP of the invention less than about 100 nm, such as less than about 95 nm, less than about 90 nm, less than about 85 nm, less than about 80 nm, less than about 75 nm, less than about 70 nm, less than about 60 nm, less than about 55 nm, less than about 50 nm, or less. Typically, a VLP of the invention may have a dimeter of less than about 75 nm, preferably less than about 55nm.

[0344] A VLP of the invention may have a diameter of between about 1 nm to about 90 nm, such as between about 1 nm to about 80 nm, between about 1 nm to about 75 nm, between about 10 nm to about 90 nm, between about 10 nm to about 75 nm, between about 10 nm to about 55 nm, between about 20 nm to about 90 nm, between about 20 nm to about 75 nm, between about 20 nm to about 55 nm, between about 30 nm to about 90 nm, between about 30 nm to about 75 nm, or between about 30 nm to about 55 nm. Preferably, a VLP of the invention may have a diameter of between about 30 nm to about 55 nm. Diameter may be measured for a single VLP, or as an average across a population of VLPs. The diameter of the VLPs of the invention can be measured by methods known by the skilled person, for example dynamic light scattering (DLS) or electron microscopy. Non-limiting examples of suitable DLS protocols are described in the Examples herein.

[0345] Nucleic acids

[0346] The invention provides a nucleic acid encoding any PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof, fusion protein or VLP of the invention. Said nucleic acid may be an isolated nucleic acid molecule. Any reference herein to a nucleic acid or nucleic acid molecule applies equally and without reservation to an isolated nucleic acid molecule.

[0347] A nucleic acid molecule of the invention may comprise or consist of a nucleic acid sequence that is at least 80%, at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 31, 74-82 or 116-125, preferably SEQ ID NO: 31, 120 or 125. A nucleic acid of the invention may comprise or consist of the nucleic acid sequence SEQ ID NO: 31, 74-82 or 116-125, preferably SEQ ID NO: 31, 120 or 125.

[0348] A nucleic acid of the invention acid encoding any PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof or fusion protein of the invention may further comprise or consist of a nucleic acid sequence encoding a VLP core. By way of example, a nucleic acid may further comprise or consist of a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent.

[0349] A nucleic acid encoding a specific protein may be codon optimised for a specific expression cell type. The nucleic acid may comprise any codon specifically encoding the amino acid of the target protein. The skilled person would understand that redundant codons encodes for the same amino acid and are therefore equivalent for the purpose of the invention.

[0350] Expression systems and host cells

[0351] A PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, fusion protein or VLP of the invention may be expressed using any suitable systems.

[0352] A PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, fusion protein or VLP of the invention of the invention may be expressed using conventional systems. Such a system may be a prokaryotic or a eukaryotic system. Examples of such systems are well-known in the art. Non-limiting examples of suitable host systems include Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris, non-lytic insect cell expression systems such as Schneider 2 (S2) and Schneider 3 (S3) cells from Drosophila melanogaster and Sf9 and Sf21 cells from Spodoptera frugiperda, and mammalian expression systems such as CHO cells and human embryonic kidney (HEK / HEK293) cells.

[0353] A host cell may comprise a recombinant expression vector which encodes for a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, fusion protein or VLP of the invention. The host cell is an insect cell, preferably a Drosophila melanogaster cell, or a Pichia yeast cell, or an Escherichia coli cell. The host cell may be a mammalian cell, such as a CHO cell.

[0354] According to the invention it is provided a host cell comprising a nucleic acid or DNA plasmid of the invention. The host cell may be a mammalian cell, a yeast cell or a bacterial cell. The host cell may be transfected with a nucleic acid or DNA plasmid of the invention or may be engineered to express the protein encoded by a nucleic acid or DNA plasmid of the invention. The PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, fusion protein or VLP of the invention may be expressed by a single vector and / or host cell. The PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, fusion protein or VLP of the invention may alternatively be expressed by multiple vectors and / or host cells. By way of non-limiting example, a first vector and / or host cell may be used to express a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, or fusion protein of the invention; and a second vector and / or host cell may be used to express a VLP core of the invention.

[0355] Viral vectors, RNA vectors and DNA plasmids

[0356] The present invention provides a vector encoding a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein or a VLP of the invention.

[0357] The present invention also provides two or more vectors, typically a pair of vectors encoding a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein or a VLP of the invention. In particular, the present invention provides a pair of vectors encoding a VLP of the invention. The first vector may encode a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, or a fusion protein of the invention. The second vector may encode a VLP core of the invention (e.g. at least one VLP carrier subunit) of the invention.

[0358] The vector (vectors) may be present in the form of a formulation or composition.

[0359] The vector(s) may be a viral vector. Such a viral vector may be an adenoviral vector, or a modified Vaccinia Ankara viral vector, a Vesicular stomatitis virus (VSV) vector, an Adeno-associated virus (AAV) vector, a Measles virus vector, a Retrovirus vector, a Herpesvirus vector, or an Influenza virus vector. Preferably, a viral vector may be an adenoviral vector or a modified Vaccinia Ankara viral vector.

[0360] Viral vectors are usually non-replicating or replication impaired vectors, which means that the viral vector cannot replicate to any significant extent in normal cells (e.g. normal human cells), as measured by conventional means - e.g. via measuring DNA synthesis and / or viral titre. Non-replicating or replication impaired vectors may have become so naturally (i.e. they have been isolated as such from nature) or artificially (e.g. by breeding in vitro or by genetic manipulation). There will generally be at least one cell-type in which the replication-impaired viral vector can be grown - for example, modified vaccinia Ankara (MVA) can be grown in CEF cells. A vector may be selected from a human or simian adenovirus or a poxvirus vector. A viral vector may be a non-replicated adenoviral vector or a non-replicating modified vaccinia Ankara viral vector.

[0361] Typically, the viral vector is incapable of causing a significant infection in an animal subject, typically in a mammalian subject such as a human or other primate.

[0362] The vector(s) may be a DNA vector, such as a DNA plasmid. A DNA vector(s) may be capable of expression in a mammalian cell expression system, such as a CHO cell or an immunised cell. The vector may be suitable for expression in a bacterial and / or insect host cell or expression system, such as any of those exemplified herein. A non-limiting example of a suitable expression vector is a mammalian cell expression vector such as pcDNA3.1(+), pcDNA3.1(-), pcDNA3.1(+)-myc-His A, pcDNA3.1(+)-myc-His B, pcDNA3.1(+)-myc-His C, pcDNA3.1(-)-myc-His A, pcDNA3.1(-)-myc-His B, pcDNA3.1(-)-myc-His C, pCI-Neo, pcDNA3.1(+)-C-DYK, pcDNA3.1(+)-C-HA, pcDNA3.1(+)-C-6His, pcDNA3.1(+)-C-Myc, pcDNA3.1(+)-N-DYK, pcDNA3.1(+)-N-HA, pcDNA3.1(+)-N-6His, pcDNA3.1(+)-N-Myc, pcDNA3.1(+)-N-GST(Thrombin), pcDNA3.1(+)-N-GST(TEV), pcDNA3.1 / Hygro(+), pcDNA3.1 / Hygro(-), pcDNA3.1 / Zeo (+), pcDNA3.1 / Zeo (-), pCMV-3Tag-la, pCMV-3Tag-2a, pCMV-3Tag-3a, pCMV-3Tag-4a, pcDNA3.1(+)-C-eGFP, pcDNA3.1(+)-N-eGFP, pcDNA3.1(+)-P2A-eGFP, pcDNA3.1(+)-P2A, pcDNA3.1(+)-N-DYK-P2A, pcDNA3.1(+)-C-DYK-P2A, pCMV-3Tag-la-P2A, pCMV-3Tag-3a-P2A, pCDNA3.4 or pGenLenti. Another non-limiting example of a suitable expression vector is a yeast expression vector such as pAO815, pPIC 3.5k, pPIC9, pPIC9K, pPICZA, pPICZB, pPICZC, pPICZalphaA, pPICZalphaB, pPICZalphaC, pESC-TRP, pESC-URA, pESC-HIS or pESC-LEU. Yet another non-limiting example of a suitable expression vector is an insect cell expression vector such as pBacPAK8, pBacPAK9, pAcG2T, pAcHLT A, pAcHLT B, pAcHLT C, pAcGHLT A, pAcGHLT B, pAcGHLT C, pAcSG2, pBAC-1, pFastBac1, pFastBacHT-A, pFastBacHT-B, pFastBacHT-C or pFastBac-Dual. Yet another non-limiting example of a suitable expression vector is a bacterial expression vector such as pBluescript II KS(-), pBluescript II KS(+), pBluescript II SK(-), pBluescript II SK(+), pET-3a, pET-3b, pET-3c, pET-3d, pET-9a, pET-11a, pET-11b, pET-11c, pET-11d, pET-14b, pET-15b, pET-16b, pET-17b, pET-19b, pET-20b(+), pET-21a(+), pET-21b(+), pET-21d(+), pET-22b(+), pET-23a(+), pET-24a(+), pET-24a(+)-TEV, pET-24b(+), pET-24c(+), pET-24d(+), pET-25b(+), pET-26b(+), pET-27b(+), pET-28a(+), pET-28a(+)-TEV, pET-28b(+), pET-28c(+), pET-29a(+), pET-29b(+), pET-29c(+), pET-30a(+), pET-30b(+), pET-30c(+), pET-31b(+), pET-32a(+), pET-32b(+), pET-41a(+), pET-41b(+), pET-41c(+), pET-42a(+), pET-42b(+), pET-42c(+), pET-43.1a(+), pET-43.1b(+), pET-45b(+), pET-50b(+), pET-51b(+), pET-52b(+), pGEX-2TK, pGEX-4T-l, pGEX-4T-2, pGEX-4T-3, pGEX-5X-l, pGEX-5X-2, pGEX-5X-3, pGEX-6P-l, pGEX-6P-2, pGEX-6P-3, pMAL-c4x, pMAL-c5E, pMAL-c5x, pMAL-p5E, pMAL-p5G, pMAL-p5X, pQE-1, pQE-60, pGS-21a, pACYCDuet-1, pETDuet-1, pCDFDuet-1, pRSFDuet-1, pCOLADuet-1, pGEX-4T-l-H(RBS), pGEX-4T-l-M(RBS), pGEX-5X-l-H(RBS), pGEX-5X-l-M(RBS), pGEX-6P-l-H(RBS), pGEX-6P-l-M(RBS), pMAL-c4x-l-H(RBS), pMAL-c4x-l-M(RBS) or pGenDONR. The suitable expression vector may be optionally modified to encode an N-terminal and / or C-terminal tag, such as a hexa-histidine tag and / or a protease cleavage site, such as a TEV protease cleavage site. The vector(s) may be an RNA vector, such as a self-amplifying RNA vaccine (Geall, AJ. et al., Proc Natl Acad Sci USA 2012; 109(36) pp. 14604-9; incorporated herein by reference).

[0363] The present invention may be a phage vector, such as an AAV / phage hybrid vector as described in Hajitou et al., Cell 2006; 125(2) pp. 385-398; herein incorporated by reference.

[0364] A PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein or a VLP of the invention may include a leader sequence, for example to assist in recombinant production and / or secretion. Any suitable leader sequence may be used, including conventional leader sequences known in the art. Suitable leader sequences include Bip leader sequences, which are commonly used in the art to aid secretion from insect cells and human tissue plasminogen activator leader sequence (tPA), which is routinely also used in viral and DNA based vaccines and for protein vaccines to aid secretion from mammalian cell expression platforms. By way of a nonlimiting example, a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein or a VLP of the invention may include the secretory signal from bovine tissue plasminogen activator, or may include another signal to direct subcellular trafficking. Alternatively, a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein or a VLP of the invention may be the mature form in which the N-terminal signal peptide has been removed.

[0365] A PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein or a VLP of the invention may additionally comprise an N- and / or C-terminal tag, for example to assist in recombinant production and / or purification. Any N- and / or C-terminal tag may be used, including conventional tags known in the art. Suitable tags sequences include C-terminal hexa-histidine tags and the " C-tag" (the four amino acids EPEA at the C-terminus), which are commonly used in the art to aid purification from heterologous expression systems, e.g. insect cells, mammalian cells, bacteria, or yeast. Other examples of suitable tags include GST and MBP tags, or any other conventional tag which may be used to facilitate increased expression. In other embodiments, a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein or a VLP of the invention may be purified from heterologous expression systems without the need to use a purification tag.

[0366] A PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein or a VLP of the invention may be expressed using any suitable systems known to the person skilled in the art.

[0367] A PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein or a VLP of the invention may be expressed using conventional systems. Such a system may be a prokaryotic or a eukaryotic system. Examples of such systems are well-known in the art. Non-limiting examples of suitable host systems include Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris, non-lytic insect cell expression systems such as Schneider 2 (S2) and Schneider 3 (S3) cells from Drosophila melanogaster and Sf9 and Sf21 cells from Spodoptera frugiperda, and mammalian expression systems such as CHO cells and human embryonic kidney (HEK / HEK293) cells (preferably CHO cells).

[0368] Accordingly, the invention provides a host cell containing a recombinant expression vector which encodes an antibody of the invention. In a preferred embodiment the host cell is a CHO cell.

[0369] Viral vectors, RNA vectors and DNA plasmids of the invention may comprise any nucleic acid of the invention. The nucleic acid of the invention may be operably linked to a promoter. Any suitable promoter known in the art may be used, such as murine CMV promoter, EFla, PGK1, Ubc, human beta actin, CAG, TRE, Hl or U6, optionally murine CMV promoter (SEQ ID NO: 85).

[0370] A viral vector, RNA vector or DNA plasmid of the invention may further comprise a nucleic acid sequence encoding a VLP core (e.g. at least one VLP carrier subunit). Preferably, said nucleic acid encodes the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent.

[0371] Method of production

[0372] The invention further provides a method of producing a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein, or a VLP of the invention. Such method may comprise culturing a host cell (e.g. as described herein) under conditions suitable to express said PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, fusion protein, or VLP, and recovering the PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, fusion protein, or VLP produced.

[0373] The method of production may further comprise a step of introducing a nucleic acid molecule or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, fusion protein, or VLP (such in as those described herein) into the host cell. Alternatively or in addition, said method may further comprise purifying the PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, fusion protein, or VLP produced. Routine methods for the introduction of nucleic acid molecules or vectors (e.g. a viral vector, RNA vector or DNA plasmid) into cells are known in the art, as are suitable culture media and conditions, and method of purification. These can be selected by one of ordinary skill without undue burden.

[0374] Pharmaceutical and vaccine compositions

[0375] A PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, or a vector (e.g. a viral vector, RNA vector or DNA plasmid) can be combined or administered in addition to a pharmaceutically acceptable carrier.

[0376] Alternatively or in addition, a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, or a vector (e.g. a viral vector, RNA vector or DNA plasmid) can further be combined with one or more of a salt, excipient, diluent, adjuvant, immunoregulatory agent and / or antimicrobial compound.

[0377] The term "vaccine" is herein used interchangeably with the terms "therapeutic / prophylactic composition", "formulation" or "medicament".

[0378] The pharmaceutical and vaccine compositions of the invention can be combined or administered in addition to a pharmaceutically acceptable carrier. Alternatively or in addition the vaccine of the invention can further be combined with one or more of a salt, excipient, diluent, adjuvant, immunoregulatory agent and / or antimicrobial compound.

[0379] Pharmaceutically acceptable salts include acid addition salts formed with inorganic acids such as, for example, hydrochloric or phosphoric acids, or with organic acids such as acetic, oxalic, tartaric, maleic, and the like. Salts formed with the free carboxyl groups may also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, 2-ethylamino ethanol, histidine, procaine, and the like.

[0380] Administration of pharmaceutical and vaccine compositions, therapeutic formulations, medicaments and prophylactic formulations (e.g. vaccines) is generally by conventional routes e.g. intravenous, subcutaneous, intraperitoneal, or mucosal routes. The administration may be by parenteral injection, for example, a subcutaneous, intradermal or intramuscular injection. Formulations comprising neutralizing antibodies may be particularly suited to administration intravenously, intramuscularly, intradermally, or subcutaneously. Accordingly, immunogenic compositions, therapeutic formulations, medicaments and prophylactic formulations (e.g. vaccines) of the invention are typically prepared as injectables, either as liquid solutions or suspensions. Solid forms suitable for solution in, or suspension in, liquid prior to injection may alternatively be prepared. The preparation may also be emulsified, or the peptide encapsulated in liposomes or microcapsules.

[0381] The active immunogenic ingredients (such as a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, or a vector (e.g. a viral vector, RNA vector or DNA plasmid) of the invention) are often mixed with excipients which are pharmaceutically acceptable and compatible with the active ingredient. Suitable excipients are, for example, water, saline, dextrose, glycerol, ethanol, or the like and combinations thereof. In addition, if desired, the vaccine may contain minor amounts of auxiliary substances such as wetting or emulsifying agents, pH buffering agents, and / or adjuvants which enhance the effectiveness of the vaccine.

[0382] As exemplified herein, a composition of the invention may comprise a surfactant, such as Tween80 to reduce aggregation of the active ingredient (such as a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, or a vector (e.g. a viral vector, RNA vector or DNA plasmid) of the invention.

[0383] Generally, the carrier is a pharmaceutically-acceptable carrier. Non-limiting examples of pharmaceutically acceptable carriers include water, saline, and phosphate-buffered saline. In some embodiments, however, the composition is in lyophilized form, in which case it may include a stabilizer, such as BSA. In some embodiments, it may be desirable to formulate the composition with a preservative, such as thiomersal or sodium azide, to facilitate long term storage.

[0384] Examples of additional adjuvants which may be effective include but are not limited to: complete Freund's adjuvant (CFA), Incomplete Freund's adjuvant (IFA), Saponin, a purified extract fraction of Saponin such as Quil A, a derivative of Saponin such as QS-21, lipid particles based on Saponin such as ISCOM / ISCOMATRIX, E. coli heat labile toxin (LT) mutants such as LTK63 and / or LTK72, aluminium hydroxide, N-acetyl-muramyl-L-threonyl-D-isoglutamine (thr-MDP), N-acetyl-nor-muramyl-L-alanyl-D-isoglutamine (CGP 11637, referred to as nor-MDP), N-acetylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine-2-(l'-2'-dipalmitoyl-sn-glycero-3-hydroxyphosphoryl oxy)-ethylamine (CGP 19835A, referred to as MTP-PE), and RIBI, which contains three components extracted from bacteria, monophosphoryl lipid A, trehalose dimycolate and cell wall skeleton (MPL+TDM+CWS) in a 2% squalene / Tween 80 emulsion, the MF59 formulation developed by Novartis, the AS02, AS01, AS03 and AS04 adjuvant formulations developed by GSK Biologicals (Rixensart, Belgium) and AS05. Examples of buffering agents include, but are not limited to, sodium succinate (pH 6.5), and phosphate buffered saline (PBS; pH 6.5 and 7.5). The pH of a composition of the invention may be greater than pH 7. The pH of a composition of the invention may be pH 7.4 or greater, such as about pH 7.4, pH 7.5, pH 7.6, pH 7.7, pH 7.7, pH 7.8, pH 7.9, pH 8.0, pH 8.1, or pH 8.2. The pH of a composition of the invention may preferably be about pH 8. The pH of a composition of the invention may be between pH 7-8.5, such as between pH 7-8, between pH 7-8 or between pH 7.4-8. The pH of a composition of the invention may preferably be between pH 7.4-8.

[0385] Additional formulations which are suitable for other modes of administration include suppositories and, in some cases, oral formulations or formulations suitable for distribution as aerosols. For suppositories, traditional binders and carriers may include, for example, polyalkylene glycols or triglycerides; such suppositories may be formed from mixtures containing the active ingredient in the range of 0.5% to 10%, preferably l%-2%.

[0386] Oral formulations include such normally employed excipients as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, and the like. These compositions take the form of solutions, suspensions, tablets, pills, capsules, sustained release formulations or powders.

[0387] It is within the routine practice of a clinician to determine an effective amount of a pharmaceutical or vaccine composition of the invention. An effective amount is an amount sufficient to elicit a protective immune response against malaria. A clinician will also be able to determine appropriate dosage interval using routine skill.

[0388] In some preferred embodiments the adjuvant is selected from A01 (MF59 or biosimilar thereof), A02 (AS01 or biosimilar thereof, such as AS01B which is produced by GSK) and A05, each of which is exemplified herein. In some preferred embodiments the adjuvant is MF59 or a biosimilar thereof (comprising squalene, Tween 80 and Span). In some preferred embodiments the adjuvant is AS01 or a biosimilar thereof (comprising Q. S21, 3D-MPL, DOPC and cholesterol). In some preferred embodiments the adjuvant is A05 (comprising CpG, DOPC and cholesterol). In some preferred embodiments, the adjuvant is a combination of MF59 or a biosimilar thereof and AS01 or a biosimilar thereof.

[0389] The A01 (MF59) adjuvant formulation typically consists of: squalene (21 mg), Tween 80 (2.5 mg) and Span (852.5 mg) per 0.5 mL of sterile water making up the typical concentration of A01. The A02 (AS01B) adjuvant formulation typically consists of: QS21 (50 pg), 3D-MPL (50 pg), DOPC (1 mg) and cholesterol (0.25 mg) per 0.5 mL of sterile water making up the typical concentration of A02. The A05 adjuvant formulation typically consists of: CpG (50 pg), DOPC (2.18 mg) and cholesterol (0.33 mg) per 0.5 mLof sterile water making up the typical concentration of A05. The amount of adjuvant (and hence of the components therein) administered may be readily determined by one of ordinary skill in the art without undue burden. By way of non-limiting example, in the Examples herein, 50 pl of the above A02 and A05 adjuvant formulations are administered with 1 pl of antigen (VLP or fusion protein) as a full dose of the antigen, and for a half dose, the initial formulation was diluted x2 and 50 pl of the diluted formulation administered. By way of a further non-limiting example, in the Examples herein, a stock solution of 2x the above A01 adjuvant formulation was prepared and 25 pl administered with 1 pl of antigen (VLP or fusion protein) as a full dose of the antigen.

[0390] The adjuvant may be used at the typical concentration above or at a lower concentration, optionally the adjuvant may be at a concentration of about 75%, about 50% or about 25% of the typical concentration, preferably the adjuvant may be at a concentration of about 50% of the typical concentration. In some preferred embodiments the adjuvant is A01 or a biosimilar thereof at a concentration of about 50% of the typical concentration. In some preferred embodiments the adjuvant is A02 or a biosimilar thereof at a concentration of about 50% of the typical concentration. In some preferred embodiments the adjuvant is A05 or a biosimilar thereof at a concentration of about 50% of the typical concentration.

[0391] A pharmaceutical or vaccine composition of the invention may be stable at 4°C for at least one week, at least two weeks, at least four weeks, at least 8 weeks, at least 12 weeks or more. Alternatively or in addition, a pharmaceutical or vaccine composition of the invention may be stable at 25°C for at least one week, at least two weeks, at least four weeks, at least 8 weeks, at least 12 weeks or more. Further alternatively or in addition, a pharmaceutical or vaccine composition of the invention may be stable at 37°C for at least one week, at least two weeks, at least four weeks, at least 8 weeks, at least 12 weeks or more.

[0392] A pharmaceutical or vaccine composition of the invention may be stable at 4°Cfor at least 2 weeks. A pharmaceutical or vaccine composition of the invention may be stable at 25°C for at least 2 weeks. A pharmaceutical or vaccine composition of the invention may be stable at 37°C for at least 2 weeks. A pharmaceutical or vaccine composition of the invention may be stable at 4°C, 25°C and 37°C for at least 2 weeks.

[0393] A pharmaceutical or vaccine composition of the invention may be stable at -80°C for at least one month, at least two months, at least three months, at least four months, at least 8 months, at least 12 months or more. Typically, a pharmaceutical or vaccine composition of the invention is stable at -80°C for at least 3 months.

[0394] A pharmaceutical or vaccine composition of the invention may be a liquid formulation. A pharmaceutical or vaccine composition of the invention may be a solid formulation, such as a lyophilised formulation.

[0395] Co-administration and pharmaceutical preparations

[0396] The above-described PvCSP proteins or immunogenic fragments (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragments thereof, PvDBPII or immunogenic fragment thereof, fusion proteins, VLPs, isolated nucleic acid molecules, vectors (e.g. a viral vector, RNA vector or DNA plasmid) and vaccine compositions may be co-administered. The PvCSP proteins or immunogenic fragments (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragments thereof, PvDBPII or immunogenic fragment thereof, fusion proteins, VLPs, isolated nucleic acid molecules, vectors (e.g. a viral vector, RNA vector or DNA plasmid) and vaccine compositions to be co-administered may be in the same form or different forms. By way of non-limiting example, a vaccine composition comprising a VLP comprising a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) may be co-administered with a vaccine composition comprising a VLP comprising Pvs230 or immunogenic fragment thereof. By way of further non-limiting example, a vaccine composition comprising a VLP comprising a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) may be co-administered with a vaccine composition comprising a VLP comprising PvDBPII or immunogenic fragment thereof. By way of further non-limiting example, a vaccine composition comprising a VLP comprising a Pvs230 protein or immunogenic fragment thereof may be co-administered with a vaccine composition comprising a VLP comprising PvDBPII or immunogenic fragment thereof. By way of further non-limiting example, a vaccine composition comprising a VLP comprising a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) and a Pvs230 or immunogenic fragment thereof may be co-administered with a vaccine composition comprising a VLP comprising PvDBPII or immunogenic fragment thereof. By way of further non-limiting example, a vaccine composition comprising a VLP comprising a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) and a PvDBPII or immunogenic fragment thereof may be co-administered with a vaccine composition comprising a VLP comprising Pvs230 or immunogenic fragment thereof. By way of further non-limiting example, a vaccine composition comprising a VLP comprising a Pvs230 protein or immunogenic fragment thereof and a PvDBPII or immunogenic fragment thereof may be co-administered with a vaccine composition comprising a VLP comprising a PvCSP or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof). By way of a further non-limiting example, a vaccine composition comprising a DNA plasmid encoding a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) may be co-administered with a vaccine composition comprising a VLP comprising Pvs230 or immunogenic fragment thereof. By way of further non-limiting example, a vaccine composition comprising a DNA plasmid encoding a VLP comprising a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) may be co-administered with a vaccine composition comprising a DNA plasmid encoding a VLP comprising PvDBPII or immunogenic fragment thereof. By way of further non-limiting example, a vaccine composition comprising a DNA plasmid encoding a VLP comprising a Pvs230 protein or immunogenic fragment thereof may be co-administered with a vaccine composition comprising a DNA plasmid encoding a VLP comprising PvDBPII or immunogenic fragment thereof. By way of further non-limiting example, a vaccine composition comprising a DNA plasmid encoding a VLP comprising a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) and a Pvs230 or immunogenic fragment thereof may be co-administered with a vaccine composition comprising a DNA plasmid encoding a VLP comprising PvDBPII or immunogenic fragment thereof. By way of further non-limiting example, a vaccine composition comprising a DNA plasmid encoding a VLP comprising a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) and a PvDBPII or immunogenic fragment thereof may be co-administered with a vaccine composition comprising a DNA plasmid encoding a VLP comprising Pvs230 or immunogenic fragment thereof. By way of further non-limiting example, a vaccine composition comprising a DNA plasmid encoding a VLP comprising a Pvs230 protein or immunogenic fragment thereof and a PvDBPII or immunogenic fragment thereof may be co-administered with a vaccine composition comprising a DNA plasmid encoding a VLP comprising a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof).

[0397] In particular, a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a VLP comprising said PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), or an isolated nucleic acid molecule, or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) may be co-administered with a Pvs230 or immunogenic fragment thereof, a VLP comprising said Pvs230 protein or immunogenic fragment thereof, or an isolated nucleic acid molecule, or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said Pvs230 protein or immunogenic fragment thereof.

[0398] In particular, a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a VLP comprising said PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), or an isolated nucleic acid molecule, or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) may be co-administered with a PvDBPII or immunogenic fragment thereof, a VLP comprising said PvDBPII protein or immunogenic fragment thereof, or an isolated nucleic acid molecule, or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said PvDBPII protein or immunogenic fragment thereof.

[0399] In particular, a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a VLP comprising said PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), or an isolated nucleic acid molecule, or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) may be co-administered with (i) a PvDBPII or immunogenic fragment thereof, a VLP comprising said PvDBPII protein or immunogenic fragment thereof, or an isolated nucleic acid molecule, or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said PvDBPII protein or immunogenic fragment thereof; and (ii) a Pvs230 or immunogenic fragment thereof, a VLP comprising said Pvs230 protein or immunogenic fragment thereof, or an isolated nucleic acid molecule, or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said Pvs230 protein or immunogenic fragment thereof.

[0400] The PvCSP proteins or immunogenic fragments (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragments thereof, PvDBPII or immunogenic fragment thereof, fusion proteins, VLPs, isolated nucleic acid molecules, vectors (e.g. a viral vector, RNA vector or DNA plasmid) and vaccine compositions for co-administration may be formulated in the same or separate vaccine compositions. When formulated as separate vaccine compositions, the vaccine compositions may be administered simultaneously, separately or sequentially. By way of non-limiting example, a vaccine composition comprising a VLP comprising a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) may be co-administered with a vaccine composition comprising a VLP comprising Pvs230 or immunogenic fragment thereof, wherein said compositions are administered sequentially. By way of a further non-limiting example, a vaccine composition comprising a DNA plasmid encoding a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof) may be co-administered with a vaccine composition comprising a VLP comprising Pvs230 or immunogenic fragment thereof, wherein said compositions are administered simultaneously.

[0401] Wherein PvCSP proteins or immunogenic fragments (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragments thereof, PvDBPII or immunogenic fragment thereof, fusion proteins, VLPs, isolated nucleic acid molecules, vectors (e.g. a viral vector, RNA vector or DNA plasmid) and vaccine compositions of the invention are co-administered simultaneously, they may be administered in a single composition or as separate compositions.

[0402] Wherein the PvCSP proteins or immunogenic fragments (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragments thereof, PvDBPII or immunogenic fragment thereof, fusion proteins, VLPs, isolated nucleic acid molecules, vectors (e.g. a viral vector, RNA vector or DNA plasmid) or vaccine compositions for co-administration in a form suitable for simultaneous, separate or sequential use, these may be referred to as pharmaceutical preparations.

[0403] Thus, pharmaceutical preparations are combined preparations which may comprise or consist of two or more of the PvCSP proteins or immunogenic fragments (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragments thereof, PvDBPII or immunogenic fragment thereof, fusion proteins, VLPs, isolated nucleic acid molecules, and vectors (e.g. a viral vector, RNA vector or DNA plasmid) of the invention in a form suitable for simultaneous, separate or sequential use. Typically, a pharmaceutical preparation comprises (i) a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a VLP comprising said PvCSP protein or immunogenic fragment thereof or an isolated nucleic acid or vector encoding said PvCSP protein or immunogenic fragment thereof; and (ii) a Pvs230 protein or immunogenic fragment thereof, a VLP comprising said Pvs230 protein or immunogenic fragment thereof or an isolated nucleic acid or vector encoding said Pvs230 protein or immunogenic fragment thereof; in a form suitable for simultaneous, separate or sequential use. Alternatively, a pharmaceutical preparation comprises (i) a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a VLP comprising said PvCSP protein or immunogenic fragment thereof or an isolated nucleic acid or vector encoding said PvCSP protein or immunogenic fragment thereof; and (ii) a PvDBPII protein or immunogenic fragment thereof, a VLP comprising said PvDBPII protein or immunogenic fragment thereof or an isolated nucleic acid or vector encoding said PvDBPII protein or immunogenic fragment thereof; in a form suitable for simultaneous, separate or sequential use. Alternatively, a pharmaceutical preparation comprises (i) a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a VLP comprising said PvCSP protein or immunogenic fragment thereof or an isolated nucleic acid or vector encoding said PvCSP protein or immunogenic fragment thereof; and (ii) a Pvs230 protein or immunogenic fragment thereof, a VLP comprising said Pvs230 protein or immunogenic fragment thereof or an isolated nucleic acid or vector encoding said Pvs230 protein or immunogenic fragment thereof; and (iii) a PvDBPII protein or immunogenic fragment thereof, a VLP comprising said PvDBPII protein or immunogenic fragment thereof or an isolated nucleic acid or vector encoding said PvDBPII protein or immunogenic fragment thereof; in a form suitable for simultaneous, separate or sequential use.

[0404] A pharmaceutical preparation may comprise or consist of:

[0405] (a) a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a VLP comprising said PvCSP protein or immunogenic fragment thereof or an isolated nucleic acid or vector encoding said PvCSP protein or immunogenic fragment thereof; and / or

[0406] (b) a Pvs230 protein or immunogenic fragment thereof, a VLP comprising said Pvs230 protein or immunogenic fragment thereof or an isolated nucleic acid or vector encoding said Pvs230 protein or immunogenic fragment thereof;

[0407] and one or more additional P. vivax polypeptide, or a variant, derivative or fragment thereof, or a nucleic acid encoding said polypeptide; in a form suitable for simultaneous, separate or sequential use. Nonlimiting examples of additional P. vivax polypeptides include PvDBPII and PvRBP2b.

[0408] In particular, a pharmaceutical preparation may comprise or consist of:

[0409] (a) a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a VLP comprising said PvCSP protein or immunogenic fragment thereof or an isolated nucleic acid or vector encoding said PvCSP protein or immunogenic fragment thereof; and / or (b) a Pvs230 protein or immunogenic fragment thereof, a VLP comprising said Pvs230 protein or immunogenie fragment thereof or an isolated nucleic acid or vector encoding said Pvs230 protein or immunogenic fragment thereof; and / or

[0410] (c) a PvDBPII protein or immunogenic fragment thereof, a VLP comprising said PvDBPII protein or immunogenic fragment thereof or an isolated nucleic acid or vector encoding said PvDBPII protein or immunogenic fragment thereof.

[0411] A pharmaceutical preparation of the invention may further comprise one or more additional P. vivax polypeptide, ora variant, derivative or fragment thereof, or a nucleic acid encoding said polypeptide.

[0412] Alternatively or in addition, a pharmaceutical preparation of the invention may further comprise one or more additional P. falciparum polypeptide, or a variant, derivative or fragment thereof, or a nucleic acid encoding said polypeptide. Non-limiting examples of P. falciparum polypeptides include PfCyRPA, PfRH5, PfRIPR, PfP113, PfRhopH3, PfRAP2, PfAMAl, PfRON2, PfEBA175, PfRHl, PfRH2a, PfRH2b, PfRH4, PfRAPl, PfRAP3, PfMSRPS, PfRAMA, PfSERA9, PfEBA181, PfCSS, PfPTRAMP, PfAARP PfCSP and Pf230. The one or more additional P. falciparum polypeptide, or a variant, derivative or fragment thereof, or a nucleic acid encoding said polypeptide may be a fusion protein comprising a combination of two or more of PfCyRPA, PfRH5, PfRIPR, PfP113, PfRhopH3, PfRAP2, PfAMAl, PfRON2, PfEBA175, PfRHl, PfRH2a, PfRH2b, PfRH4, PfRAPl, PfRAP3, PfMSRPS, PfRAMA, PfSERA9, PfEBA181, PfCSS, PfPTRAMP, PfAARP PfCSP and Pf230. The one or more additional P. falciparum polypeptide, or a variant, derivative or fragment thereof, or fusion protein or a nucleic acid encoding said polypeptide or fusion protein may be displayed on a VLP. In some preferred embodiments, the one or more additional P. falciparum polypeptide, or a variant, derivative or fragment thereof may be PfCSP and / or Pf230, preferably both PfCSP and Pf230. These may be presented in a fusion protein or VLP form, analogous to the fusion proteins and VLPs comprising PvCSP and / or Pv230, as described herein.

[0413] According to the invention pharmaceutical preparations are in a form suitable for simultaneous, separate or sequential use of the pharmaceutical or vaccine compositions of the invention.

[0414] Therapeutic Indications

[0415] The invention provides a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, a vector (e.g. a viral vector, RNA vector or DNA plasmid), or a composition for use in therapy.

[0416] In particular, the invention provides a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, a vector (e.g. a viral vector, RNA vector or DNA plasmid), or a composition for use in a method of preventing and / or treating malaria. The present invention also provides a method of stimulating or inducing an immune response in a subject comprising administering to the subject a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, a vector (e.g. a viral vector, RNA vector or DNA plasmid), or a composition of the invention (as described above).

[0417] Thus, the invention provides a composition comprising a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, a vector (e.g. a viral vector, RNA vector or DNA plasmid), or a composition of the invention.

[0418] The method of stimulating or inducing an immune response in a subject may comprise administering a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, a vector (e.g. a viral vector, RNA vector or DNA plasmid), or a composition of the invention (as described above) to a subject.

[0419] In the context of the therapeutic uses and methods, a "subject" is any animal subject that would benefit from stimulation or induction of an immune response against a Plasmodium parasite, particularly a Plasmodium vivax parasite. Typical animal subjects are mammals, such as primates, for example, humans.

[0420] Thus, the present invention provides a method for treating or preventing malaria.

[0421] The present invention also provides a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, a vector (e.g. a viral vector, RNA vector or DNA plasmid), or a composition of the invention (as described above) for use in the prevention or treatment of malaria.

[0422] The present invention also provides the use of a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, a vector (e.g. a viral vector, RNA vector or DNA plasmid), or a composition of the invention (as described above) in the manufacture of a medicament for the prevention and / or treatment of malaria.

[0423] The present invention provides the use of a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, a vector (e.g. a viral vector, RNA vector or DNA plasmid), or a composition of the invention (as described above), for use either alone or in combination in the prevention or treatment of malaria.

[0424] The method for treating or preventing malaria may comprise administering a therapeutically effective amount of a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, a fusion protein, PvDBPII protein or immunogenic fragment thereof, a VLP, an isolated nucleic acid molecule, a vector (e.g. a viral vector, RNA vector or DNA plasmid), or a composition of the invention (as described above), either alone or in combination, to a subject.

[0425] As used herein, the term "treatment" or "treating" embraces therapeutic or preventative / prophylactic measures and includes post-infection therapy and amelioration of malaria.

[0426] As used herein, the term "preventing" includes preventing the initiation of malaria and / or reducing the severity or intensity of malaria. The term "preventing" includes inducing or providing protective immunity against malaria. Immunity to malaria may be quantified using any appropriate technique, examples of which are known in the art.

[0427] A PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, a vector (e.g. a viral vector, RNA vector or DNA plasmid), or a composition of the invention (as described above) may be administered to a subject (typically a mammalian subject such as a human or other primate) already having malaria, a condition or symptoms associated with malaria, to treat or prevent malaria. For example, the subject may be suspected of having come in contact with Plasmodium parasite (particularly P. vivax) or has had known contact with Plasmodium parasite (particularly P. vivax) but is not yet showing symptoms of exposure.

[0428] When administered to a subject (e.g. a mammal such as a human or other primate) that already has malaria, or is showing symptoms associated with Plasmodium parasite (particularly P. vivax) infection, a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, a vector (e.g. a viral vector, RNA vector or DNA plasmid), or a composition of the invention (as described above) can cure, delay, reduce the severity of, or ameliorate one or more symptoms, and / or prolong the survival of a subject beyond that expected in the absence of such treatment.

[0429] Alternatively, a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, a vector (e.g. a viral vector, RNA vector or DNA plasmid), or a composition of the invention (as described above) may be administered to a subject (e.g. a mammal such as a human or other primate) who ultimately may be infected with Plasmodium parasite (particularly P. vivax), in order to prevent, cure, delay, reduce the severity of, or ameliorate one or more symptoms of malaria, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment, or to help prevent that subject from transmitting malaria.

[0430] The treatments and preventative therapies of the present invention are applicable to a variety of different subjects of different ages. In the context of humans, the therapies are applicable to children (e.g. infants, children under 5 years old, older children or teenagers) and adults. In the context of other animal subjects (e.g. mammals such as primates), the therapies are applicable to immature subjects and mature / adult subjects.

[0431] The present invention provides a composition comprising a PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, a fusion protein, a VLP, an isolated nucleic acid molecule, or a vector (e.g. a viral vector, RNA vector or DNA plasmid) of the invention (as described above). Said compositions may further comprise one or more further components as described herein. Said one or more further components may be one or more further PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), Pvs230 or immunogenic fragment thereof, PvDBPII protein or immunogenic fragment thereof, fusion protein, VLP, isolated nucleic acid molecule, or vector (e.g. a viral vector, RNA vector or DNA plasmid) as described above. Alternatively, or in addition, said one or more further component may be one or more pharmaceutically acceptable carrier, excipient or other component as described herein.

[0432] A PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a fusion protein or VLP comprising said PvCSP protein or immunogenic fragment thereof, or an isolated nucleic acid molecule or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said PvCSP protein or immunogenic fragment thereof is typically immunogenic in vivo and can induce high titres of antibodies against said PvCSP protein or immunogenic fragment thereof. A PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a fusion protein or VLP comprising said PvCSP protein or immunogenic fragment thereof, or an isolated nucleic acid molecule or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said PvCSP protein or immunogenic fragment thereof may induce antibodies with a geometric mean titre (GMT) of at least 15,000 lU / mL, optionally after boosting with a second dose of the PvCSP protein or immunogenic fragment thereof (e.g. any chimeric PvCSP protein or immunogenic fragment thereof), a fusion protein or VLP comprising said PvCSP protein or immunogenic fragment thereof, or an isolated nucleic acid molecule or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said PvCSP protein or immunogenic fragment thereof. Said GMT may be measured at any appropriate time point, such as 42 days post-administration of a first dose, as exemplified herein. A VLP comprising said PvCSP protein or immunogenic fragment thereof, or an isolated nucleic acid molecule or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said VLP, may induce antibodies with a geometric mean titre (GMT) of at least at least 50,000 lU / mL against PvCSP or the immunogenic fragment thereof, optionally at least 60,000 lU / mL, optionally after boosting with a second dose of the VLP comprising said PvCSP protein or immunogenic fragment thereof, or an isolated nucleic acid molecule or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said VLP. Said GMT may be measured at any appropriate time point, such as 42 days post-administration of a first dose, as exemplified herein. Said GMT may also be measured at any appropriate time point, such as 2-3 weeks post-administration of a first, second or third dose, as exemplified herein.

[0433] A Pvs230 protein or immunogenic fragment, a fusion protein or VLP comprising said Pvs230 protein or immunogenic fragment thereof, or an isolated nucleic acid molecule or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said Pvs230 protein or immunogenic fragment thereof is typically immunogenic in vivo and can induce high titres of antibodies against said Pvs230 protein or immunogenic fragment thereof. A Pvs230 protein or immunogenic fragment thereof, a fusion protein or VLP comprising said Pvs230 protein or immunogenic fragment thereof, or an isolated nucleic acid molecule or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said Pvs230 protein or immunogenic fragment thereof may induce antibodies with a geometric mean titre (GMT) of at least 2,000 lU / mL, optionally after boosting with a second dose of the Pvs230 protein or immunogenic fragment thereof, a fusion protein or VLP comprising said Pvs230 protein or immunogenic fragment thereof, or an isolated nucleic acid molecule or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said Pvs230 protein or immunogenic fragment thereof. Said GMT may be measured at any appropriate time point, such as 42 days post-administration of a first dose, as exemplified herein. A VLP comprising said Pvs230 protein or immunogenic fragment thereof, or an isolated nucleic acid molecule or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said VLP, may induce antibodies with a geometric mean titre (GMT) of at least at least 5,000 lU / mL against Pvs230 or the immunogenic fragment thereof, optionally at least 6,000 lU / mL, at least 7,000 lU / mL, or at least 8,000 lU / mL; optionally after boosting with a second dose of the VLP comprising said Pvs230 protein or immunogenic fragment thereof, or an isolated nucleic acid molecule or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said Pvs230. Said GMT may be measured at any appropriate time point, such as 42 days post-administration of a first dose, as exemplified herein. Said GMT may also be measured at any appropriate time point, such as 2-3 weeks post-administration of a first, second or third dose, as exemplified herein.

[0434] A PvDBPII protein or immunogenic fragment, a fusion protein or VLP comprising said PvDBPII protein or immunogenic fragment thereof, or an isolated nucleic acid molecule or vector (e.g. a viral vector, RNA vector or DNA plasmid) encoding said PvDBPII protein or immunogenic fragment thereof is typically immunogenic in vivo and can induce high titres of antibodies against said ...

Claims

CLAIMS1. A fusion protein comprising:(a) a Plasmodium vivax circumsporozoite protein (PvCSP) or immunogenic fragment thereof; and(b) a P. vivax 230 kDa sexual-stage protein (Pvs230) or immunogenic fragment thereof.

2. A fusion protein according to claim 1, wherein the PvCSP protein or immunogenic fragment thereof comprises:(a) at least 10 repeat motifs derived from the central repeat region of a Type I (VK210) of P. vivax;and / or(b) at least 10 repeat motifs derived from the central repeat region of a Type II (VK247) of P. vivax.

3. A fusion protein according to claim 1 or 2, wherein:(a) the at least 10 repeat motifs derived from the central repeat region of a Type I PvCSP comprise at least 5 copies of the amino acid sequence GDRADGQPA (SEQ ID NO: 4) and at least 5 copies of the amino acid sequence GDRAAGQPA (SEQ ID NO: 5), wherein optionally: (i) at least one copy of SEQ ID NO: 5 is interspersed between copies of SEQ ID NO: 4; and / or (ii) the at least 10 repeat motifs derived from the central repeat region of a Type I PvCSP further comprise at least one copy of GNGAGGQAA (SEQ ID NO: 6); and / or(b) the at least 10 repeat motifs derived from the central repeat region of a Type II PvCSP comprise at least 5 copies of the amino acid sequence ANGAGNQPG (SEQ ID NO: 7) and at least one copy of any two or more of (i) EDGAGNQPG (SEQ ID NO: 8); (ii) ANGAGDQPG (SEQ ID NO: 9); (iii) ANGADDQPG (SEQ ID NO: 10); (iv) ANGAGGQAA (SEQ ID NO: 11); and (v) GGNA ANKKAEDA GGNA GGNA (SEQ ID NO: 12).

4. A fusion protein according to any one of the preceding claims, wherein the PvCSP protein or immunogenic fragment thereof comprises:(a) an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 1; and / or(b) an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 2.

5. A fusion protein according to any one of the preceding claims, wherein the PvCSP protein or immunogenic fragment thereof comprises:(a) the amino acid sequence of SEQ ID NO: 1; and / or(b) the amino acid sequence SEQ ID NO: 2.

6. A fusion protein according to any one of the preceding claims, wherein the PvCSP protein or immunogenic fragment thereof is a chimeric PvCSP protein or immunogenic fragment thereof which comprises (a) at least 10 repeat motifs derived from the central repeat region of a Type I PvCSP; and (b) at least 10 repeat motifs derived from the central repeat region of a Type II PvCSP; wherein optionally said PvCSP protein or immunogenic fragment is as defined in any one of claims 3 to 5.

7. A fusion protein according to any one of the preceding claims, wherein the chimeric PvCSP protein or immunogenic fragment thereof comprises an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 3 or a fragment thereof which is at least 50% of the length of SEQ ID NO: 3.

8. A fusion protein according to any one of the preceding claims, wherein the PvCSP protein or immunogenic fragment thereof comprises an amino acid sequence of SEQ ID NO: 3.

9. A fusion protein according to any one of the preceding claims, wherein the PvCSP protein further comprises a PvCSP protein C-terminal region (CTR) or immunogenic fragment thereof.

10. A fusion protein according to claim 9, wherein the PvCSP protein CTR or immunogenic fragment thereof comprises an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 90.

11. A fusion protein according to claim 9 or 10, wherein the PvCSP protein CTR or immunogenic fragment thereof comprises an amino acid sequence of SEQ ID NO: 90.

12. A fusion protein according to claims 9 to 11, wherein the PvCSP protein or immunogenic fragment thereof comprises an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 91.

13. A fusion protein according to claims 9 to 12, wherein the PvCSP protein or immunogenic fragment thereof comprises the amino acid sequence of SEQ ID NO: 91.

14. A fusion protein according to any one of the preceding claims, wherein the Pvs230 or immunogenic fragment thereof comprises Domain 1 (DI) of PVs230, optionally with one or two substitutions.

15. A fusion protein according to any one of the preceding claims, wherein the Pvs230 or immunogenic fragment thereof comprises an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 13 or 83, preferably SEQ. ID NO: 13; or a fragment thereof which is at least 50% of the length of SEQ. ID NO: 13 or 83, preferably SEQ ID NO: 13.

16. A fusion protein according to any one of the preceding claims, wherein the Pvs230 or immunogenic fragment thereof comprises the amino acid sequence of SEQ ID NO: 13 or 83, preferably SEQ ID NO: 13.

17. A fusion protein according to any one of the preceding claims, wherein:(a) the PvCSP protein or immunogenic fragment thereof is N-terminal to Pvs230 or the immunogenic fragment thereof; or(b) the PvCSP protein or immunogenic fragment thereof is C-terminal to Pvs230 or the immunogenic fragment thereof;preferably wherein the PvCSP protein or immunogenic fragment thereof is N-terminal to the Pvs230 or the immunogenic fragment thereof.

18. A fusion protein according to any one of the preceding claims, further comprising a Plasmodium vivax Duffy binding protein II (PVDBPI I) or immunogenic fragment thereof.

19. A fusion protein according to claim 18, wherein the DBPII or immunogenic fragment thereof comprises an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 92.

20. A fusion protein according to claim 18 or 19, wherein the PVDBPII or immunogenic fragment thereof comprises the amino acid sequence of SEQ ID NO: 92, optionally comprising one, two or three substitutions; further optionally wherein the DBPII or immunogenic fragment thereof comprises the amino acid sequence of SEQ ID NO: 93.

21. A fusion protein according to any one of claims 18 to 20, wherein:(a) (i) the PVDBPII protein or immunogenic fragment thereof is N-terminal to Pvs230 or the immunogenic fragment thereof; or(ii) the PVDBPII protein or immunogenic fragment thereof is C-terminal to Pvs230 or the immunogenic fragment thereof;and / or(b) (i) the PVDBPII protein or immunogenic fragment thereof is N-terminal to PvCSP or the immunogenic fragment thereof; or(ii) the PVDBPII protein or immunogenic fragment thereof is C-terminal to PvCSP or the immunogenic fragment thereof;preferably wherein the PVDBPII protein or immunogenic fragment thereof is N-terminal to Pvs230 or the immunogenic fragment thereof and C-terminal to PvCSP or the immunogenic fragment thereof.

22. A fusion protein according to any one of the preceding claims, wherein the PvCSP protein or immunogenic fragment thereof, Pvs230 or the immunogenic fragment thereof and / or PVDBPII protein or immunogenic fragment thereof is separated from the remainder of the fusion protein by a linker polypeptide, wherein optionally the linker polypeptide comprises the amino acid sequence of SEQ ID NO: 100.

23. A fusion protein according to any one of the preceding claims, which further comprises a signal peptide sequence located at the N-terminus, optionally wherein the signal peptide sequence is selected from the group consisting of: SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16.

24. A fusion protein according to any one of the preceding claims, which further comprises a first complexing motif, optionally at the N- or C-terminus, preferably the C-terminus, and wherein optionally the first complexing motif comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 17, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36 and SEQ ID NO: 37.

25. A fusion protein according to claim 14, wherein the first complexing motif is separated from the remainder of the fusion protein by a linker polypeptide, wherein optionally the linker polypeptide comprises the amino acid sequence of SEQ ID NO: 18, SEQ ID NO: 101, SEQ ID NO: 115 and / or SEQ ID NO: 19.

26. A fusion protein according to any one of the preceding claims, which further comprises a label at the N-or C-terminus, wherein optionally said label is selected from the group consisting of a histag (SEQ ID NO: 38), GST (SEQ ID NO: 39), MBP (SEQ ID NO: 40), NusA (SEQ ID NO: 41) and SUMO (SEQ ID NO: 42), preferably SEQ ID NO: 38.

27. A fusion protein according to any one of the preceding claims, which comprises an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to an amino acid sequence selected from the group consisting of: SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126; preferably SEQ ID NO: 20, 21, 22, 23, 24, 94, 99 or 102-107; more preferably SEQ ID NO: 20, 24, 94, 99 or 102-107.

28. A fusion protein according to any one of the preceding claims, which comprises an amino acid sequence selected from the group consisting of: SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126; preferably SEQ ID NO: 20, 21, 22, 23,24, 94, 99 or 102-107; more preferably SEQ ID NO: 20, 24, 94, 99 or 102-107.

29. A chimeric PvCSP protein or immunogenic fragment thereof, which comprises (a) at least 10 repeat motifs derived from the central repeat region of a Type I PvCSP; and (b) at least 10 repeat motifs derived from the central repeat region of a Type II PvCSP; wherein said PvCSP protein or immunogenic fragment is as defined in any one of claims 3 to 5, and preferably said PvCSP protein or immunogenic fragment comprises an amino acid sequence of SEQ ID NO: 3, 56 to 62 or 84.

30. A chimeric PvCSP protein or immunogenic fragment thereof, which comprises (a) at least 10 repeat motifs derived from the central repeat region of a Type I PvCSP; (b) at least 10 repeat motifs derived from the central repeat region of a Type II PvCSP; and (c) a PvCSP protein C-terminal region (CTR) or immunogenic fragment thereof; wherein said PvCSP protein or immunogenic fragment is as defined in any one of claims 9 to 13, and preferably said PvCSP protein or immunogenic fragment comprises an amino acid sequence of SEQ ID NO: 91 or 111-114.

31. A virus-like particle (VLP) comprising:(a) at least one fusion protein as defined in any one of claims 1 to 28;(b) at least one chimeric PvCSP protein or immunogenic fragment thereof as defined in claim 29 or 30; and / or(c) at least one Pvs230 or immunogenic fragment thereof as defined in any one of claims 14 to 16; and / or(d) at least one PVDBPII protein or immunogenic fragment thereof as defined in any one of claims 19 or 20.

32. A VLP according to claim 31, wherein:(a) the VLP comprises a core which comprises at least one subunit of a VLP carrier, optionally the at least one subunit of the VLP carrier is selected from the group consisting of: an amino acid sequence of SEQ ID NO: 28, Hepatitis B surface antigen (SEQ ID NO: 43), HPV 18 LI protein (SEQ ID NO: 44), Hepatitis E P239 protein (SEQ ID NO: 45), HPV 16 LI protein (SEQ ID NO: 46), AP205 (SEQ ID NO: 47), ferritin (SEQ ID NO: 48), I53-50A (SEQ ID NO: 49) or I53-50B (SEQ ID NO: 69), preferably an amino acid sequence of SEQ ID NO: 28; and / or(b) the at least one fusion protein, at least one chimeric PvCSP protein or immunogenic fragment thereof and / or at least one Pvs230 or immunogenic fragment thereof is covalently linked to the at least one subunit of the VLP carrier, such that the at least one fusion protein, at least one chimeric PvCSP protein or immunogenic fragment thereof and / or at least one Pvs230 or immunogenic fragment thereof is presented on the surface of the VLP.

33. A VLP according to claim 31 or 32, wherein the at least one subunit of the VLP carrier further comprises a second complexing motif at the N- or C-terminus of the at least one subunit of the VLP carrier, preferably at the N-terminus of the at least one subunit of the VLP carrier; wherein optionally the second complexing motif comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 29, pilin-N (SEQ ID NO: 50), pilin-C (SEQ ID NO: 51), SnoopCatcher (SEQ ID NO: 52), SdyCatcher (SEQ ID NO: 53), SilkCatcher (SEQ ID NO: 54) or DogCatcher (SEQ ID NO: 55), preferably SEQ ID NO: 29.

34. A VLP according to claim 33, wherein the second complexing motif is separated from the remainder of the at least one subunit of the VLP carrier by a linker polypeptide, wherein optionally the linker polypeptide comprises the amino acid sequence of SEQ ID NO: 18 and / or SEQ ID NO: 19.

35. A VLP according to any one of claims 31 to 34, wherein the VLP core comprises an amino acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent.

36. A VLP according to any one of claims 31 to 35, wherein the VLP core comprises the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent.

37. A VLP according to any one of claims 33 to 36, wherein the first complexing motif and the second complexing motif are covalently linked, such that the at least one fusion protein, at least one chimeric PvCSP protein or immunogenic fragment thereof; at least one Pvs230 or immunogenic fragment thereof; and / or at least one PVDBPII protein or immunogenic fragment thereof is presented on the surface of the VLP.

38. A VLP according to any one of claims 33 to 37, wherein:(a) the fusion protein comprises an amino acid sequence selected from the group consisting of:SEQ ID NOs: 20 to 27, 63 to 66, 94-99, 102-110 and 126, preferably any one of SEQ ID NOs: 20 to 24, 94, 99 or 102-107; and(b) the VLP core comprises the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent.

39. A VLP according to any one of claims 31 to 38, which has a diameter of less than about 100 nm or less than about 75 nm, preferably the VLP has a diameter of between about 30 nm to about 55 nm, optionally as measured by dynamic light scattering (DLS) and / or electron microscopy.

40. A nucleic acid encoding a fusion protein as defined in any one of claims 1 to 28, a chimeric PvCSP protein or immunogenic fragment thereof as defined in claims 29 or 30, a Pvs230 or immunogenic fragment thereof as defined in any one of claims 14 to 16, or a VLP as defined in any one of claims 31 to 39, wherein optionally said nucleic acid:(a) comprises a nucleic acid sequence that is at least 85% identical, at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 31, 74-82 or 116-125, preferably SEQ ID NO: 31, 120 or 125; or(b) comprises the nucleic acid sequence of SEQ ID NO: 31, 74-82 or 116-125, preferably SEQ ID NO: 31, 120 or 125.

41. A viral vector, RNA vector or DNA plasmid comprising the nucleotide sequence of claim 40, optionally operably linked to a promoter, wherein optionally the viral vector is an adenoviral vector, a vesicular stomatitis virus (VSV) vector, an adeno-associated virus (AAV) vectors, a measles virus vector, a retroviral vector, a Herpesvirus vector, an influenza virus vector or a modified Vaccinia Ankara viral vector, preferably a non-replicating adenoviral vector or a non-replicating modified Vaccinia Ankara viral vector.

42. A host cell comprising the nucleotide sequence of claim 40 or the DNA plasmid of claim 41, wherein optionally the host cell is a mammalian cell, a yeast cell or a bacterial cell.

43. A nucleic acid according to claim 40, a viral vector, RNA vector or DNA plasmid according to claim 41 or a host cell according to claim 42, which further comprises a nucleic acid sequence encoding a VLP core, and preferably encoding a VLP core with the amino acid sequence of SEQ ID NO: 30, or SEQ ID NO: 30 wherein the methionine at position 1 is absent.

44. A pharmaceutical composition comprising a fusion protein as defined in any one of claims 1 to 28, a chimeric PvCSP protein or immunogenic fragment thereof as defined in claims 29 or 30, a Pvs230 or immunogenic fragment thereof as defined in any one of claims 14 to 16, a VLP as defined in any one of claims 31 to 39, a nucleic acid as defined in claim 40 or 43 or a viral vector or DNA plasmid as defined in claim 41 or 43, and optionally one or more pharmaceutically acceptable excipient, carrier and / or diluent.

45. A vaccine composition comprising a fusion protein as defined in any one of claims 1 to 28, a chimeric PvCSP protein or immunogenic fragment thereof as defined in claims 29 or 30, a Pvs230 or immunogenic fragment thereof as defined in any one of claims 14 to 16, a VLP as defined in any one of claims 31 to 39, a nucleic acid as defined in claim 40 or 43 or a viral vector or DNA plasmid as defined in claim 41 or 43, and a pharmaceutically acceptable adjuvant.

46. A vaccine composition according to claim 45, wherein the pharmaceutically acceptable adjuvant is selected from MF59, AS01 or biosimilars thereof, or a combination thereof.

47. A pharmaceutical composition according to claim 44 or a vaccine composition according to claim 45 or 46, which is stable at 4°C, 25°C and 37°C for at least 2 weeks.

48. A pharmaceutical composition according to claim 44 or 47 or a vaccine composition according to any one of claims 45 to 47, which is stable at -80°C for at least 3 months.

49. A pharmaceutical composition according to claim 44, 47 or 48 or a vaccine composition according to any one of claims 45 to 48, which is a liquid or solid formulation.

50. A vaccine composition according to any one of claims 45 to 49, which induces antibodies with a geometric mean titre (GMT) of:(a) at least 50,000 lU / mL, optionally at least 60,000 lU / mL against PvCSP; and / or(b) at least 5,000 ILJ / mL, optionally at least 6,000 lU / mL, at least 7,000 lU / mL or at least 8,000 lU / mL against Pvs230; and / or(c) at least 50,000 lU / mL, optionally at least 60,000 lU / mL against PVDBPIL51. A pharmaceutical preparation comprising:(a) a pharmaceutical composition according to claim 44, 47, 48 or 49 or a vaccine composition according to any one of claims 45 to 50; and(b) (i) one or more additional P. vivax polypeptide, or a variant, derivative or fragment thereof, or a nucleic acid encoding said polypeptide; and / or (ii) one or more P. falciparum polypeptide, or a variant, derivative or fragment thereof, or a nucleic acid encoding said polypeptide; as a combined preparation in a form suitable for simultaneous, separate or sequential use.

52. A method of producing a fusion protein as defined in any one of claims 1 to 28, a chimeric PvCSP protein or immunogenic fragment thereof as defined in claims 29 or 30, a Pvs230 or immunogenic fragment thereof as defined in any one of claims 14 to 16, or a VLP as defined in any one of claims 31 to 39, the method comprising culturing a host cell as defined in claim 42 under conditions suitable to express said fusion protein, chimeric PvCSP protein or immunogenic fragment thereof, Pvs230 or immunogenic fragment thereof, or VLP, and recovering the fusion protein, chimeric PvCSP protein or immunogenic fragment thereof, Pvs230 or immunogenic fragment thereof, or VLP produced, optionally wherein said method further comprises:(a) introducing a nucleic acid as defined in claim 40 into said host cell; and / or(b) purifying the fusion protein, chimeric PvCSP protein or immunogenic fragment thereof, Pvs230 or immunogenic fragment thereof, or VLP produced.

53. A fusion protein as defined in any one of claims 1 to 28, a chimeric PvCSP protein or immunogenic fragment thereof as defined in claims 29 or 30, a Pvs230 or immunogenic fragment thereof as defined in any one of claims 14 to 16, a VLP as defined in any one of claims 31 to 39, a nucleic acid as defined in claim 40 or 43, a viral vector, RNA vector or DNA plasmid as defined in claim 41 or 42, a pharmaceutical composition as defined in claim 44, 47, 48 or 49, a vaccine composition as defined in any one of claims 45 to 50, or a pharmaceutical preparation as defined in claim 51 for use in a method of preventing and / or treating malaria.

54. A method of preventing and / or treating malaria comprising the administration of an effective dose of fusion protein as defined in any one of claims 1 to 28, a chimeric PvCSP protein or immunogenicfragment thereof as defined in claims 29 or 30, a Pvs230 or immunogenic fragment thereof as defined in any one of claims 14 to 16, a VLP as defined in any one of claims 31 to 39, a nucleic acid as defined in claim 40 or 43, a viral vector, RNA vector or DNA plasmid as defined in claim 41 or 42, a pharmaceutical composition as defined in claim 44, 47, 48 or 49, a vaccine composition as defined in any one of claims 45 to 50, or a pharmaceutical preparation as defined in claim 51 to a patient in need thereof.

55. Use of a fusion protein as defined in any one of claims 1 to 28, a chimeric PvCSP protein or immunogenic fragment thereof as defined in claims 29 or 30, a Pvs230 or immunogenic fragment thereof as defined in any one of claims 14 to 16, a VLP as defined in any one of claims 31 to 39, a nucleic acid as defined in claim 40 or 43, or a viral vector or DNA plasmid as defined in claim 41 or 42, in the manufacture of a medicament for the prevention and / or treatment of malaria.

Images