Compositions for delivery of plasmodium RH5 antigens and related methods

Abstract

The present disclosure provides compositions (e.g., pharmaceutical compositions) for delivery of malarial protein antigens and related technologies (e.g., components thereof and / or methods relating thereto). Among other things, the present disclosure provides polyribonucleotides encoding malarial protein antigens.

Description

COMPOSITIONS FOR DELIVERY OF PLASMODIUM RH5 ANTIGENS AND RELATED METHODS BACKGROUND

[0001] Malaria is a mosquito-borne infectious disease caused by protozoan parasites of the Plasmodium genus. According to the World Health Organization, an estimated 3.4 billion people in 92 countries are at risk of being infected with the malaria parasite and developing disease.SUMMARY

[0002] The present disclosure provides technologies (e.g., compositions, methods, etc.) for delivery of Plasmodium antigens (also referred to herein as "malaria antigens"). In one aspect, provided herein is a polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises one or more Plasmodium Rh5 polypeptides or antigenic portions thereof. In some embodiments, a portion is an antigenic portion.

[0003] In some embodiments, a polyribonucleotide as provided herein is an isolated polyribonucleotide. In some embodiments, a polyribonucleotide as provided herein is an engineered polyribonucleotide. In some embodiments, a polyribonucleotide as provided herein is a codon-optimized polyribonucleotide.

[0004] In some embodiments, a polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises one or more Plasmodium reticulocyte-binding protein homolog 5 (Rh5) polypeptides or antigenic portions thereof. In some embodiments, one or more Plasmodium Rh5 polypeptides or antigenic portions thereof comprise one or more antigenic portions of Plasmodium Rh5.

[0005] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise one or more ordered domains of Rh5. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an N-terminal ordered domain. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 24. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 26. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 22. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 20. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 31.

[0006] In some embodiments, an N-terminal ordered domain comprises one or more N-linked glycosylation sites. In some embodiments, an N-terminal ordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites.

[0007] In some embodiments, an N-terminal ordered domain comprises a NX[T / S] to QX[T / S] amino acid substitution at one or more N-linked glycosylation sites. In some embodiments, an N-terminal ordered domain comprises a NX[T / S] to NXA amino acid substitution at one or more N-linked glycosylation sites. In some embodiments, an N-terminal ordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises: (i) NX[T / S] to QX[T / S], or (ii) NX[T / S] to NXA. In some embodiments, an N-terminal ordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises T216A, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, an N-terminal ordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises N214Q, wherein the amino acid numbering is relative to SEQ ID NO: 1.

[0008] In some embodiments, an N-terminal ordered domain comprises a C203Y amino acid substitution. In some embodiments, an N-terminal ordered domain comprises a U57L amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, an N-terminal ordered domain comprises a D183E amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, an N-terminal ordered domain comprises a A233K amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, an N-terminal ordered domain comprises: (i) I157L, (ii) D183E, (iii) A233K, or (iv) any combination thereof, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments,an N-terminal ordered domain comprises: (i) U57L, (ii) D183E, and (iii) A233K, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, an N-terminal ordered domain comprises a L231T amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, an N-terminal ordered domain comprises: (i) D226N, (ii) K228T, (iii) K237N, (iv) L238L, (v) E239T or (vi) any combination thereof, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, an N-terminal ordered domain comprises: i) D226N, (ii) K228T, (iii) K237N, (iv) L238L, and (v) E239T, wherein the amino acid numbering is relative to SEQ ID NO: 1.

[0009] In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 28. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 29. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 19. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 21. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 23. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 25. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 7. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 30. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 32. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 33. In some embodiments, an N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 34.

[0010] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a C-terminal ordered domain. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 36. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 45. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 39.

[0011] In some embodiments, a C-terminal ordered domain comprises one or more N-linked glycosylation sites. In some embodiments, a C-terminal ordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites. In some embodiments, a C-terminal ordered domain comprises a NX[T / S] to QX[T / S] amino acid substitution at one or more N-linked glycosylation sites. In some embodiments, a C-terminal ordered domain comprises a NX[T / S] to NXA amino acid substitution at one or more N-linked glycosylation sites. In some embodiments, a C-terminal ordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises: (i) NX[T / S] to QX[T / S], or (ii) NX[T / S] to NXA.

[0012] In some embodiments, a C-terminal ordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises T299A, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises N297Q, wherein the amino acid numbering is relative to SEQ ID NO: 1.

[0013] In some embodiments, a C-terminal ordered domain comprises a M304F amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1.

[0014] In some embodiments, a C-terminal ordered domain comprises a K312N amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a L314F amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a K316N amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a M330N amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a S370A amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In someembodiments, a C-terminal ordered domain comprises a S381N amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a T384K amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a L387T amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a N298T amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a L392K amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a T395N amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a N398E amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a R458K amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a N464K amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a S467A amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises a F505L amino acid substitution, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises: (i) M304F, (ii) K312N, (iii) L314F, (iv) K316N, (v) M330N, (vi) S370A, (vii) S381N, (viii) T384K, (ix) L392K, (x) T395N, (xi) N398E, (xii) R458K, (xiii) N464K, (xiv) S467A, (xv) F505L, or (xvi) any combination thereof, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises: (i) M304F, (ii) K312N, (iii) L314F, (iv) K316N, (v) M330N, (vi) S370A, (vii) S381N, (viii) T384K, (ix) L392K, (x) T395N, (xi) N398E, (xii) R458K, (xiii) N464K, (xiv) S467A, and (xv) F505L, wherein the amino acid numbering is relative to SEQ ID NO: 1.

[0015] In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 38. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 37. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 35. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 44. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 40. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 41. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 42. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 43. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 148. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 149. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 150. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 152. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 153. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 154. In some embodiments, a C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 155.

[0016] In some embodiments, one or more antigenic portions of Plasmodium Rh5 comprise a linking disordered domain. In some embodiments, a linking disordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 49.

[0017] In some embodiments, a linking disordered domain comprises one or more N-linked glycosylation sites. In some embodiments, a linking disordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites. In some embodiments, a linking disordered domain comprises an amino acid substitution at one ormore N-linked glycosylation sites that comprises NX[T / S] to QX[T / S], In some embodiments, a linking disordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises NX[T / S] to NXA. In some embodiments, a linking disordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises: (i) NX[T / S] to QX[T / S], or (ii) NX[T / S] to NXA.

[0018] In some embodiments, a linking disordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises T286A, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a linking disordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises N284Q, wherein the amino acid numbering is relative to SEQ ID NO: 1.

[0019] In some embodiments, a linking disordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 48. In some embodiments, a linking disordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 46.

[0020] In some embodiments, one or more antigenic portions of Plasmodium Rh5 comprise an N-terminal disordered domain. In some embodiments, an N-terminal disordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 52.

[0021] In some embodiments, an N-terminal disordered domain comprises one or more N-linked glycosylation sites. In some embodiments, an N-terminal disordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites.

[0022] In some embodiments, an N-terminal disordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises NX[T / S] to QX[T / S], In some embodiments, an N-terminal disordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises NX[T / S] to NXA. In some embodiments, an N-terminal disordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises: (i) NX[T / S] to QX[T / S], or (ii) NX[T / S] to NXA. In some embodiments, an N-terminal disordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises T40A, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, an N-terminal disordered domain comprises an amino acid substitution at one or more N-linked glycosylation sites that comprises N38Q, wherein the amino acid numbering is relative to SEQ ID NO: 1.

[0023] In some embodiments, an N-terminal disordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 50. In some embodiments, a linking disordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 51.

[0024] In some embodiments, a polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises one or more linkers. In some embodiments, one or more linkers comprises a linker between the N-terminal ordered domain and the C-terminal ordered domain. In some embodiments, a linker is a glycine-serine linker. In some embodiments, a linker comprises an amino acid sequence according to SEQ ID NO: 107.

[0025] In some embodiments, a polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises: (i) an N-terminal ordered domain, and (ii) a C-terminal ordered domain. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 8. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 11. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 14. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 15. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 12. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 13. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 138. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 141. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 143. In some embodiments, a polypeptidecomprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 144. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 142.

[0026] In some embodiments, a polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises: (i) an N-terminal ordered domain, (ii) a linker disordered domain, and (iii) a C-terminal ordered domain. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 2. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 3. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 4. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 5. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 109. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 110. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 132. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 133. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 134. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 135.

[0027] In some embodiments, a polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises: (i) an N-terminal ordered domain, (ii) a linker, (iii) a C-terminal ordered domain. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 6. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 7. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 9. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 10. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 136. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 137. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 139. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 140.

[0028] In some embodiments, a polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises: (i) an N-terminal disordered domain, (ii) an N-terminal ordered domain, and (iii) a C-terminal ordered domain. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 16. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 17. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 18. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 145. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 146. In some embodiments, a polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 147.

[0029] In some embodiments, a polyribonucleotide encodes a polypeptide, wherein the polypeptide comprises a secretory signal. In some embodiments, a secretory signal comprises or consists a Plasmodium secretory signal. In some embodiments, a secretory signal comprises or consists of a heterologous secretory signal. In some embodiments, a heterologous secretory signal comprises or consists of a non-human secretory signal. In some embodiments, a heterologous secretory signal comprises or consists of a viral secretory signal. In some embodiments, a viral secretory signal comprises or consists of an HSV secretory signal. In some embodiments, an HSV secretory signal comprises or consists of an HSV-1 or HSV-2 secretory signal. In some embodiments, an HSV secretory signal comprises or consists of an HSV glycoprotein D (gD) secretory signal. In some embodiments, an HSV gD secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 58. In some embodiments, a secretory signal is located at the N-terminus of the polypeptide.

[0030] In some embodiments, an RNA construct comprises, in 5' to 3' order: (i) a 5‘ UTR; (ii) a polyribonucleotide described herein; (iii) a 3‘ UTR; and (iv) a polyA tail sequence.

[0031] The present disclosure also provides compositions. In some embodiments, a composition comprises one or more polyribonucleotides provided herein. In some embodiments, a composition comprises one or more RNA constructs provided herein.

[0032] In some embodiments, a composition comprises (i) one or more polyribonucleotides and (ii) lipid nanoparticles, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes. In some embodiments, the one or more polyribonucleotides are fully or partially encapsulated within the lipid nanoparticles, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes.

[0033] In some embodiments, a composition comprises (i) one or more polyribonucleotides and (ii) lipid nanoparticles. In some embodiments, the one or more polyribonucleotides are fully or partially encapsulated within the lipid nanoparticles. In some embodiments, lipid nanoparticles each comprise: (a) a polymer-conjugated lipid; (b) a cationic lipid; and (c) one or more neutral lipids.

[0034] The present disclosure provides pharmaceutical compositions. In some embodiments, a pharmaceutical composition comprises (i) one or more polyribonucleotides as described herein, one or more RNA constructs as described herein, or a composition as described herein, and (ii) at least one pharmaceutically acceptable excipient.

[0035] The present disclosure provides, among other things, combinations of one or more pharmaceutical compositions. In some embodiments, a combination comprises: (i) a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide is a polyribonucleotide provided herein; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium Rh5 invasion complex polypeptides selected from: one or more CyRPA polypeptides or antigenic portions thereof, one or more Ripr polypeptides or antigenic portions thereof, one or more Pl 13 polypeptides or antigenic portions thereof, one or more TRAMP polypeptides or antigenic portions thereof, one or more CSS polypeptides or antigenic portions thereof, or a combination thereof.

[0036] In some embodiments, a combination comprises: (i) a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide is a polyribonucleotide provided herein; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium T cell antigens.

[0037] In some embodiments, a combination comprises: (i) a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide is a polyribonucleotide provided herein; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium CSP polypeptides or antigenic portions thereof. In some embodiments, a combination comprises (i) a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide is a polyribonucleotide according to any one of claims 1-10; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium T cell antigens, optionally selected from Plasmodium CSP, Pfs230, LISP-1, LISP-2, TRAP, LSAP2, LSA-3, LSA-l(a), LSA-l(b), UIS3, ETRAMP10.3, TLP, CelTos, PLP1, SPECT1, SPECT2, STARP, MAEBL, CLAMP, SALSA, SIAP1, LIMP, GEST, CyRPA, CSS, Ripr, TRAMP, GARP, PIESP2 and P113.

[0038] Among other things, the present disclosure provides a method comprising administering a polyribonucleotide as described herein to a subject. The present disclosure also provides a method comprising administering an RNA construct described herein to a subject. The present disclosure further provides a method comprising administering a composition described herein to a subject. Additionally, the present disclosure provides a method comprising administering one or more doses of the pharmaceutical composition described herein to a subject.The present disclosure also provides a method comprising administering a combination of one or more pharmaceutical compositions described herein to a subject.

[0039] In some embodiments, a polyribonucleotide provided herein is for use in the treatment of a malaria infection comprising administering one or more doses of the pharmaceutical composition to a subject.

[0040] In some embodiments, a composition provided herein is for use in the treatment of a malaria infection comprising administering one or more doses of the pharmaceutical composition to a subject.

[0041] In some embodiments, a pharmaceutical composition as provided herein is for use in the treatment of a malaria infection comprising administering one or more doses of the pharmaceutical composition to a subject.

[0042] In some embodiments, a combination of one or more pharmaceutical compositions provided herein is for use in the treatment of a malaria infection comprising administering one or more doses of the pharmaceutical composition to a subject.

[0043] In some embodiments, a polyribonucleotide provided herein is for use in the prevention of a malaria infection comprising administering one or more doses of the pharmaceutical composition to a subject.

[0044] In some embodiments, a composition provided herein is for use in the prevention of a malaria infection comprising administering one or more doses of the pharmaceutical composition to a subject.

[0045] In some embodiments, a pharmaceutical composition as provided herein is for use in the prevention of a malaria infection comprising administering one or more doses of the pharmaceutical composition to a subject.

[0046] In some embodiments, a combination of one or more pharmaceutical compositions provided herein is for use in the prevention of a malaria infection comprising administering one or more doses of the pharmaceutical composition to a subject.

[0047] In some embodiments, a polyribonucleotide provided herein is for use in the treatment of a malaria infection. In some embodiments, a composition provided herein is for use in the treatment of a malaria infection. In some embodiments, a pharmaceutical composition as provided herein is for use in the treatment of a malaria infection. In some embodiments, a combination of one or more pharmaceutical compositions provided herein is for use in the treatment of a malaria infection.

[0048] In some embodiments, a polyribonucleotide provided herein is for use in the prevention of a malaria infection. In some embodiments, a composition provided herein is for use in the prevention of a malaria infection. In some embodiments, a pharmaceutical composition as provided herein is for use in the prevention of a malaria infection. In some embodiments, a combination of one or more pharmaceutical compositions provided herein is for use in the prevention of a malaria infection.

[0049] In some embodiments, a polyribonucleotide provided herein is for use in inducing an anti-malaria immune response in a subject. In some embodiments, a composition provided herein is for use in inducing an anti-malaria immune response in a subject. In some embodiments, a pharmaceutical composition as provided herein is for use in inducing an anti-malaria immune response in a subject. In some embodiments, a combination of one or more pharmaceutical compositions provided herein is for use in inducing an anti-malaria immune response in a subject.

[0050] In some embodiments, a method is a method of treating a malaria infection.

[0051] In some embodiments, a method is a method of preventing a malaria infection.

[0052] Additionally, the present disclosure provides a polypeptide encoded by a polyribonucleotide described herein or an RNA construct described herein.

[0053] The present disclosure also provides a host cell comprising a polyribonucleotide described herein or an RNA construct described herein. The present disclosure further provides a host cell comprising a polypeptide described herein.BRIEF DESCRIPTION OF THE DRAWINGS

[0054] FIG. 1, parts (A) through (Q), includes schematics of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino acid (aa) references included refer to the P. falciparum Rh5 sequence set out in SEQ ID NO: 1. (A) includes a schematic of an exemplary polypeptide, RNA Construct 1.0, encoded by apolyri bonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 132. RNA Construct 1.0 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 145-247), a linking " L" disordered region (aa 248-296), and a C-terminal ordered region (297-526). Three glycosylation sites at positions 216, 286, and 299 have been mutated from N-X-[T / S] to N-X-A (where X is not proline). (B) includes a schematic of an exemplary polypeptide, RNA Construct 1.1, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 133. RNA Construct 1.1 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 145-247) comprising a C203Y mutation, a linking " L" disordered region (aa 248-296), and a C-terminal ordered region (297-526). Three glycosylation sites at positions 216, 286, and 299 have been mutated from N-X-[T / S] to N-X-A (where X is not proline). (C) includes a schematic of an exemplary polypeptide, RNA Construct 1.2, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 134. RNA Construct 1.2 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 145-247) comprising a C203Y mutation, a linking " L" disordered region (aa 248-296), and a C-terminal ordered region (297-526). Three glycosylation sites at positions 214, 284, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline). (D) includes a schematic of an exemplary polypeptide, RNA Construct 2.0, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 135. RNA Construct 2.0 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 145-247) comprising a C203Y mutation, a linking " L" disordered region (aa 248-296), and a C-terminal ordered region (297-506). Three glycosylation sites at positions 214, 284, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline). (E) includes a schematic of an exemplary polypeptide, RNA Construct 3.0, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 136. RNA Construct 3.0 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 145-247) comprising a C203Y mutation, a glycine-serine linker (e.g., SEQ ID NO: 107), and a C-terminal ordered region (297-506). Two glycosylation sites at positions 214 and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline). (F) includes a schematic of an exemplary polypeptide, RNA Construct 3.1, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 137. RNA Construct 3.1 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 145-240) comprising a C203Y mutation, a glycine-serine linker (e.g., 107), and a C-terminal ordered region (297-506). Two glycosylation sites at positions 214, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline). (G) includes a schematic of an exemplary polypeptide, RNA Construct 3.2, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 138. RNA Construct 3.2 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 145-247) comprising a C203Y mutation, and a C-terminal ordered region (297-506). Two glycosylation sites at positions 214, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline). (H) includes a schematic of an exemplary polypeptide, RNA Construct 3.3, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 139. RNA Construct 3.3 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 145-240) comprising a C203Y mutation, a glycine-serine linker (e.g., SEQ ID NO: 107), and a C-terminal ordered region (297-497). Two glycosylation sites at positions 214, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline). (I) includes a schematic of an exemplary polypeptide, RNA Construct 3.4, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 140. RNA Construct 3.4 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 155-247) comprising a C203Y mutation, a glycine-serine linker (e.g., SEQ ID NO: 107), and a C-terminal ordered region (297-497). Two glycosylation sites at positions 214, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline). (J) includes a schematic of an exemplary polypeptide, RNA Construct 4.0, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 141. RNA Construct 4.0 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal orderedregion (aa 155-247) comprising a C203Y mutation, and a C-terminal ordered region (297-506). Two glycosylation sites at positions 214, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline). (K) includes a schematic of an exemplary polypeptide, RNA Construct 5.0, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 142. RNA Construct 5.0 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 155-247) comprising a C203Y mutation, and a C-terminal ordered region (297-497). Two glycosylation sites at positions 214, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline). (L) includes a schematic of an exemplary polypeptide, RNA Construct 6.0, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 13. RNA Construct 6.0 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 145-247), and a C-terminal ordered region (297-506). Two glycosylation sites at positions 216, and 299 have been mutated from N-X-[T / S] to N-X-A (where X is not proline). (M) includes a schematic of an exemplary polypeptide, RNA Construct 7.0, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 143. RNA Construct 7.0 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 140-247) comprising a C203Y mutation, and a C-terminal ordered region (297-506). Two glycosylation sites at positions 214, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline), and 3 glycosylation sites have been created by L231T, L387T, and N398T mutations. (N) includes a schematic of an exemplary polypeptide, RNA Construct 7.1, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 144. RNA Construct 7.1 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a N-terminal ordered region (aa 140-247) comprising a C203Y mutation, and a C-terminal ordered region (297-506). Two glycosylation sites at positions 214, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline), and 6 glycosylation sites have been created by K228T, E239T, M304T, L387T, N398T, and L497T mutations. (O) includes a schematic of an exemplary polypeptide, RNA Construct 8.0, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 145. RNA Construct 8.0 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a linking N-terminal " N" disordered region (aa 26-139), an N-terminal ordered region (aa 140-247), and a C-terminal ordered region (297-526). Three glycosylation sites at positions 40, 216, and 299 have been mutated from N-X-[T / S] to N-X-A (where X is not proline). (P) includes a schematic of an exemplary polypeptide, RNA Construct 8.1, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 146. RNA Construct 8.1 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a linking N-terminal " N" disordered region (aa 26-139), an N-terminal ordered region (aa 140-247), and a C-terminal ordered region (297-506). Three glycosylation sites at positions 40, 216, and 299 have been mutated from N-X-[T / S] to N-X-A (where X is not proline). (Q) includes a schematic of an exemplary polypeptide, RNA Construct 8.2, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 147. RNA Construct 8.2 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), a linking N-terminal " N" disordered region (aa 26-139), an N-terminal ordered region (aa 140-247) comprising a C203Y mutation, and a C-terminal ordered region (297-506). Three glycosylation sites at positions 38, 214, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline). (R) includes a schematic of an exemplary polypeptide, RNA Construct 2.1, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 109. RNA Construct 2.1 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), an additional Aspartic acid between the signal peptide and the N-terminal sequence, a N-terminal ordered region (aa 145-247) comprising a C203Y mutation, a linking " L" disordered region (aa 248-296), and a C-terminal ordered region (297-506). Three glycosylation sites at positions 214, 284, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline). (S) includes a schematic of an exemplary polypeptide, RNA Construct 2.2, encoded by a polyribonucleotide described herein and having the amino acid sequence according to SEQ ID NO: 110. RNA Construct 2.2 comprises an HSV glycoprotein D (gD) secretory signal (e.g., SEQ ID NO: 58), an additional 2 Aspartic acids between the signal peptide and the N-terminal sequence, a N-terminal ordered region (aa 145-247) comprising a C203Y mutation, a linking " L" disordered region (aa 248-296), anda C-terminal ordered region (297-506). Three glycosylation sites at positions 214, 284, and 297 have been mutated from N-X-[T / S] to Q-X-[T / S] (where X is not proline).

[0055] Fig. 2 depicts in vitro expression of non-formulated RNA constructs in HEK293T cells using MessengerMax as transfection reagent. FIG. 2A shows the transfection rate as measured by percentage of HEK293T population that is positive for the presence of expressed protein, providing a measure of efficiency of RNA delivery into the cell. FIG.2B shows the total expression as measured by median fluorescence of the total HEK293T population for both transfected and non-transfected cells. Total expression of permeabilized cells shows total protein expressed.

[0056] Fig. 3 depicts ELISA OD values and endpoint antibodies titers against RH5 for serum samples collected on Day 21 (Fig. 3A / B) (pre-boost) and Day 35 (Fig. 3C / D) (post-boost). Briefly, the RNA constructs were assessed for their ability to induce antibodies that bind to Plasmodium falciparum P) RH5 protein (recombinantly expressed RH5 protein with unstructured regions removed, using an ELISA assay.

[0057] Provided polyribonucleotides were assessed for their ability to induce production of antibodies that inhibit parasite blood stage growth in a growth inhibition assay (GIA). Total IgG was purified from sera collected from mice immunized twice with exemplary polyribonucleotide constructs. The viability of blood stage parasites was determined after 42 hours of culture in the presence of total IgG from immunized mice (ranging from 2 - 0.015625 mg / mL) (Fig.4A to O).DEFINITIONS

[0058] Compounds of this disclosure include those described generally above and are further illustrated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this disclosure, the chemical elements are identified in accordance with the Periodic Table of Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed. Additionally, general principles of organic chemistry are described in " Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and " March's Advanced Organic Chemistry", 5th Ed., Ed.: Smith, M. B. and March, J., John Wiley & Sons, New York: 2001, the entire contents each of which are hereby incorporated by reference.

[0059] Unless otherwise stated, structures depicted herein are meant to include all stereoisomeric (e.g., enantiomeric or diastereomeric) forms of the structure, as well as all geometric or conformational isomeric forms of the structure. For example, the R and S configurations of each stereocenter are contemplated as part of the disclosure. Therefore, single stereochemical isomers, as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of provided compounds are within the scope of the disclosure. For example, in some cases, provided compounds show one or more stereoisomers of a compound, and unless otherwise indicated, represents each stereoisomer alone and / or as a mixture. Unless otherwise stated, all tautomeric forms of provided compounds are within the scope of the disclosure.

[0060] Unless otherwise indicated, structures depicted herein are meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures including replacement of hydrogen by deuterium or tritium, or replacement of a carbon by 13C- or 14C-enriched carbon are within the scope of this disclosure.

[0061] About. The term "about," when used herein in reference to a value, refers to a value that is similar, in context to the referenced value. In general, those skilled in the art, familiar with the context, will appreciate the relevant degree of variance encompassed by "about" in that context. For example, in some embodiments, the term "about" may encompass a range of values that within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value.

[0062] Agent. As used herein, the term "agent" may refer to a physical entity. In some embodiments, an agent may be characterized by a particular feature and / or effect. For example, as used herein, the term "therapeutic agent" refers to a physical entity has a therapeutic effect and / or elicits a desired biological and / or pharmacological effect. Insome embodiments, an agent may be a compound, molecule, or entity of any chemical class including, for example, a small molecule, polypeptide, nucleic acid, saccharide, lipid, metal, or a combination or complex thereof.

[0063] Amino acid. In its broadest sense, as used herein, the term "amino acid" refers to a compound and / or substance that can be, is, or has been incorporated into a polypeptide chain, e.g., through formation of one or more peptide bonds. In some embodiments, an amino acid has the general structure H2N-C(H)(R)-COOH. In some embodiments, an amino acid is a naturally-occurring amino acid. In some embodiments, an amino acid is a non-natural amino acid; in some embodiments, an amino acid is a D-amino acid; in some embodiments, an amino acid is an L-amino acid. " Standard amino acid" refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. " Nonstandard amino acid" refers to any amino acid, other than the standard amino acids, regardless of whether it is prepared synthetically or obtained from a natural source. In some embodiments, an amino acid, including a carboxy- and / or amino-terminal amino acid in a polypeptide, can contain a structural modification as compared with the general structure above. For example, in some embodiments, an amino acid may be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and / or substitution (e.g., of the amino group, the carboxylic acid group, one or more protons, and / or the hydroxyl group) as compared with the general structure. In some embodiments, such modification may, for example, alter the circulating half-life of a polypeptide containing the modified amino acid as compared with one containing an otherwise identical unmodified amino acid. In some embodiments, such modification does not significantly alter a relevant activity of a polypeptide containing the modified amino acid, as compared with one containing an otherwise identical unmodified amino acid. As will be clear from context, in some embodiments, the term "amino acid" may be used to refer to a free amino acid; in some embodiments it may be used to refer to an amino acid residue of a polypeptide.

[0064] Antigen: The term "antigen," as used herein, refers to an agent that (i) elicits an immune response; and / or (ii) an agent that binds to a T cell receptor e.g., when presented by an MHC molecule) or to an antibody.

[0065] Anti-malaria immune response. The term "anti-malaria immune response," as used herein, refers to an immune response directed to one or more antigens derived from Plasmodium.

[0066] Associated. Two events or entities are "associated" with one another, as that term is used herein, if the presence, level, degree, type and / or form of one is correlated with that of the other. For example, a particular entity (e.g., polypeptide, genetic signature, metabolite, microbe, etc.) is considered to be associated with a particular disease, disorder, or condition, if its presence, level and / or form correlates with incidence of, susceptibility to, severity of, stage of, etc. the disease, disorder, or condition (e.g., across a relevant population). In some embodiments, two or more entities are physically "associated" with one another if they interact, directly or indirectly, so that they are and / or remain in physical proximity with one another. In some embodiments, two or more entities that are physically associated with one another are covalently linked to one another; in some embodiments, two or more entities that are physically associated with one another are not covalently linked to one another but are non-covalently associated, for example by means of hydrogen bonds, van der Waals interaction, hydrophobic interactions, magnetism, and combinations thereof.

[0067] Characteristic portion-. As used herein, the term "characteristic portion," in the broadest sense, refers to a portion of a polypeptide or region thereof whose presence (or absence) correlates with presence (or absence) of a particular feature, attribute, or activity of the polypeptide or region thereof. In some embodiments, a characteristic portion of a polypeptide or region thereof is a portion that is found in the polypeptide or region thereof and in related polypeptide or region thereof that share the particular feature, attribute or activity, but not in those that do not share the particular feature, attribute or activity. In certain embodiments, a characteristic portion shares at least one functional characteristic with the intact polypeptide or region thereof. For example, in some embodiments, a "characteristic portion" of a polypeptide or region thereof is one that contains a continuous stretch of amino acids, or a collection of continuous stretches of amino acids, that together are characteristic of the polypeptide or region thereof. In some embodiments, each such continuous stretch generally contains at least 2, 5, 10, 15, 20, 50, or more amino acids. In general, a characteristic portion of a polypeptide or region thereof is one that, in addition to the sequenceand / or structural identity specified above, shares at least one functional characteristic with the relevant intact polypeptide or region thereof. In some embodiments, a characteristic portion may be biologically active. In some embodiments, a fragment as described herein can be a portion. Accordingly, in some embodiments, a characteristic fragment can be a "characteristic portion."

[0068] Combination therapy. As used herein, the term "combination therapy" refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents (e.g., two or more antibody agents)). In some embodiments, the two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all "doses" of a first regimen are administered prior to administration of any doses of a second regimen); in some embodiments, such agents are administered in overlapping dosing regimens. In some embodiments, administration of combination therapy may involve administration of one or more agent(s) or modality(ies) to a subject receiving the other agent(s) or modality(ies) in the combination. For clarity, combination therapy does not require that individual agents be administered together in a single composition (or even necessarily at the same time), although in some embodiments, two or more agents, or active moieties thereof, may be administered together in a combination composition.

[0069] Comparable. As used herein, the term "comparable" refers to two or more agents, entities, situations, sets of conditions, etc., that may not be identical to one another but that are sufficiently similar to permit comparison there between so that one skilled in the art will appreciate that conclusions may reasonably be drawn based on differences or similarities observed. In some embodiments, comparable sets of conditions, circumstances, individuals, or populations are characterized by a plurality of substantially identical features and one or a small number of varied features. Those of ordinary skill in the art will understand, in context, what degree of identity is required in any given circumstance for two or more such agents, entities, situations, sets of conditions, etc. to be considered comparable. For example, those of ordinary skill in the art will appreciate that sets of circumstances, individuals, or populations are comparable to one another when characterized by a sufficient number and type of substantially identical features to warrant a reasonable conclusion that differences in results obtained or phenomena observed under or with different sets of circumstances, individuals, or populations are caused by or indicative of the variation in those features that are varied.

[0070] Corresponding to-. As used herein, the term "corresponding to" refers to a relationship between two or more entities. For example, the term "corresponding to" may be used to designate the position / identity of a structural element in a compound or composition relative to another compound or composition (e.g., to an appropriate reference compound or composition). For example, in some embodiments, a monomeric residue in a polymer (e.g., an amino acid residue in a polypeptide or a nucleic acid residue in a polynucleotide) may be identified as "corresponding to" a residue in an appropriate reference polymer. For example, those of ordinary skill will appreciate that, for purposes of simplicity, residues in a polypeptide are often designated using a canonical numbering system based on a reference related polypeptide, so that an amino acid "corresponding to" a residue at position 190, for example, need not actually be the 190thamino acid in a particular amino acid chain but rather corresponds to the residue found at 190 in the reference polypeptide; those of ordinary skill in the art readily appreciate how to identify "corresponding" amino acids. For example, those skilled in the art will be aware of various sequence alignment strategies, including software programs such as, for example, BLAST, CS-BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH / GLSEARCH, Genoogle, HMMER, HHpred / HHsearch, IDF, Infernal, KLAST, USEARCH, parasail, PSI-BLAST, PSI-Search, ScalaBLAST, Sequilab, SAM, SSEARCH, SWAPHI, SWAPHI-LS, SWIMM, or SWIPE that can be utilized, for example, to identify "corresponding" residues in polypeptides and / or nucleic acids in accordance with the present disclosure. Those of skill in the art will also appreciate that, in some instances, the term "corresponding to" may be used to describe an event or entity that shares a relevant similarity with another event or entity (e.g., an appropriate reference event or entity). To give but one example, a gene or protein in one organism may be described as "corresponding to" a gene or protein from another organism in order to indicate, in some embodiments, that it plays an analogous role or performs an analogous functionand / or that it shows a particular degree of sequence identity or homology, or shares a particular characteristic sequence element.

[0071] Dosing regimenr. Those skilled in the art will appreciate that the term "dosing regimen" (or "therapeutic regimen") may be used to refer to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses.

[0072] Encode-. As used herein, the term "encode" or "encoding" refers to sequence information of a first molecule that guides production of a second molecule having a defined sequence of nucleotides (e.g., a polyribonucleotide) or a defined sequence of amino acids. For example, a DNA molecule can encode an RNA molecule (e.g., by a transcription process that includes a DNA-dependent RNA polymerase enzyme). An RNA molecule can encode a polypeptide (e.g., by a translation process). Thus, a gene, a cDNA, or an RNA molecule encodes a polypeptide if transcription and translation of RNA corresponding to that gene produces the polypeptide in a cell or other biological system. In some embodiments, a coding region of a polyribonucleotide encoding a target antigen refers to a coding strand, the nucleotide sequence of which is identical to the polyribonucleotide sequence of such a target antigen. In some embodiments, a coding region of a polyribonucleotide encoding a target antigen refers to a non-coding strand of such a target antigen, which may be used as a template for transcription of a gene or cDNA.

[0073] Expression.- As used herein, the term "expression" of a nucleic acid sequence refers to the generation of a gene product from the nucleic acid sequence. In some embodiments, a gene product can be a transcript, e.g., a polyribonucleotide as provided herein. In some embodiments, a gene product can be a polypeptide. In some embodiments, expression of a nucleic acid sequence involves one or more of the following: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of an RNA transcript (e.g., by splicing, editing, etc.); (3) translation of an RNA into a polypeptide or protein; and / or (4) post-translational modification of a polypeptide or protein.

[0074] Heterologous: As used herein, the term "heterologous", with respect to secretory signal or transmembrane region, refers to a secretory signal or transmembrane region from a virus or an organism other than Plasmodium.

[0075] Homology. As used herein, the term "homology" or "homolog" refers to the overall relatedness between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or between polypeptide molecules. In some embodiments, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are considered to be "homologous" to one another if their sequences are at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical. In some embodiments, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are considered to be "homologous" to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% similar (e.g., containing residues with related chemical properties at corresponding positions). For example, as is well known by those of ordinary skill in the art, certain amino acids are typically classified as similar to one another as "hydrophobic" or "hydrophilic" amino acids, and / or as having "polar" or "non-polar" side chains. Substitution of one amino acid for another of the same type may often be considered a "homologous" substitution.

[0076] Identity. As used herein, the term "identity" refers to the overall relatedness between polynucleotide molecules {e.g., DNA molecules and / or RNA molecules) and / or between polypeptide molecules. In some embodiments, polynucleotide molecules e.g., DNA molecules and / or RNA molecules) and / or between polypeptide molecules are considered to be "substantially identical" to one another if their sequences are at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical. Calculation of the percent identity of two nucleic acid or polypeptide sequences, for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second sequence for optimal alignment and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of a sequence aligned for comparisonpurposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or substantially 100% of the length of a reference sequence. The nucleotides at corresponding positions are then compared. When a position in the first sequence is occupied by the same residue (e.g., nucleotide or amino acid) as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller, 1989, which has been incorporated into the ALIGN program (version 2.0). In some exemplary embodiments, nucleic acid sequence comparisons made with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna. CMP matrix.

[0077] Increased, Induced, or Reduced. As used herein, these terms or grammatically comparable comparative terms, indicate values that are relative to a comparable reference measurement. For example, in some embodiments, an assessed value achieved with a provided composition (e.g., a pharmaceutical composition) may be "increased" relative to that obtained with a comparable reference composition. Alternatively or additionally, in some embodiments, an assessed value achieved in a subject may be "increased" relative to that obtained in the same subject under different conditions (e.g., prior to or after an event; or presence or absence of an event such as administration of a composition (e.g., a pharmaceutical composition) as described herein, or in a different, comparable subject (e.g., in a comparable subject that differs from the subject of interest in prior exposure to a condition, e.g., absence of administration of a composition (e.g., a pharmaceutical composition) as described herein.). In some embodiments, comparative terms refer to statistically relevant differences (e.g., that are of a prevalence and / or magnitude sufficient to achieve statistical relevance). Those skilled in the art will be aware, or will readily be able to determine, in a given context, a degree and / or prevalence of difference that is required or sufficient to achieve such statistical significance. In some embodiments, the term "reduced" or equivalent terms refers to a reduction in the level of an assessed value by at least 5%, at least 10%, at least 20%, at least 50%, at least 75% or higher, as compared to a comparable reference. In some embodiments, the term "reduced" or equivalent terms refers to a complete or essentially complete inhibition, i.e., a reduction to zero or essentially to zero. In some embodiments, the term "increased" or "induced" refers to an increase in the level of an assessed value by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 80%, at least 100%, at least 200%, at least 500%, or higher, as compared to a comparable reference.

[0078] In order: As used herein with reference to a polynucleotide or polyribonucleotide, "in order" refers to the order of features from 5' to 3' along the polynucleotide or polyribonucleotide. As used herein with reference to a polypeptide, "in order" refers to the order of features moving from the N -terminal -most of the features to the C-terminal-most of the features along the polypeptide. " In order" does not mean that no additional features can be present among the listed features. For example, if Features A, B, and C of a polynucleotide are described herein as being "in order, Feature A, Feature B, and Feature C," this description does not exclude, e.g., Feature D being located between Features A and B.

[0079] Isolated. The term "isolated" means altered or removed from the natural state. For example, a nucleic acid or a peptide naturally present in a living animal is not "isolated," but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural state is "isolated." An isolated nucleic acid or protein can exist in substantially purified form, or can exist in a non-native environment such as, for example, a host cell.

[0080] Linker. As used herein, the term "linker" refers to a portion of a polypeptide that connects different regions, portions, or antigens to one another.

[0081] Lipid-. As used herein, the terms "lipid" and "lipid-like material" are broadly defined as molecules which comprise one or more hydrophobic moieties or groups and optionally also one or more hydrophilic moieties or groups. Molecules comprising hydrophobic moieties and hydrophilic moieties are also typically denoted as amphiphiles.

[0082] Merozoite stage specific Plasmodium antigen: As used herein, the term "merozoite stage specific Plasmodium antigen" refers to an antigen that is expressed during the merozoite stage of the Plasmodium life cycle. In some embodiments, a merozoite stage specific Plasmodium antigen is a Rh5 invasion complex antigen. In some embodiments, a Rh5 invasion complex antigen is or comprises a Rh5 antigen.

[0083] Multimerization region.- As used herein, the term "multimerization region" refers to a region that directs assembly of multimers into a complex, where each multimer comprises a polypeptide associated with the multimerization region.

[0084] RNA lipid nanoparticle.- As used herein, the term " RNA lipid nanoparticle" refers to a nanoparticle comprising at least one lipid and RNA molecule(s), e.g., one or more polyribonucleotides as provided herein. In some embodiments, an RNA lipid nanoparticle comprises at least one cationic amino lipid. In some embodiments, an RNA lipid nanoparticle comprises at least one cationic amino lipid, at least one helper lipid, and at least one polymer-conjugated lipid (e.g., PEG-conjugated lipid). In various embodiments, RNA lipid nanoparticles as described herein can have an average size e.g., Z-average) of about 100 nm to 1000 nm, or about 200 nm to 900 nm, or about 200 nm to 800 nm, or about 250 nm to about 700 nm. In some embodiments of the present disclosure, RNA lipid nanoparticles can have a particle size (e.g., Z-average) of about 30 nm to about 200 nm, or about 30 nm to about 150 nm, about 40 nm to about 150 nm, about 50 nm to about 150 nm, about 60 nm to about 130 nm, about 70 nm to about 110 nm, about 70 nm to about 100 nm, about 80 nm to about 100 nm, about 90 nm to about 100 nm, about 70 to about 90 nm, about 80 nm to about 90 nm, or about 70 nm to about 80 nm. In some embodiments, an average size of lipid nanoparticles is determined by measuring the average particle diameter. In some embodiments, RNA lipid nanoparticles may be prepared by mixing lipids with RNA molecules described herein.

[0085] Neutralization-. As used herein, the term "neutralization" refers to an event in which binding agents such as antibodies bind to a biological active site of a parasite such as a receptor binding protein, thereby inhibiting the parasitic infection of cells. In some embodiments, the term "neutralization" refers to an event in which binding agents eliminate or significantly reduce ability of infecting cells.

[0086] Nucleic acid / Polynucleotide.- As used herein, the term "nucleic acid" refers to a polymer of at least 10 nucleotides or more. In some embodiments, a nucleic acid is or comprises DNA. In some embodiments, a nucleic acid is or comprises RNA. In some embodiments, a nucleic acid is or comprises peptide nucleic acid (PNA). In some embodiments, a nucleic acid is or comprises a single stranded nucleic acid. In some embodiments, a nucleic acid is or comprises a double-stranded nucleic acid. In some embodiments, a nucleic acid comprises both single and doublestranded portions. In some embodiments, a nucleic acid comprises a backbone that comprises one or more phosphodiester linkages. In some embodiments, a nucleic acid comprises a backbone that comprises both phosphodiester and non-phosphodiester linkages. For example, in some embodiments, a nucleic acid may comprise a backbone that comprises one or more phosphorothioate or 5'-N-phosphoramidite linkages and / or one or more peptide bonds, e.g., as in a "peptide nucleic acid". In some embodiments, a nucleic acid comprises one or more, or all, natural residues (e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uracil). In some embodiments, a nucleic acid comprises on or more, or all, non-natural residues. In some embodiments, a non-natural residue comprises a nucleoside analog (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3 -methyl adenosine, 5-methylcytidine, C-5 propynyl -cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5 -propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 6-O-methylguanine, 2-thiocytidine, methylated bases, intercalated bases, and combinations thereof). In some embodiments, a non-natural residue comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) as compared to those in natural residues. In some embodiments, a nucleic acid has a nucleotide sequence that encodesa functional gene product such as an RNA or polypeptide. In some embodiments, a nucleic acid has a nucleotide sequence that comprises one or more introns. In some embodiments, a nucleic acid may be prepared by isolation from a natural source, enzymatic synthesis (e.g., by polymerization based on a complementary template, e.g., in vivo or in vitro, reproduction in a recombinant cell or system, or chemical synthesis. In some embodiments, a nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10.500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, 17.500, 18,000, 18,500, 19,000, 19,500, or 20,000 or more residues or nucleotides long.

[0087] Pharmaceutically effective amount. The term "pharmaceutically effective amount" or "therapeutically effective amount" refers to the amount which achieves a desired reaction or a desired effect alone or together with further doses. In the case of the treatment of a particular disease (e.g., malaria), a desired reaction in some embodiments relates to inhibition of the course of the disease (e.g., malaria). In some embodiments, such inhibition may comprise slowing down the progress of a disease (e.g., malaria) and / or interrupting or reversing the progress of the disease (e.g., malaria). In some embodiments, a desired reaction in a treatment of a disease (e.g., malaria) may be or comprise delay or prevention of the onset of a disease (e.g., malaria) or a condition (e.g., a malaria associated condition). An effective amount of a composition (e.g., a pharmaceutical composition) described herein will depend, for example, on disease (e.g., malaria) or a condition (e.g., a malaria associated condition) to be treated, the severity of such a disease (e.g., malaria) or a condition (e.g., a malaria associated condition), individual parameters of the patient, including, e.g., age, physiological condition, size and weight, the duration of treatment, the type of an accompanying therapy (if present), the specific route of administration and similar factors. Accordingly, doses of a composition (e.g., a pharmaceutical composition) described herein may depend on various of such parameters. In the case that a reaction in a patient is insufficient with an initial dose, higher doses (or effectively higher doses achieved by a different, more localized route of administration) may be used.

[0088] Polypeptide-. As used herein, the term "polypeptide" refers to a polymeric chain of amino acids. In some embodiments, a polypeptide has an amino acid sequence that occurs in nature. In some embodiments, a polypeptide has an amino acid sequence that does not occur in nature. In some embodiments, a polypeptide has an amino acid sequence that is engineered in that it is designed and / or produced through action of the hand of man. In some embodiments, a polypeptide may comprise or consist of natural amino acids, non-natural amino acids, or both. In some embodiments, a polypeptide may comprise or consist of only natural amino acids or only non-natural amino acids. In some embodiments, a polypeptide may comprise D-amino acids, L-amino acids, or both. In some embodiments, a polypeptide may comprise only D-amino acids. In some embodiments, a polypeptide may comprise only L-amino acids. In some embodiments, a polypeptide may include one or more pendant groups or other modifications, e.g., modifying or attached to one or more amino acid side chains, at the polypeptide's N-terminus, at the polypeptide's C-terminus, or any combination thereof. In some embodiments, such pendant groups or modifications comprise acetylation, amidation, lipidation, methylation, pegylation, etc., including combinations thereof. In some embodiments, a polypeptide may be cyclic, and / or may comprise a cyclic portion. In some embodiments, a polypeptide is not cyclic and / or does not comprise any cyclic portion. In some embodiments, a polypeptide is linear. In some embodiments, a polypeptide may be or comprise a stapled polypeptide. In some embodiments, the term "polypeptide" may be appended to a name of a reference polypeptide, activity, or structure; in such instances it is used herein to refer to polypeptides that share the relevant activity or structure and thus can be considered to be members of the same class or family of polypeptides. For each such class, the present specification provides and / or those skilled in the art will be aware of exemplary polypeptides within the class whose amino acid sequences and / or functions are known; in some embodiments, such exemplary polypeptides are reference polypeptides for the polypeptide class or family. In some embodiments, a member of a polypeptide class or family shows significant sequence homology or identity with, shares a common sequence motif (e.g., a characteristic sequence element) with, and / or shares a common activity (in someembodiments at a comparable level or within a designated range) with a reference polypeptide of the class; in some embodiments with all polypeptides within the class). For example, in some embodiments, a member polypeptide shows an overall degree of sequence homology or identity with a reference polypeptide that is at least about 30-40%, and is often greater than about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more and / or includes at least one region (e.g., a conserved region that may in some embodiments be or comprise a characteristic sequence element) that shows very high sequence identity, often greater than 90% or even 95%, 96%, 97%, 98%, or 99%. Such a conserved region usually encompasses at least 3-4 and often up to 35 or more amino acids; in some embodiments, a conserved region encompasses at least one stretch of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 or more contiguous amino acids. In some embodiments, a relevant polypeptide may comprise or consist of a fragment of a parent polypeptide. In some embodiments, a polypeptide is a Plasmodium polypeptide construct described herein. A Plasmodium polypeptide construct is a polypeptide that includes one or more malarial proteins, or one or more portions thereof. In some embodiments, a Plasmodium polypeptide construct described herein includes at least one Plasmodium Rh5 polypeptide or an antigenic a portion thereof. In some embodiments, a Plasmodium polypeptide construct additionally includes one or more additional amino acid sequences, such as a secretory signal (e.g., a heterologous secretory signal), a transmembrane region (e.g., a heterologous transmembrane region), a multimerization region, and / or a linker, as described herein.

[0089] Prevent. As used herein, the term "prevent" or "prevention" when used in connection with the occurrence of a disease, disorder, and / or condition, refers to reducing the risk of developing the disease, disorder and / or condition and / or to delaying onset of one or more characteristics or symptoms of the disease, disorder or condition. Prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a predefined period of time or has been avoided altogether. In some embodiments, prevention refers to reducing the risk of developing clinical malaria. In some embodiments, prevention refers to reducing the risk of developing clinical malaria by 70%, by 75%, by 80%, by 85%, by 90%, by 95%, by 98%, by 99% or by 100% in a treated population relative to an untreated population.

[0090] Reference.' As used herein, the term "reference" describes a standard or control relative to which a comparison is performed. For example, in some embodiments, an agent, animal, individual, population, sample, sequence or value of interest is compared with a reference or control agent, animal, individual, population, sample, sequence or value. In some embodiments, a reference or control is tested and / or determined substantially simultaneously with the testing or determination of interest. In some embodiments, a reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as would be understood by those skilled in the art, a reference or control is determined or characterized under comparable conditions or circumstances to those under assessment. Those skilled in the art will appreciate when sufficient similarities are present to justify reliance on and / or comparison to a particular possible reference or control.

[0091] Rh5 invasion complex-. As used herein refers to a complex that is present in Plasmodium merozoites that includes Rh5. In some embodiments, a Rh5 invasion complex facilitates merozoite binding to and / or invasion of an erythrocyte. Without limitation, polypeptides in an Rh5 invasion complex may include one or more of Plasmodium reticulocyte-binding protein homolog 5 (Rh5), P / asmodium Cyste\ne-R\ch Protective Antigen (CyRPA), Plasmodium Rh5-interacting Protein (Ripr), Plasmodium thrombospondin-related apical merozoite protein (TRAMP), and Plasmodium cysteine-rich small secreted protein (CSS). Exemplary Rh5 invasion complexes include the RCR complex and the PCRCR complex. An RCR complex is a trimeric complex comprising Rh5, cysteine-rich protective antigen (CyRPA), and Rh5-interacting protein (Ripr). A PCRCR complex is a pentameric complex comprising Rh5, CyRPA, Ripr, CyRPA, CSS and TRAMP. The protein P113 binds to and is functionally associated with the RH5 invasion complex.

[0092] Ribonucleic acid (RNA) or Polyribonucleotide-. As used herein, the term "ribonucleic acid,"" RNA," or "polyribonucleotide" refers to a polymer of ribonucleotides. In some embodiments, an RNA is single stranded. In some embodiments, an RNA is double stranded. In some embodiments, an RNA comprises both single and doublestranded portions. In some embodiments, an RNA can comprise a backbone structure as described in the definition of " Nucleic acid I Polynucleotide" above. An RNA can be a regulatory RNA (e.g., siRNA, microRNA, etc.), or a messenger RNA (mRNA). In some embodiments, an RNA is a mRNA. In some embodiments, where an RNA is a mRNA, a RNA typically comprises at its 3' end a poly(A) region. In some embodiments, where an RNA is a mRNA, an RNA typically comprises at its 5' end an art-recognized cap structure, e.g., for recognizing and attachment of a mRNA to a ribosome to initiate translation. In some embodiments, a RNA is a synthetic RNA. Synthetic RNAs include RNAs that are synthesized in vitro (e.g., by enzymatic synthesis methods and / or by chemical synthesis methods). In some embodiments, a polyribonucleotide encodes a polypeptide, which is preferably is a Plasmodium polypeptide construct.

[0093] Ribonucleotide-. As used herein, the term "ribonucleotide" encompasses unmodified ribonucleotides and modified ribonucleotides. For example, unmodified ribonucleotides include the purine bases adenine (A) and guanine (G), and the pyrimidine bases cytosine (C) and uracil (U). Modified ribonucleotides may include one or more modifications including, but not limited to, for example, (a) end modifications, e.g., 5' end modifications (e.g., phosphorylation, dephosphorylation, conjugation, inverted linkages, etc.), 3' end modifications (e.g., conjugation, inverted linkages, etc.), (b) base modifications, e.g., replacement with modified bases, stabilizing bases, destabilizing bases, or bases that base pair with an expanded repertoire of partners, or conjugated bases, (c) sugar modifications (e.g., at the 2' position or 4' position) or replacement of the sugar, and (d) internucleoside linkage modifications, including modification or replacement of the phosphodiester linkages. The term "ribonucleotide" also encompasses ribonucleotide triphosphates including modified and non-modified ribonucleotide triphosphates.

[0094] Secretory signal: As used herein, the term "secretory signal" refers to an amino acid sequence motif that targets associated polypeptides for translocation to a secretory pathway.

[0095] Subject. As used herein, the term "subject" refers to an organism to be administered with a composition described herein, e.g., for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, domestic pets, etc.) and humans. In preferred embodiments, a subject is a human subject. In some embodiments, a subject is suffering from a disease, disorder, or condition (e.g., malaria and / or a malaria-associated condition). In some embodiments, a subject is susceptible to a disease, disorder, or condition (e.g., malaria and / or a malaria-associated condition). In some embodiments, a subject displays one or more symptoms or characteristics of a disease, disorder, or condition (e.g., malaria and / or a malaria-associated condition). In some embodiments, a subject displays one or more non-specific symptoms of a disease, disorder, or condition (e.g., malaria and / or a malaria-associated condition). In some embodiments, a subject does not display any symptom or characteristic of a disease, disorder, or condition (e.g., malaria and / or a malaria-associated condition). In some embodiments, a subject is someone with one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition (e.g., malaria and / or a malaria-associated condition). In some embodiments, a subject is a patient. In some embodiments, a subject is an individual to whom diagnosis and / or therapy is and / or has been administered.

[0096] Suffering from-. An individual who is "suffering from" a disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition) has been diagnosed with and / or displays one or more symptoms of a disease, disorder, and / or condition.

[0097] Susceptible to-. An individual who is "susceptible to" a disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition) is one who has a higher risk of developing the disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition) due to their presence in an area in which malaria is endemic. In some embodiments, an individual who is susceptible to a disease, disorder and / or condition (e.g., malaria and / or a malaria-associated condition) may not have been diagnosed with the disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition). In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition) may exhibit symptoms of the disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition). In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition) may notexhibit symptoms of the disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition). In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition) will develop the disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition). In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition) will not develop the disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition).

[0098] Therapy. The term "therapy" refers to an administration or delivery of an agent or intervention that has a therapeutic effect and / or elicits a desired biological and / or pharmacological effect (e.g., has been demonstrated to be statistically likely to have such effect when administered to a relevant population). In some embodiments, a therapeutic agent or therapy is any substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition). In some embodiments, a therapeutic agent or therapy is a medical intervention that can be performed to alleviate, relieve, inhibit, present, delay onset of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition.

[0099] Transmembrane region-. As used herein, the term "transmembrane region" refers to a region of a polypeptide that spans a biological membrane, such as the plasma membrane of a cell.

[0100] Treat. As used herein, the term "treat," "treatment," or "treating" refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition). Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition). In some embodiments, treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition), for example for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and / or condition. In some embodiments, treatment may be administered to a subject at a later-stage of disease, disorder, and / or condition (e.g., malaria and / or a malaria-associated condition).

[0101] Variant: As used herein, the term "variant" refers to a molecule that shows significant structural (e.g., primary or secondary) identity with a reference molecule but differs structurally from the reference molecule. For example, a variant polypeptide or nucleic acid may differ from a reference polypeptide or nucleic acid as a result of one or more differences in amino acid or nucleotide sequence and / or one or more differences in chemical moieties {e.g., carbohydrates, lipids, phosphate groups) that are covalently components of the polypeptide or nucleic acid e.g., that are attached to the polypeptide or nucleic acid backbone).DETAILED DESCRIPTION OF CERTAIN EMBODIMENTSI. Malaria

[0102] Malaria is a mosquito-borne infectious disease caused by single-celled eukaryotic Plasmodium parasites that are transmitted by the bite of Anopheles spp. mosquitoes (Phillips, M., et al. Malaria. Nat Rev Dis Primers 3, 17050, 2017, which is incorporated herein by reference in its entirety). Mosquitoes that transmit malaria must have been infected through a previous blood meal taken from an infected subject (e.g., a human). When a mosquito bites an infected subject a small amount of blood is ingested that contains Plasmodium parasites. If the blood meal contains viable gametocytes, the Anopheles mosquito can become infected, and, upon maturation and migration of sporozoites to the salivary glands, the infectious mosquito can subsequently bite and transmit the parasites to a new host.

[0103] Malaria remains one of the most serious infectious diseases, causing approximately 200 million clinical cases and 500,000-600,000 deaths annually. Although significant effort has been invested in developing therapeutic treatments for malaria, many malaria parasites have developed resistance to available therapeutics. According to Malaria Eradication Research Agenda Initiative, malaria eradication will only be achievable through effective vaccination.

[0104] In 2015, the European Medicines Agency gave a positive review to a malaria vaccine candidate known as " RTS, S", a milestone in malaria vaccine development. In 2019, the World Health Organization launched pilot programs that provide RTS, S to children at least 5 months of age in parts of three sub-Saharan African countries. RTS, S / AS01 is an adjuvanted protein subunit vaccine that consists of a portion of the major repeat region and the C-terminus of CSP from Plasmodium falciparum fused to the Hepatitis B surface antigen (HBsAg). The vaccine is a mix of this PfCSP-HBsAg compound with HBsAg that forms virus-like particles (RTS, S / AS01; Mosquirix™). RTS, S is administered according to a regimen that requires four doses: an initial 3-dose schedule given at least 1 month apart, and a 4th dose 15-18 months after dose 3 (see, for example, Vandoolaeghe & Schuerman Expert Rev Vaccines. 15:1481, 2016; PATH_MVI_RTSS_Fact Sheet_042019, each of which is incorporated herein by reference in its entirety). Reports indicate that RTS, S protects approximately 30% to 50% of children from clinical disease over 18 months. RTS, S has been reported to induce protective antibody and CD4+ T-cell responses, but only negligible CD8+ T cell responses (see, for example, Moris et al. Hum Vaccin Immunother 14:17, 2018, which is incorporated herein by reference in its entirety). Phase III studies of RTS, S delivered as a three-dose series with a booster after 1 yr (year) showed moderate vaccine efficacy in children aged 5 to 17 months preventing 36% of clinical malaria cases over the full study period with a median follow-up of 4 yrs, with a range of 20% in high to 66% in low transmission settings. Furthermore, published literature suggests that protection wanes over time including reports of potential negative efficacy after 5 yrs in children with high malaria exposure (Olotu et al. 2016, N. Engl. J. Med. 374:2519-29, which is incorporated herein by reference in its entirety). Another malaria vaccine is " R21 / Matrix-M." R21 / Matrix-M (R21) is a pre-erythrocytic protein subunit, candidate malaria vaccine based on RTS, S. R21 lacks the excess HBsAg found in RTS, S and comprises only fusion protein moieties (Datoo, Mehreen S et al. Lancet (London, England) vol. 397,10287 (2021): 1809-1818, which is incorporated herein by reference in its entirety). In a Phase 3 clinical trial, R21 was administered as three doses, four weeks apart, with a booster 12 months after the third dose. Trials were performed in two sites, a site with seasonal Malaria transmission and a site with perennial Malaria transmission. Twelve-month vaccine efficacy was approximately 75% at the seasonal sites and 68% at the standard sites for time to first clinical malaria episode (see, for example Datoo, Mehreen S et al., which is incorporated herein by reference in its entirety). RTS, S also initially showed a more favorable vaccine efficacy (e.g., 56%), but this efficacy dropped to approximately 36% after 3 years. Both RTS, S and R21 target the same parasite stage (e.g., pre-erythrocytic), and no vaccine currently exists that targets the asexual stage. Thus, an effective malaria vaccine that targets the asexual stage remains an unmet medical need of critical importance for global health.A. Lifecycle

[0105] During a blood meal, infected mosquitoes inject, along with their anticoagulating saliva, motile parasite forms known as sporozoites into the skin. After deposition in the skin, sporozoites traverse through the skin in search of blood vessels. If they are successful and enter the blood stream, they travel passively with circulating blood until they reach the liver, where they invade and replicate within hepatocytes. This journey to the liver happens very quickly; it can be completed within only a few minutes (Sinnis et al., Parasitol Int. 2007 Sep;56(3): 171-8, which is incorporated herein by reference in its entirety). It is challenging for the host immune system to mount an effective response against the parasite at this stage, as only a small number (thought to be a few hundred at maximum) of sporozoites are injected by the mosquito, with a small fraction of those parasites establishing infection in the liver (Flores-Garcia et al., mBio. 2018 Nov 20;9(6):e02194-18, which is incorporated herein by reference in its entirety).

[0106] When moving from an inoculation site in the skin to the liver, sporozoites traverse host cells (Mota et al., Science 2001 Jan 5;291(5501): 141-4). Sporozoites traverse different types of host cells at the dermis, including fibroblasts and phagocytes (Amino et al., Cell Host Microbe. 2008 Feb 14;3(2):88-96, which is incorporated herein by reference in its entirety), and the liver sinusoidal barrier, containing liver endothelial cells and Kupffer cells (Frevert et al., PLoS Biol 3(6): el92. 2005, which is incorporated herein by reference in its entirety) and sinusoidal endothelial cells (Tavares et al., J Exp Med 2013 May 6;210(5):905-15, which is incorporated herein by reference in its entirety), in order to gain access to hepatocytes. Sporozoites preferentially traverse cells with low-sulfated heparin sulfateproteoglycans (HSPGs) but preferentially invade cells with high-sulfated HSPGs (Coppi et al., Cell Host & Microbe 2, 316-327, November 2007, which is incorporated herein by reference in its entirety).

[0107] Cell traversal was first observed as non-phagocytic entry of P. berghei sporozoites into macrophages followed by "escape" from these cells (Vanderberg et al., J. Euk. Microbiol. 37:528-536, 1990, which is incorporated herein by reference in its entirety). The biochemical, biophysical, and stepwise processes of traversal are still being explored. However, it has been suggested by electron microscopy that host cell rupture occurs upon entry and exit from the host cell (Mota et al., 2001; Tavares et al., 2013, each of which is incorporated herein by reference in its entirety). It has also been shown that P. yoelii sporozoites can enter hepatocytes via a transient vacuole and that host membrane rupture occurs upon cell exit rather than cell entry (Risco-Castillo et al., Cell Host Microbe 2015 Nov ll;18(5):593-603, which is incorporated herein by reference in its entirety).

[0108] Sporozoites also traverse hepatocytes before establishing a productive hepatocyte infection (Mota et al., 2001, which is incorporated herein by reference in its entirety). Several possibilities emerged as to why this occurs. The first hypothesis suggested that migration through hepatocytes primes parasites for invasion by activating apical exocytosis (Mota et al., Nat Med 2002 Nov;8(ll): 1318-22, which is incorporated herein by reference in its entirety). The second theory suggested that traversal releases hepatocyte growth factor (HGF), making neighboring hepatocytes more susceptible to infection (Carrolo et al., Nat Med. 2003 Nov;9(ll):1363-9, which is incorporated herein by reference in its entirety). Lastly, other studies suggest that it takes some time for sporozoites to switch off the machinery for traversal and activate invasion machinery (Amino et al., 2008, Coppi et al., 2007, each of which is incorporated herein by reference in its entirety), and that traversal primarily functions to penetrate cell barriers and avoid phagocytosis en route to the liver (Amino et al., 2008, Coppi et al., 2007, Tavares et al., 2013, each of which is incorporated herein by reference in its entirety).

[0109] Although it has been shown that sporozoites traverse human cells (Behet et al., Malar J 2014 Apr 5;13:136; Cha et al., J Exp Med 2015 Aug 24;212(9): 1391-403; Dumoulin et al., PLoS One 2015 Jun 12;10(6):e0129623; van Schaijk et al., PLoS ONE, 3 (10). e3549 2008; Risco-Castillo et al., 2015, each of which is incorporated herein by reference in its entirety), the molecular basis for the traversal process is largely unstudied.

[0110] Once sporozoites have invaded liver cells, they undergo a phase of massive replication, ultimately differentiating into merozoites, the red blood cell invasive form of the parasite. In approximately one week, a single sporozoite infected hepatocyte can result in the formation of 40,000-60,000 merozoites (Vaughan et al. The Journal of clinical investigation vol. 122(10) 2012 3618-28, which is incorporated herein by reference in its entirety). Merozoites bud from the host hepatocyte in structures called merosomes, released into the bloodstream, which contain up to a thousand merozoites and are hidden from host immune responses due to their host membrane composition. These merosomes ultimately rupture, releasing merozoites into the blood stream, where they invade red blood cells and begin the blood stage of infection, characterized by cyclical ~48 hours asexual replication, rupture and reinvasion, with clinical presentation ~11 days after infectious mosquitos bite. Each mature blood stage parasite can release up to 32 daughter merozoites, which causes parasite density in the blood to increase rapidly.

[0111] P. / axand P. ovale can also enter a dormant state in the liver, the hypnozoite.

[0112] Plasmodium spp. parasites gain entry into red blood cells through specific ligand-receptor interactions mediated by proteins on the surface of the parasite that interact with receptors on the host erythrocyte (mature red blood cell) or reticulocyte (immature red blood cell). While P. falciparum can invade and replicate in erythrocytes and reticulocytes, P. vivax and other species predominantly invade reticulocytes, which are less abundant than erythrocytes. Most of the erythrocyte-binding proteins or reticulocyte-binding proteins that have been associated with invasion exhibit redundancy or high levels of sequence diversity. The PCRCR complex comprised of key proteins Rh5, CyRPA, Ripr, CSS and TRAMP has been identified as essential for parasite invasion with low sequence diversity found in these proteins.

[0113] The invasion of a red blood cell by the merozoite involves interactions of multiple parasite derived proteins (e.g., ligands) with red blood cell (RBC) proteins (e.g., receptors) (Weiss et al., Pios Path., 2015 Feb 27;10.1371, which is incorporated herein by reference in its entirety). The invasion process begins by the merozoite first attaching to theRBC. Merozoite-RBC interaction is further strengthened through merozoite surface protein-1 (MSP1) and unknown RBC proteins, causing some deformation of the RBC surface. After the deformation, interaction of merozoite EBA and Rh protein families (excluding Rh5) with host receptors like CR1 lead to actin-dependent deformation of the RBC membrane and reorientation of the merozoites apical end onto the RBC surface (Geoghehan et al., Nat Commun., 2021 June 15;10.1038 which is incorporated herein by reference in its entirety). This deformation event increases the surface area of interaction between merozoite and RBC membranes, an interaction which can be stabilized by an Rh5 invasion complex comprising Plasmodium thrombospondin-related apical merozoite protein (TRAMP), cysteine-rich small secreted (CSS), cysteine-rich protective antigen (CyRPA), Rh5-interacting protein (Ripr) and Rh5. This Rh5 invasion complex including the five proteins is referred to as the PCRCR complex.

[0114] The creation of the PCRCR complex is thought to begin in the endoplasmic reticulum, where its constituents cysteine-rich small secreted (CSS) protein and Plasmodium thrombospondin-related apical merozoite protein (TRAMP) interact to form a heterodimer referred to as TRAMP-CSS. It is hypothesized that a tetrameric PCRCR complex is formed by TRAMP-CSS trafficking to a secretory organelle called the microneme, where CSS interacts with Rh5-interacting protein (Ripr) and cysteine-rich protective antigen (CyRPA) to form PCRC. At some point after deformation occurs, the merozoite initiates a polarized secretion process that allows the micronemal PCRC complex opportunity to interact with rhoptry protein Rh5, forming the pentameric complex PCRCR. The PCRCR complex, through Rh5, can then bind a host cell receptor, called basigin. PCRCR binding to basigin creates a stable and irreversible platform between the apical end of the merozoite and the deformed RBC surface (Scally et al., Nat Microb., 2022 May 4, which is incorporated herein by reference in its entirety). Once the merozoite-RBC interaction is secured, an open connection between the merozoite's apical tip and the RBC surface is formed which acts as a conduit for Ca2+to flow into the RBC, and for merozoite derived invasion proteins, such as AMA1 and RON2, to help establish the moving junction (Srinivasan et al., Proc Natl Acad Sci., 2011 Jul 25;13275-80, which is incorporated herein by reference in its entirety). It is through this moving junction that the parasite, utilizing its actin-myosin dependent gliding motility, propels itself inside the RBC and establishes a parasitophorous vacuole within which the parasite grows and replicates.

[0115] It has been shown that Rh5 binds basigin with higher affinity when complexed with CyRPA and Ripr to form the RCR complex, and has the highest affinity for basigin when part of the PCRCR complex (Wong et al., Nature., 2019 and Scally et al., Nat Microb., 2022 May 4, each of which are incorporated herein by reference in its entirety).

[0116] Merozoites that egress from red blood cells can invade other red blood cells to continue to the asexual blood stage of the parasite lifecycle. A small percentage of schizonts are already committed to a different fate and merozoites from these schizonts, following invasion of a new red blood cell, will differentiate into the sexual forms of the parasite's life cycle; either male or female, gametocytes. Gametocytes are taken up by the mosquito vector during blood feeding. In the gut of the mosquito, male gametocytes undergo 3 rapid rounds of mitosis to form 8 flagellate microgametes. Female gametocytes mature into macrogametes, egressing from their red blood cell. Male microgametes are motile forms with flagellae and seek the female macrogamete. The male and female gametes fuse, forming a diploid zygote, which matures and differentiates into an ookinete; this motile form secretes chitinases and other lytic proteins such as CelTOS in order to disrupt and traverse through the peritrophic matrix and midgut epithelium to reach the basal lamina where it further differentiates and matures as an oocyst. Oocysts mature over approximately 10 days (depending on the temperature), replicating to form sporozoites that egress the mature oocyst into the hemocoel of the mosquito. Thousands of sporozoites form in a single oocyst and become randomly distributed throughout the hemocoel. These midgut sporozoites have 4 rhoptries, a polarized secretory organelle responsible for releasing factors essential to invasion. It is thought that sporozoites are passively circulated through the mosquito haemolymph until they encounter the salivary glands, where they actively invade the glands. Following invasion of the salivary gland, sporozoites are re-programmed to prepare for liver invasion. It is hypothesized that the salivary gland invasion process utilizes two of the parasite's four rhoptries, reserving the remaining two rhoptries for vertebrate host cell invasion. Evidence of this reprogramming has been demonstrated by the inability of midgut sporozoites (directly from oocysts) to invade hepatocytes, and also by the fact that sporozoites which have successfully invaded a salivary gland are unableto re-invade another salivary gland if presented one. Salivary gland sporozoites alter mosquito behavior and salivary gland function, as less saliva is produced resulting in an increase in mosquito probing behavior, increasing the chances of transmission to a human host via a mosquito bite and continuing the human host's phase of this parasite's life cycle.

[0117] Malaria symptoms typically develop 4-8 days after initial red blood cell invasion. Replication cycle of blood stage parasites within red blood cells lasts approximately 48 hours, ending with hemolysis, and the release of up to 32 daughter merozoites, thereby setting up another round of red blood cell infection. Thus, in synchronous infections (infections that originate from a single infectious bite), fever occurs approximately every 48 hours as red blood cells lyse and release endotoxins en masse, giving P. falciparum the name malignant tertian malaria.

[0118] Some drugs that prevent Plasmodium spp. invasion or proliferation in the liver have prophylactic activity, drugs that block the red blood cell stage are required for the treatment of the symptomatic phase of the disease, and compounds that inhibit the formation of gametocytes or their development in the mosquito (including drugs that kill mosquitoes feeding on blood) are transmission-blocking agents (Phillips, et al. Malaria. Nat Rev Dis Primers 3, 17050 (2017), which is incorporated herein by reference in its entirety).B. Genome

[0119] Since completion of the first sequence of P. falciparum 3D7 genome in 2002, genomic research on malaria parasites has rapidly advanced. Except for a short phase after fertilization in the mosquito midgut, Plasmodium spp. parasites are haploid throughout their life cycle. The genomes of different species range from 20 to 35 megabases, contain 14 chromosomes, a circular plastid genome of approximately 35 kilobases, and multiple copies of a 6 kilobase mitochondrial DNA. Comparison of genomes from different species showed that homologous genes are often found in syntenic blocks arranged in different orders among different chromosomes.

[0120] The adenine-thymine (AT) content of Plasmodium spp. can also be very different, e.g., -80% AT in P. falciparum, P. reichenowi, and P. gallinaceum, -75% AT in rodent malaria parasites; and -60% AT in P. vivax, P. knowiesi, and P. cynomoigi. AT content is often higher in introns and intergenic noncoding regions than in proteincoding exons, with an average of 80.6% AT for the whole P. falciparum genome versus 86.5% for noncoding sequences. The high AT content of P. falciparum reflects large numbers of low-complexity regions, simple sequence repeats, and microsatellites, as well as a highly skewed codon usage bias. Polymorphisms of AT-rich repeats provide abundant markers for linkage mapping of drug resistance genes and for tracing the evolution and structure of parasite populations.

[0121] Malaria parasite genomes carry multigene families that serve important roles in parasite interactions with their hosts, including, for example, antigenic variation, signaling, protein trafficking, and adhesion. Among the gene families, genes encoding P. falciparum erythrocyte membrane protein 1 (PfEMPl) have been studied most extensively. Each individual P. falciparum parasite carries a set of ~60 vars gene in its genome, where switches of gene expression result in antigenic variation and immune evasion. PfEMPl plays an important role in the pathogenesis of clinical developments such as in cerebral and placental malaria, in which it mediates the cytoadherence of infected red blood cells (iRBCs; infected erythrocytes) in the deep tissues. Different PfEMPl molecules bind to various host molecules, including a2-macroglobulin, CD36, chondroitin sulfate A (CSA), complement 1q, CR1, E-selectins and P-selectins, endothelial protein C receptor (EPCR), heparan sulfate, ICAM1, IgM, IgG, PECAM1, thrombospondin (TSP), and VCAM1. Such binding leads to activation of various host inflammatory responses. Hemoglobinopathies, including the hemoglobin C and hemoglobin S trait conditions, interfere with PfEMPl display in knob structures of the iRBCs. This poor display of PfEMPl on the host cell surface offers protection against malaria by reducing the cytoadherence and activation of inflammatory processes that promote the development of severe disease.

[0122] Plasmodium genomes can be highly polymorphic. Early studies demonstrated polymorphisms involving tens to hundreds of kilobases and that the chromosome structure in P. falciparum is largely conserved in central regions but extensively polymorphic is both length and sequence near the telomeres. Much of the subtelomeric variation was explained by recombination within blocks of repetitive sequences and families of genes.C. Plasmodium Proteins1. Rh5 Invasion Complex PolypeptidesRh5

[0123] Rh5 is found in P. falciparum, but not in all other species of Plasmodium that infect humans which contain proteins that perform an analogous function as Rh5. Rh5 orthologues are also found in other species belonging to the Lavarenia subgenus, which includes parasites that infect chimpanzees and gorillas. See, e.g., Ragotte, et al. Trends Parasitol. 36(6) 2020, which is incorporated herein by reference in its entirety. Rh5 is expressed during the mature schizont stages and localizes to the Rhoptry, a polarized secretory organelle of P. falciparum. Rh5 is a protein that is secreted during the invasion process and is believed to be essential for the completion of parasite invasion of the erythrocyte. Rh5 functions downstream of initial parasite attachment to the erythrocyte surface and deformation of the erythrocyte membrane. Upon secretion, Rh5 binds to erythrocyte surface protein basigin, creating an attachment and a stable platform for downstream receptor-ligand interactions and associated invasion events to occur. Binding of Rh5 to basigin is also required for the induction of a spike in calcium within the erythrocyte, which is blocked when merozoites attempt to invade in the presence of anti-Rh5, anti-Ripr, or anti-basigin antibodies or soluble basigin (See, e.g., Ragotte (2020), which is incorporated herein by reference in its entirety).

[0124] Rh5 binds CyRPA, which itself binds Ripr to form an elongated protein trimer, called RCR, on the merozoite surface. Ripr, as part of the RCR, can then bind CSS, which binds TRAMP to form a pentameric complex known as PCRCR. Rh5 has its highest affinity to basigin when part of the PCRCR complex in. Each constituent (i.e., Rh5, CyRPA, Ripr, TRAMP, and CSS) of the PCRCR complex is essential for parasite invasion. See, Scally (2022), which is incorporated herein by reference in its entirety.

[0125] Rh5 is a 63 kDa protein expressed during the mature schizont stage. It is processed and cleaved to a 45 kDa form. The structure of Rh5 reveals a kite-like architecture formed from the coming together of two three-helical bundles. See, e.g., Ragotte (2020), which is incorporated herein by reference in its entirety.

[0126] Rh5 sequences are known (see, e.g., UniProt accession numbers A0A159SK44, A0A159SK99, A0A159SKS8, A0A159SKW8, A0A159SL23, A0A159SL78, A0A159SL96, A0A159SLM7, A0A159SMC8, A0A159SMR9, A0A161FQT0, A0A1B1UZE2, A0A1B1UZE4, A0A1B1UZE5, A0A346RCI1, A0A346RCJ0, A0A346RCJ2, A0A346RCJ3, A0A346RCJ4, A0A346RCK4, A0A346RCK5, A0A346RCK6, A0A346RCK9, B2L3N7, Q8IFM5, each of which is incorporated herein by reference in its entirety), and exemplary Rh5 amino acid sequence is provided in Table 1.Table 1: Exemplary Sequences for Rh5 PolypeptidesSEQ ID Protein Sequence (Amino Acid)NO:1 Rh5 (3D7) MIRIKKKLILTIIYIHLFILNRLSFENAIKKTKNQENNLTLLPIKSTEEEKDDIKNGKDIKKEIDNDKE NIKTNNAKDHSTYIKSYLNTNVNDGLKYLFIPSHNSFIKKYSVFNQINDGMLLNEKNDVKNNEDY KNVDYKNVN FLQYH FKELSNYN IAN SI DI LQEKEG H LDFVII PH YTFLDYYKH LSYN SIYH KSSTY GKCIAVDAFIKKINETYDKVKSKCNDIKNDLIATIKKLEHPYDINNKNDDSYRYDISEEIDDKSEET DDETEEVEDSIQDTDSNHTPSNKKKNDLMNRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICM DMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDMTNILQQSELLLT NLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDM SN EYSLFITSDH LRQM LYNTFYSKEKH LN NI FH H LIYVLQM KFN DVPI KM EYFQTYKKN KPLTQCysteine-Rich Protective Antigen (CyRPA)

[0127] Cysteine-Rich Protective Antigen (CyRPA) is a 43 kDa protein with a predicted N-terminal secretion signal. CyRPA is a critical component of the pentameric PCRCR complex and is positioned between Ripr and RH5 in the complex. CyRPA mediated Rh5 binding to basigin is important for stabilizing the parasite-erythrocyte membrane interaction which allows downstream invasion events, like Ca2+increase in the erythrocyte cytosol and establishment of tight / moving junctions, to occur. CyRPA is highly conserved, with only a single SNP above 5% prevalence in thegeneral population. CyRPA has poor sero-reactivity from natural exposure (See, e.g., Ragotte (2020) and Cowman (2018) each of which is incorporated herein by reference in its entirety).Rh5-interacting Protein (Ripr)

[0128] Rh5-interacting Protein (Ripr) is an approximately 120 kDa protein and localized to micronemes during the schizont stage of the Plasmodium life cycle. The full-length 120 kDa protein is processed into two fragments of similar size, an N-terminal fragment (including EGF domains 1 and 2) and a C-terminal fragment (including EGF domains 3-10). Ripr colocalizes with Rh5 and CyRPA during parasite invasion at the junction between merozoites and erythrocyte. Parasites with conditional knockouts of ΔRipr induce membrane deformation, but cannot complete invasion (See, e.g., Ragotte (2020) which is incorporated herein by reference in its entirety).Cysteine-Rich Small Secreted Protein (CSS)

[0129] Cysteine-rich small secreted protein (CSS) is a 290 amino acid cryptic 6-Cys protein comprised of two degenerate 6-Cys domains. Proteins belonging to this family commonly mediate extracellular protein-protein interactions, a role that has been confirmed to occur in P. falciparum. CSS is capable of biding both TRAMP and Ripr. CSS interaction with TRAMP occurs in the endoplasmic reticulum, whereas CSS interaction with Ripr occurs after trafficking to the micronemes. CSS Interaction with TRAMP and Ripr are involved in invasion and the formation of the PCRC complex, which complexes with Rh5, completing the pentameric complex and facilitating the interaction of Rh5 with basigin. Similar to TRAMP, nanobodies against CSS have been shown to inhibit invasion, indicating its role in the invasion process as an essential component of the PCRCR complex. Nanobodies which inhibit CSS binding to TRAMP or Ripr do not inhibit invasion, confirming the interaction of these proteins before exposure to the merozoite surface. The inhibitory activity of nanobodies against CSS are thought to occur by potentially blocking Ripr and / or Rh5 insertion into the RBC membrane and / or disrupting the conformational changes of the PCRCR complex required for invasion, although the exact mechanism of inhibition is not entirely known. Contrary to this observation and of immunological significance, polyclonal antibodies to CSS and TRAMP do not exhibit any neutralizing activity, indicating the immunodominance of non-neutralizing epitopes of CSS and TRAMP that bias antibody selection (in a natural setting) away from epitopes with neutralizing capability. This observation provides the molecular rationale for identifying invasion inhibitory epitopes on CSS and TRAMP which are highly conserved in P. falciparum, for usage in vaccine platforms. See Cowman (2017), which is incorporated herein by reference in its entirety.Plasmodium Thrombospondin-Related Apical Merozoite Protein (TRAMP)

[0130] Plasmodium thrombospondin-related apical merozoite protein (TRAMP) is a 352 amino acid protein that localizes to developing micronemes and relocates to the merozoite surface during invasion and is understood to be important for host cell invasion by P. falciparum. Orthologs to the gene encoding TRAMP is present in all malaria parasite species examined, indicating a conserved role in host invasion and making it a potential target for therapeutic intervention. TRAMP interacts with cysteine-rich small secreted (CSS) protein to form a heterodimer known as TRAMPCSS which in turn can, through CSS, interact with Ripr and CyRPA to form a tetrameric complex called PCRC, that ultimately interacts with Rh5 forming a pentameric complex PCRCR which presents RH5 on the surface of merozoites and enhances Rh5 binding to host basigin, an essential step for P. falciparum RBC invasion. Nanobodies against TRAMP significantly reduce merozoite invasion providing evidence for the potential for TRAMP as a blood stage vaccine candidate (Scally et al., Nat Microb., 2022 May 4, which is incorporated herein by reference in its entirety).P113

[0131] P113 is a glycosylphosphatidylinositol (GPI)-linked protein that interacts directly with the N terminus of unprocessed Rh5, which has been thought to provide a mechanism by which the Rh5 invasion complex is tethered to the merozoite surface. However, recent findings have shown that P113 may not be essential for P. falciparum growth, as other proteins and complexes exist that can perform the Rh5 membrane tethering function of P113. See, e.g., Ragotte (2020) and Scally (2022). In addition to its canonical role, P113 is present in the parasitophorous vacuole (PV) and has been shown to interact with parasite export complexes like PTEX and EPIC, which are important for the transport of parasite effector proteins across the parasitophorous vacuolar membrane (PVM). P113 has also beenshown to play a role in PVM architecture, as the deletion of this gene leads to a disruption in PVM morphology. P113 orthologues are found in all Plasmodium species sequenced thus far, which is suggestive of common and conserved functions (Bullen et al. (2022) Molecular Microbiology 117:1245-1262, which is incorporated herein by reference in its entirety). Despite this fact, in P. berghei, P113 knockout parasites were viable, indicating the protein was not essential for asexual blood stage growth and invasion. The knockout parasites do, however, display defects in natural sporozoite transmission, leading to delayed patency in infected mice (Offeddu et al. (2014) Mol. Biochem. Parasitology 193:101-109, which is incorporated herein by reference in its entirety).II. Plasmodium polypeptide constructs

[0132] The present disclosure, among other things, utilizes RNA technologies as a modality to express one or more Plasmodium polypeptide construct that includes one or more malarial proteins, or one or more portions thereof, described herein. For example, in some embodiments, a Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 polypeptides or portions thereof (e.g., antigenic portions thereof). In some embodiments, a portion of a Rh5 polypeptide can be a characteristic portion of a Rh5 polypeptide. In some embodiments, a Plasmodium polypeptide construct additionally includes one or more additional amino acid sequences, such as a secretory signal (e.g., a heterologous secretory signal), a transmembrane region (e.g., a heterologous transmembrane region), a multimerization region, and / or a linker, as described herein.A. Rh5 Polypeptides or Portions Thereof

[0133] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more Plasmodium Rh5 polypeptides or portions thereof (e.g., antigenic portions thereof).

[0134] A full-length wild-type Rh5 comprises, in N-terminal to C-terminal order, a signal peptide (corresponding to amino acids 1-25), an N-terminal disordered domain (corresponding to amino acids 26-139), an N-terminal ordered domain (corresponding to amino acids 140-247), a linking disordered domain (corresponding to amino acids 248-296), and a C-terminal ordered domain (corresponding to amino acids 297-526).

[0135] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more polypeptides or antigenic portions of an Rh5 polypeptide, e.g., Plasmodium Rh5 polypeptide, e.g., P. falciparum Rh5 (SEQ ID NO: 1), or a variant thereof (e.g., a glycosylation variant). In some embodiments, an Rh5 polypeptide may include an N-terminal disordered domain, an N-terminal ordered domain, a linking disordered domain, a C-terminal ordered domain, or a combination thereof. A portion of Rh5 (or Rh5 polypeptide portion) (e.g., an antigenic portion of Rh5 or Rh5 antigenic portion) may refer to parts of an Rh5 polypeptide domain or parts spanning two or more Rh5 polypeptide domains.

[0136] In some embodiments, a Plasmodium polypeptide construct comprises one or more antigenic portions of Rh5. In some embodiments, one or more antigenic portions of Rh5 comprise one or more N-terminal ordered domains. In some embodiments, one or more antigenic portions of Rh5 comprise one or more C-terminal ordered domains. In some embodiments, a Plasmodium Rh5 polypeptide or antigenic portion thereof comprises at least 25, 30, 35, 40, or 45 contiguous amino acids of an Rh5 ordered domain (e.g., an N-terminal ordered domain or a C-terminal ordered domain), or a variant thereof (e.g., a glycosylation variant).

[0137] In some embodiments, a Plasmodium Rh5 antigenic portion comprises an N-terminal ordered domain that corresponds to amino acids 145-240, amino acids 140-247, amino acids 145-247, amino acids 155-240, and amino acids 155-247 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof (e.g., glycosylation variants). In some embodiments, a Plasmodium Rh5 antigenic portion comprises a C-terminal ordered domain that corresponds to amino acids 297-526, amino acids 297-506, amino acids 297-497 of a wild-type Rh5 sequence (SEQ ID NO: 1), or a variant thereof (e.g., a glycosylation variant). In some embodiments, one or more antigenic portions of Rh5 comprise (i) an N-terminal ordered domain that corresponds to amino acids 145-247 or amino acids 155-247 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof and / or (ii) a C-terminal ordered domain that corresponds to amino acids 297-506 or amino acids 297-497 of a wild-type Rh5 sequence (SEQ ID NO: 1), or a variant thereof.

[0138] 145-240 N-Terminal Ordered Domain of Rh5 (SEQ ID NO: 22) LQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKINETYDKVKSKCNDIKNDLIATI KKLEH

[0139] 140-247 N-Terminal Ordered Domain of Rh5 (SEQ ID NO: 31) KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKINETYDKVKSKCNDIKN DLIATI KKLEH

[0140] 145-247 N-Terminal Ordered Domain of Rh5 (SEQ ID NO: 20) LQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKINEAYDKVKSKCNDIKNDLIATI KKLEHPYDINNK

[0141] 155-247 N-Terminal Ordered Domain of Rh5 (SEQ ID NO: 26) YNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKINETYDKVKSKCNDIKNDLIATI KKLEH

[0142] 155-240 N-Terminal Ordered Domain of Rh5 (SEQ ID NO: 24) YNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKINETYDKVKSKCNDIKNDLIATI KKLEH

[0143] 297-506 C-Terminal Ordered Domain of Rh5 (SEQ ID NO: 45) NRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFN

[0144] 297-497 C-Terminal Ordered Domain of Rh5 (SEQ ID NO: 36) NRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHL

[0145] 297-526 C-Terminal Ordered Domain of Rh5 (SEQ ID NO: 39) NRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0146] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise one or more unpaired cysteines. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise two unpaired cysteines. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a cysteine at position 203, as numbered according to SEQ ID NO: 1. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a tyrosine at position 203, as numbered according to SEQ ID NO: 1. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise a tyrosine at position 203, as numbered according to SEQ ID NO: 1.

[0147] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise one, two, three or more N-linked glycosylation sites. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise three N-linked glycosylation sites. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise two N-linked glycosylation sites. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation. In some embodiments, an amino acid substitution that prevents glycosylation comprises a NX[T / S] to QX[T / S] substitution. In some embodiments, an amino acid substitution that prevents glycosylation comprises a NX[T / S] to NXA substitution. In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise an amino acid substitution at all of the N-linked glycosylation sites, wherein the amino acid substitution prevents glycosylation.

[0148] In some embodiments, a Plasmodium Rh5 antigenic portion comprises a N-terminal ordered domain that corresponds to amino acids 140-247 of a wild-type Rh5 (SEQ ID NO: 1) (or amino acids 140-247 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions). In some embodiments, an N-terminal ordered domain corresponds to amino acids 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a cysteine at position 203. In some embodiments, an N-terminal ordered domain corresponds to amino acids 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203. In some embodiments, an N-terminal ordered domain corresponds to aminoacids 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N linked glycosylation site at N214. In some embodiments, an N-terminal ordered domain corresponds to amino acids 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 214. In some embodiments, an amino acid substitution that prevents glycosylation at position 214 comprises a NX[T / S] to QX[T / S] substitution. In some embodiments, an amino acid substitution prevents glycosylation at position 214 comprises a NX[T / S] to NXA substitution. In some embodiments, an amino acid substitution at position 216 prevents glycosylation at position 214. In some embodiments, an amino acid substitution that prevents glycosylation at position 214 comprises T216A.

[0149] In some embodiments, an N-terminal ordered domain corresponds to amino acids 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an alanine at position 216.

[0150] 140-247 N-Terminal Ordered Domain of Rh5 - T216A (SEQ ID NO: 30) KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKINEAYDKVKSKCNDIKN DLIATIKKLEHPYDINNK

[0151] In some embodiments, an N-terminal ordered domain corresponds to amino acids 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and glutamine at position 214.

[0152] 140-247 N-terminal Ordered Domain of Rh5 - C203Y - N214Q (SEQ ID NO: 32) KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKIQETYDKVKSKCNDIKN DLIATIKKLEHPYDINNK

[0153] In some embodiments, an N-terminal ordered domain corresponds to amino acids 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203, a glutamine at position 214, and a threonine at position 231.

[0154] 140-247 N-terminal Ordered Domain of Rh5 - C203Y - N214Q - L231T (SEQ ID NO: 33) KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKIQETYDKVKSKCNDIKN DTIATIKKLEHPYDINNK

[0155] In some embodiments, an N-terminal ordered domain corresponds to amino acids 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203, a glutamine at position 214, and a threonine at position 231, an asparagine at position 226, a threonine at position 228, an asparagine at position 237, a leucine at position 238, and a threonine at position 239.

[0156] 140-247 N-terminal Ordered Domain of Rh5 - C203Y - N214Q - L231T - D226N - K228T - K237N - L238L - E239T (SEQ ID NO: 34) KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKIQETYDKVKSKCNNITN DLIATIKNLTHPYDINNK

[0157] In some embodiments, a Plasmodium Rh5 antigenic portion comprises a N-terminal ordered domain that corresponds to amino acids 145-247 of a wild-type Rh5 (SEQ ID NO: 1) (or amino acids 145-247 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions). In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a cysteine at position 203. In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203. In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N linked glycosylation site at N214. In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 214. In some embodiments, an amino acid substitution that prevents glycosylation at position 214 comprises a NX[T / S] to QX[T / S] substitution. In some embodiments, an amino acid substitution prevents glycosylation at position 216 comprises a NX[T / S] to NXA substitution. In some embodiments, an amino acid substitution at position 216 prevents glycosylation at position 214. In some embodiments, an amino acid substitution that prevents glycosylation at position 214 comprises T216A.

[0158] In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and glutamine at position 214.145-247 N-Terminal Ordered Domain of Rh5- C203Y- N214Q (SEQ ID NO: 19) LQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKIQETYDKVKSKCNDIKNDLIATI KKLEHPYDINNK

[0159] In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an alanine at position 216.

[0160] 145-247 N-Terminal Ordered Domain of Rh5 - T216A (SEQ ID NO: 28) LQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKINEAYDKVKSKCNDIKNDLIATI KKLEHPYDINNK

[0161] In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and alanine at position 216.

[0162] 145-247 N-Terminal Ordered Domain of Rh5 - C203Y - T216A (SEQ ID NO: 29) LQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKINEAYDKVKSKCNDIKNDLIATI KKLEHPYDINNK

[0163] In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a leucine at position 157, a glutamate at position 183, an alanine at position 216, and a lysine at position 233.

[0164] 145-247 N-Terminal Ordered Domain of Rh5 - I157L - D183E - T216A - A233K SEQ ID NO: 27) LQYHFKELSNYNLANSIDILQEKEGHLDFVIIPHYTFLEYYKHLSYNSIYHKSSTYGKCIAVDAFIKKINEAYDKVKSKCNDIKNDLIKTI KKLEHPYDINNK

[0165] In some embodiments, a Plasmodium Rh5 antigenic portion comprises a N-terminal ordered domain that corresponds to amino acids 145-240 of a wild-type Rh5 (SEQ ID NO: 1) (or amino acids 145-240 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions). In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-240 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a cysteine at position 203. In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-240 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203. In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-240 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N linked glycosylation site at N214. In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-240 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 214. In some embodiments, an amino acid substitution that prevents glycosylation at position 214 comprises a NX[T / S] to QX[T / S] substitution.

[0166] In some embodiments, an N-terminal ordered domain corresponds to amino acids 145-240 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and glutamine at position 214.145-240 N-Terminal Ordered Domain of Rh5 - C203Y- N214Q (SEQ ID NO: 21) LQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKIQETYDKVKSKCNDIKNDLIATI KKLEH

[0167] In some embodiments, a Plasmodium Rh5 antigenic portion comprises an N-terminal ordered domain that corresponds to amino acids 155-247 of a wild-type Rh5 (SEQ ID NO: 1) (or amino acids 155-247 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions). In some embodiments, an N-terminal ordered domain corresponds to amino acids 155-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a cysteine at position 203. In some embodiments, an N-terminal ordered domain corresponds to amino acids 155-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203. In some embodiments, an N-terminal ordered domain corresponds to amino acids 155-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N-linked glycosylation site at N214. In some embodiments, an N-terminal ordered domain corresponds to amino acids 155-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 214. In some embodiments, an amino acid substitution that prevents glycosylation at position 214 comprises a NX[T / S] to QX[T / S] substitution. In someembodiments, an N-terminal ordered domain corresponds to amino acids 155-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and a glutamine at position 214.

[0168] 155-247 N-terminal Ordered Domain of Rh5 - C203Y - N214Q (SEQ ID NO: 25) YNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKIQETYDKVKSKCNDIKNDLIATIKKLEHPYDIN NK

[0169] In some embodiments, a Plasmodium Rh5 antigenic portion comprises an N-terminal ordered domain that corresponds to amino acids 155-240 of a wild-type Rh5 (SEQ ID NO: 1) (or amino acids 155-240 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions). In some embodiments, an N-terminal ordered domain corresponds to amino acids 155-240 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a cysteine at position 203. In some embodiments, an N-terminal ordered domain corresponds to amino acids 155-240 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203. In some embodiments, an N-terminal ordered domain corresponds to amino acids 155-240 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N-linked glycosylation site at N214. In some embodiments, an N-terminal ordered domain corresponds to amino acids 155-240 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 214. In some embodiments, an amino acid substitution that prevents glycosylation at position 214 comprises a NX[T / S] to QX[T / S] substitution.

[0170] In some embodiments, an N-terminal ordered domain corresponds to amino acids 155-240 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203 and a glutamine at position 214.

[0171] 155-240 N-terminal Ordered Domain of Rh5 - C203Y - N214Q (SEQ ID NO: 23) YNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKIQETYDKVKSKCNDIKNDLIATIKKLEH

[0172] In some embodiments, a Plasmodium Rh5 antigenic portion comprises a C-terminal ordered domain that corresponds to amino acids 297-506 of a wild-type Rh5 (SEQ ID NO: 1) (or amino acids 297-506 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions).

[0173] In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-506 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N linked glycosylation site at T299. In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-506 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 297. In some embodiments, an amino acid substitution that prevents glycosylation at position 297 comprises a NX[T / S] to QX[T / S] substitution. In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-506 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 299. In some embodiments, an amino acid substitution prevents glycosylation at position 299 comprises a NX[T / S] to NXA substitution.

[0174] In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-506 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a glutamine at position 297. In some embodiments, said C-terminal domain comprises a lysine at position 458.

[0175] 297-506 C-terminal Ordered Domain of Rh5 - N297Q (SEQ ID NO: 37) QRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFN

[0176] 297-506 C-terminal Ordered Domain of Rh5 - N297Q - R458K (SEQ ID NO: 150) QRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKKILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFN

[0177] In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-506 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an alanine at position 299. In some embodiments, said C-terminal domain comprises a lysine at position 458.

[0178] 297-506 C-terminal Ordered Domain of Rh5 - T299A (SEQ ID NO: 38)NRAFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFN

[0179] 297-506 C-terminal Ordered Domain of Rh5 - T299A - R458K (SEQ ID NO: 155) NRAFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKKILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFN

[0180] In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-506 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an alanine at position 299, an aspartate at position 304, an asparagine at position 312, a phenylalanine at position 314, a asparagine at position 316, an asparagine at position 330, an alanine at position 370, an asparagine at position 381, a lysine at position 384, a lysine at position 392, an asparagine at position 395, an glutamic acid at position 398, a lysine at position 458, a lysine at position 464, an alanine at position 467, and a leucine at position 505.

[0181] 297-506 C-terminal Ordered Domain of Rh5 - T299A - M304F - K312N - L314F - K316N - M330N - S370A - S381N - T384K - L392K - T395N - N398E - R458K - N464K - S467A - F505L (SEQ ID NO: 44) NRAFKKMFDEYNTKKNKFINCIKNHENDFNKICNDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILAVKSKNLNKDLNDM KNILQQSEKLLNNLEKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKKILDMSKEYALFITSDH LRQ M LYNTFYS KEKH LN N I FH H LI YVLQM KLN

[0182] In some embodiments, a Plasmodium Rh5 antigenic portion comprises a C-terminal ordered domain that corresponds to amino acids 297-526 of a wild-type Rh5 (SEQ ID NO: 1) (or amino acids 297-526 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions).

[0183] In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N linked glycosylation site at N297. In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 297. In some embodiments, an amino acid substitution that prevents glycosylation at position 297 comprises a NX[T / S] to QX[T / S] substitution. In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-506 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 299. In some embodiments, an amino acid substitution prevents glycosylation at position 299 comprises a NX[T / S] to NXA substitution.

[0184] In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an alanine at position 299. In some embodiments, said C-terminal domain comprises a lysine at position 458.

[0185] 297-526 C-terminal Ordered Domain of Rh5 - T299A (SEQ ID NO: 40) NRAFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0186] 297-526 C-terminal Ordered Domain of Rh5 - T299A - R458K (SEQ ID NO: 148) NRAFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKKILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0187] In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a glutamine at position 297. In some embodiments, said C-terminal domain comprises a lysine at position 458.

[0188] 297-526 Ordered Domain of Rh5 - N297Q (SEQ ID NO: 41)QRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0189] 297-526 Ordered Domain of Rh5 - N297Q (SEQ ID NO: 149) QRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKKILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0190] In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a glutamine at position 297, a threonine at position 297, and a threonine at position 398. In some embodiments, said C-terminal domain comprises a lysine at position 458.

[0191] 297-526 Ordered Domain of Rh5 - N297Q - L387T - N398T (SEQ ID NO: 42) QRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNITQQSELLLTNLTKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0192] 297-526 Ordered Domain of Rh5 - N297Q - L387T - N398T - R458K (SEQ ID NO: 153) QRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNITQQSELLLTNLTKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKKILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0193] In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a glutamine at position 297, an asparagine at position 302, a leucine at position 303, a threonine at position 304, a threonine at position 387, a threonine at position 398, an asparagine at position 495, a leucine at position 496, and a threonine at position 497. In some embodiments, said C-terminal domain comprises a lysine at position 458.

[0194] 297-526 Ordered Domain of Rh5 - N297Q - K302N - M303L - M304T - L387T - N398T - H495N - H496L - L497T (SEQ ID NO: 43) QRTFKNLTDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNITQQSELLLTNLTKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFNLTIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0195] 297-526 Ordered Domain of Rh5 - N297Q - K302N - M303L - M304T - L387T - N398T - H495N - H496L - L497T - R458K (SEQ ID NO: 154) QRTFKNLTDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNITQQSELLLTNLTKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKKILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFNLTIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0196] In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-497 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N linked glycosylation site at N297. In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-497 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 297. In some embodiments, an amino acid substitution that prevents glycosylation at position 297 comprises a NX[T / S] to QX[T / S] substitution.

[0197] In some embodiments, a C-terminal ordered domain corresponds to amino acids 297-497 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a glutamine at position 297.

[0198] 297-497 C-terminal Ordered Domain of Rh5 - N297Q (SEQ ID NO: 151) QRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDM TNILQQSELLLTNLNKKMGSYIYIDTIKFIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDH LRQMLYNTFYSKEKHLNNIFHHL

[0199] In some embodiments, one or more antigenic portions of Rh5 comprise one or more linking disordered domains. In some embodiments, one or more antigenic portions of Rh5 comprise one or more N-terminal disordereddomains. In some embodiments, a Plasmodium Rh5 polypeptide or antigenic portion thereof comprises at least 25, 30, 35, 40, or 45 contiguous amino acids of an Rh5 disordered domain (e.g., an N-terminal disordered domain or a linking disordered domain), or a variant thereof (e.g., a glycosylation variant).

[0200] In some embodiments, a Plasmodium Rh5 antigenic portion comprises a linking disordered domain that corresponds to amino acids 248-296 of a wild-type Rh5 (SEQ ID NO: 1), or a variant thereof (e.g., glycosylation variants). In some embodiments, a Plasmodium Rh5 antigenic portion comprises an N-terminal disordered domain that corresponds to amino acids 26-139 of a wild-type Rh5 sequence (SEQ ID NO: 1), or a variant thereof (e.g., a glycosylation variant). In some embodiments, one or more antigenic portions of Rh5 comprise (i) a linking disordered domain that corresponds to amino acids 248-296 of a wild-type Rh5 sequence (SEQ ID NO: 1), or a variant thereof; and / or (ii) an N-terminal disordered domain that corresponds to amino acids 26-139 of a wild-type Rh5 sequence (SEQ ID NO:1), or a variant thereof.

[0201] In some embodiments, a linking disordered domain corresponds to amino acids 248-296 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N linked glycosylation site at N284. In some embodiments, a linking disordered domain corresponds to amino acids 248-296 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 284. In some embodiments, an amino acid substitution that prevents glycosylation at position 284 comprises a NX[T / S] to QX[T / S] substitution. In some embodiments, a linking disordered domain corresponds to amino acids 248-296 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 286. In some embodiments, an amino acid substitution prevents glycosylation at position 286 comprises a NX[T / S] to NXA substitution.

[0202] 248-296 Linking Disordered Domain (SEQ ID NO: 49) NDDSYRYDISEEIDDKSEETDDETEEVEDSIQDTDSNHTPSNKKKNDLM

[0203] In some embodiments, a linking disordered domain corresponds to amino acids 248-296 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a glutamine at position 284.

[0204] 248-296 Linking Disordered Domain- N284Q (SEQ ID NO: 46) NDDSYRYDISEEIDDKSEETDDETEEVEDSIQDTDSQHTPSNKKKNDLM

[0205] In some embodiments, a linking disordered domain corresponds to amino acids 248-296 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a alanine at position 286.

[0206] 248-296 Linking Disordered Domain of Rh5 - T286A (SEQ ID NO: 48) NDDSYRYDISEEIDDKSEETDDETEEVEDSIQDTDSNHAPSNKKKNDLM

[0207] In some embodiments, an N-terminal disordered domain corresponds to amino acids 26-139 of a wildtype Rh5 (SEQ ID NO: 1). In some embodiments, an N-terminal disordered domain corresponds to amino acids 26-139 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an N linked glycosylation site at N38. In some embodiments, an N-terminal disordered domain corresponds to amino acids 26-139 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 38. In some embodiments, an amino acid substitution that prevents glycosylation at position 38 comprises a NX[T / S] to QX[T / S] substitution. In some embodiments, an N-terminal disordered domain corresponds to amino acids 26-139 of a wild-type Rh5 (SEQ ID NO: 1) and comprises an amino acid substitution that prevents glycosylation at position 40. In some embodiments, an amino acid substitution prevents glycosylation at position 40 comprises a NX[T / S] to NXA substitution.

[0208] 26-139 N-Terminal Disordered Domain of Rh5 (SEQ ID NO: 52) ENAIKKTKNQENNLTLLPIKSTEEEKDDIKNGKDIKKEIDNDKENIKTNNAKDHSTYIKSYLNTNVNDGLKYLFIPSHNSFIKKYSVFN QINDGMLLNEKNDVKNNEDYKNVDY

[0209] In some embodiments, an N-terminal disordered domain corresponds to amino acids 26-139 of a wildtype Rh5 (SEQ ID NO: 1) and comprises a glutamine at position 38.

[0210] 26-139 N-Terminal Disordered Domain of Rh5 - N38Q (SEQ ID NO: 51) ENAIKKTKNQENQLTLLPIKSTEEEKDDIKNGKDIKKEIDNDKENIKTNNAKDHSTYIKSYLNTNVNDGLKYLFIPSHNSFIKKYSVFN QINDGMLLNEKNDVKNNEDYKNVDY

[0211] In some embodiments, an N-terminal disordered domain corresponds to amino acids 26-139 of a wildtype Rh5 (SEQ ID NO: 1) and comprises an alanine at position 40.

[0212] 26-139 N-Terminal Disordered Domain of Rh5 - T40A (SEQ ID NO: 50) ENAIKKTKNQENNLALLPIKSTEEEKDDIKNGKDIKKEIDNDKENIKTNNAKDHSTYIKSYLNTNVNDGLKYLFIPSHNSFIKKYSVFN QINDGMLLNEKNDVKNNEDYKNVDY

[0213] In some embodiments, one or more Plasmodium Rh5 antigenic portions comprise two Rh5 ordered domains (e.g., an N-terminal ordered domain or a C-terminal ordered domain) and one Rh5 disordered domain (e.g., an N-terminal disordered domain or a linking disordered domain).

[0214] In some embodiments, a Plasmodium polypeptide construct does not include an Rh5 disordered domain (e.g., an N-terminal disordered domain or a linking disordered domain). In some embodiments, a Plasmodium Rh5 polypeptide or antigenic portion thereof does not comprise an N-terminal disordered domain that corresponds to amino acids 26-139 of a wild-type Rh5 (SEQ ID NO: 1) and / or a linking disordered domain that corresponds to amino acids 248-296 of a wild-type Rh5 (SEQ ID NO: 1). In some embodiments, a Plasmodium polypeptide construct comprises one or more Plasmodium Rh5 polypeptides or antigenic portions thereof comprising an N-terminal ordered domain and a C-terminal ordered domain. In some embodiments, a Plasmodium polypeptide construct comprises two Plasmodium Rh5 antigenic portions, where one antigenic portion comprises an N-terminal ordered domain and one antigenic portion comprises a C-terminal ordered domain. In some embodiments, a Plasmodium polypeptide construct comprises: (i) an N-terminal ordered domain that corresponds to amino acids 155-240, 155-247, 145-240, 145-247, or 140-247 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof and (ii) a C-terminal ordered domain that corresponds to amino acids 297-497, 297-506, or 297-526 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof. In some embodiments, a Plasmodium polypeptide construct comprises an N-terminal ordered domain and a C-terminal ordered domain that are directly adjacent to one another.

[0215] In some embodiments, a Plasmodium polypeptide construct comprises an Rh5 polypeptide comprising exactly two antigenic portions of Rh5, where the two antigenic portions comprise or consist of an N-terminal domain and a C-terminal domain. In some embodiments, the N-terminal domain corresponds to amino acids 155-240, 155-247, 145-240, 145-247, or 140-247 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof, and the C-terminal domain corresponds to amino acids 297-497, 297-506, or 297-526 of a wild-type Rh5 (SEQ ID NO: 1) or variants thereof.

[0216] In some embodiments, a Plasmodium polypeptide construct comprising an N-terminal ordered domain and a C-terminal ordered domain, where the N-terminal ordered domain corresponds to amino acids 155-240, 155-247, 145-240, 145-247, or 140-247 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a tyrosine at position 203, a glutamine at position 214, and / or an alanine at position 216, and where a C-terminal ordered domain corresponds to amino acids 297-497, 297-506, or 297-526 of a wild-type Rh5 (SEQ ID NO: 1) and comprises a glutamine at position 297.

[0217] In some embodiments, a Plasmodium polypeptide construct does not include a linking disordered domain or variant thereof. In some embodiments, a Plasmodium polypeptide construct does not include an N-terminal disordered domain or variant thereof.B. Secretory Signals

[0218] In some embodiments, a Plasmodium polypeptide construct described herein includes a secretory signal, e.g., that is functional in mammalian cells. In some embodiments, a utilized secretory signal is a heterologous secretory signal. In some embodiments, a heterologous secretory signal comprises or consists of a non-human secretory signal. In some embodiments, a heterologous secretory signal comprises or consists of a viral secretory signal. In some embodiments, a viral secretory signal comprises or consists of an HSV secretory signal (e.g., an HSV-1 or HSV-2 secretory signal). In some embodiments, an HSV secretory signal comprises or consists of an HSV glycoprotein D (gD) secretory signal.

[0219] In some embodiments, a secretory signal comprises or consists of an Ebola virus secretory signal. In some embodiments, an Ebola virus secretory signal comprises or consists of an Ebola virus spike glycoprotein (SGP) secretory signal.

[0220] In some embodiments, a secretory signal is characterized by a length of about 15 to 30 amino acids.

[0221] In many embodiments, a secretory signal is positioned at the N-terminus of a Plasmodium polypeptide construct described herein. In some embodiments, a secretory signal preferably allows secretion of a Plasmodium polypeptide construct from the cell or transport of a Plasmodium polypeptide construct with which it is associated into a defined cellular compartment, preferably a cell surface, endoplasmic reticulum (ER) or endosomal-lysosomal compartment.

[0222] In some embodiments, a secretory signal comprises or consists of a Plasmodium secretory signal. In some embodiments, a Plasmodium secretory signal comprises or consists of a Plasmodium CSP secretory signal (e.g., from Plasmodium falciparum, e.g., from Plasmodium falciparum isolate 3D7 (SEQ ID NO: 57)). In some embodiments, a Plasmodium secretory signal comprises a Plasmodium Rh5 secretory signal.

[0223] In some embodiments, a secretory signal is selected from an S1S2 secretory signal (aa 1-19), an immunoglobulin secretory signal (aa 1-22), a human SPARC secretory signal, a human insulin isoform 1 secretory signal, a human albumin secretory signal, etc. Those skilled in the art will be aware of other secretory signal such as, for example, as disclosed in W02017 / 081082, which is incorporated herein by reference in its entirety (e.g., SEQ ID NOs: 1-1115 and 1728, or fragments or variants thereof). In some embodiments, a Plasmodium polypeptide construct described herein does not comprise a secretory signal.

[0224] In some embodiments, a secretory signal is one listed in Table 2, or a secretory signal having 1, 2, 3, 4, or 5 amino acid differences relative thereto. In some embodiments, a signal sequence is selected from those included in the Table 2 below and / or those encoded by the sequences in Table 3 below.Table 2: Exemplary secretory signalsSEQ ID Signal Sequence (Amino Acid)NO:54 HSV-1 gD SP MGGAAARLGAVILFVVIVGLHGVRSKY55 HSV-2 gD SP MGRLTSGVGTAALLVVAVGLRVVCA56 HSV-2 MG RLTSGVGTAALLVVAVG LRVVCAKYA57 Csp (isolate 3D7) MMRKLAILSVSSFLFVEA58 HSV-1 gD SP 3 MGGAAARLGAVILFVVIVGLHGVRG59 Ebola spike glycoprotein GP MGVTGILQLPRDRFKRTSFFLWVIILFQRTFS60 SARS-CoV-2-S MFVFLVLLPLVSSQCVNLT61 human Ig heavy chain signal peptide MDWIWRILFLVGAATGAHSQM(huSec)62 HuIgGk signal peptide METPAQLLFLLLLWLPDTTG63 IgE heavy chain epsilon-1 signal peptide MDWTWILFLVAAATRVHS64 Japanese encephalitis PRM signal MLGSNSGQRVVFTILLLLVAPAYSsequence65 VSVg protein signal sequence MKCLLYLAFLFIGVNCA66 TRIO MCRGLSAVLILLVSLSAQLHVVVG67 human Ig heavy chain signal peptide 1 MELGLSWIFLLAILKGVQC68 human Ig heavy chain signal peptide 2 MELGLRWVFLVAILEGVQC69 human Ig heavy chain signal peptide 3 MKHLWFFLLLVAAPRWVLS70 human Ig heavy chain signal peptide 4 MDWTWRILFLVAAATGAHSSEQ ID Signal Sequence (Amino Acid)NO:71 human Ig heavy chain signal peptide 5 MDWTWRFLFVVAAATGVQS72 human Ig heavy chain signal peptide 6 MEFGLSWLFLVAILKGVQC73 human Ig heavy chain signal peptide 7 MEFGLSWVFLVALFRGVQC74 human Ig heavy chain signal peptide 8 MDLLHKNMKHLWFFLLLVAAPRWVLS75 human Ig kappa chain signal peptide 1 MDMRVPAQLLGLLLLWLSGARC76 human Ig kappa chain signal peptide 2 MKYLLPTAAAGLLLLAAQPAMA77 AYRipr secretory signal MFRIFFTLLIIILIKKTSA78 AYCyRPA secretory signal MIIPFHKKFISFFQIVLVVLLLCRSINC79 PfCSS secretory signal MIKVLLAVLFILIKLENIIG156 HSV-2 gI + WR MPGRSLQGLAILGLWVCATGLVVR157 HSV-2 gI + L MPGRSLQGLAILGLWVCATGL158 HSV-1 gD + KY MGGAAARLGAVILFVVIVGLHGVRGKY159 HSV-2 gD + KYAL MGRLTSGVGTAALLVVAVGLRVVCAKYAL160 HSV-2 gD + KYALA MG RLTSG VGTAALLVVAVG LRVVCA KYALA161 HSV-2 gC signal MALGRV GLAVGLWGLLWVGVVVVLANApeptide162 IL2 signal peptide MRMQLLLLIALSLALVTNS163 HSV-1 gB + AP MHQGAPSWGRRWFWWALLGLTLGVLVASAAP164 HSV-2 gE + RTS MARGAGLVFFVGVWVVSCLAAAPRTS165 Ebola spike MGVTGILQLPRDRFKRTSFFLWVIILFQRTFSIP glycoprotein GP166 MHC Class II MAISGVPVLGFFIIAVLMSAQESWA167 Synthetic MDSKGSSQKGSRLLLLLVVSNLLLPQGVVG168 Japanese MWLVSLAIVTACAGAencephalitis JEV169 SARS-CoV-2 (short) MFVFLVLLPLVSSQC170 HSV-1 gD (strain MGGAAARLGAVILFVVIVGLHGVRGKYAKOS)Table 3: Exemplary polynucleotide sequences encoding secretory signalsSEQID Signal Sequence (Nucleotide)NO:80 HSV-1 gD SP wild-type AUGGGGGGGGCUGCCGCCAGGUUGGGGGCCGUGAUUUUGUUUGUCG UCAUAGUGGGCCUCCAUGGGGUCCGCAGCAAAUAU81 HSV-1 gD SP AUGGGAGGAGCCGCCGCCAGACUGGGAGCCGUGAUCCUGUUCGUGGU OptlO nt sequence GAUCGUGGGACUGCAUGGAGUGAGAAGCAAGUAC82 SARS-CoV-2-S AUGUUUGUGUUUCUUGUGCUGCUGCCUCUUGUGUCUUCUCAGUGUGU GAAUUUGACA83 human Ig heavy chain signal AUGGAUUGGAUUUGGAGAAUCCUGUUCCUCGUGGGAGCCGCUACAGG peptide (huSec) AGCCCACUCCCAGAUG84 human Ig heavy chain signal AUGGAGUUGGGACUGAGCUGGAUUUUCCUUUUGGCUAUUUUAAAAGG peptide 1 UGUCCAGUGUSEQID Signal Sequence (Nucleotide)NO:85 human Ig heavy chain signal AUGGAACUGGGGCUCCGCUGGGUUUUCCUUGUUGCUAUUUUAGAAGG peptide 2 UGUCCAGUGU86 human Ig heavy chain signal AUGAAACACCUGUGGUUCUUCCUCCUGCUGGUGGCAGCUCCCAGAUGG peptide 3 GUCCUGUCC87 human Ig heavy chain signal AUGGACUGGACCUGGAGGAUCCUCUUCUUGGUGGCAGCAGCAACAGG peptide 4 UGCCCACUCG88 human Ig heavy chain signal AUGGACUGGACCUGGAGGUUCCUCUUUGUGGUGGCAGCAGCUACAGG peptide 5 UGUCCAGUCC89 human Ig heavy chain signal AUGGAGUUUGGGCUGAGCUGGCUUUUUCUUGUGGCGAUUCUAAAAGG peptide 6 UGUCCAGUGU90 human Ig heavy chain signal AUGGAGUUUGGGCUGAGCUGGGUUUUCCUCGUUGCUCUUUUUAGAGG peptide 7 UGUCCAGUGU91 human Ig heavy chain signal AUGGACCUCCUGCACAAGAACAUGAAACACCUGUGGUUCUUCCUCCUC peptide 8 CUGGUGGCAGCUCCCAGAUGGGUGCUGUCC92 human Ig kappa chain signal AUGGACAUGAGGGUCCCUGCUCAGCUCCUGGGGCUCCUGCUGCUCUG peptide 1 GCUCUCAGGUGCCAGAUGU93 human Ig kappa chain signal AUGAAAUACCUAUUGCCUACGGCAGCCGCUGGAUUGUUAUUACUCGCG peptide 2 GCCCAGCCGGCCAUGGCCHSV-1 gD_RG AUGGGCGGAGCUGCUGCUAGACUGGGAGCCGUGAUCCUGUUCGUGGU 171Variation 1 UAUCGUGGGACUGCACGGCGUGCGGGGAHSV-1 gD_RG AUGGGCGGAGCUGCUGCUAGACUGGGAGCCGUGAUCCUGUUCGUGGU 172Variation 1.1 UAUCGUGGGACUGCAUGGCGUGCGGGGAHSV-1 gD_RG AUGGGCGGAGCUGCUGCUAGACUGGGAGCCGUGAUCCUGUUCGUGGU 173Variation 1.2 UAUCGUGGGACUGCAUGGCGUGCGGGGCHSV-1 gD_RG AUGGGCGGAGCUGCUGCUAGACUGGGAGCCGUGAUCCUGUUCGUGGU 174Variation 1.4 UAUCGUGGGACUGCACGGCGUCAGAGGCHSV-1 gD_RG AUGGGCGGAGCUGCUGCUAGACUGGGAGCCGUGAUCCUGUUCGUGGU 175Variation 1.4 UAUCGUGGGACUGCAUGGCGUCAGAGGCHSV-1 gD_RG AUGGGCGGAGCUGCUGCUAGACUGGGAGCCGUGAUCCUGUUCGUGGU 176Variation 1.5 UAUCGUGGGACUGCAUGGCGUGCGGGGUHSV-2 gl AUGCCAGGACGGAGCCUUCAGGGCUUGGCCAUACUGGGGCUUUGGGU 178Variation 2 GUGUGCAACCGGGUUGGUAGUUCGAHSV-2 gl AUGCCUGGCAGAUCUCUGCAAGGACUGGCCAUCCUCGGACUGUGGGU 179Variation 1 UUGCGCAACAGGCCUGGUUGUUAGAHSV-2 gl AUGCCUGGAAGAUCUCUGCAGGGACUGGCAAUUCUGGGACUGUGGGU 180Variation 3 GUGUGCAACAGGACUGGUGGUGAGAHSV-1 gD AUGGGAGGCGCAGCUGCCAGACUUGGUGCUGUGAUCCUGUUCGUGGU 181GAUUGUAGGGCUGCAU HSV-1 gD _add AUGGGAGGCGCAGCUGCCAGACUUGGUGCUGUGAUCCUGUUCGUGGU 182GAUUGUAGGGCUGCAUGGUGUCAGGGGCAAGUAU HSV-2 gD AUGGGCCGCCUGACCUCCGGCGUGGGCACCGCCGCCCUGCUGGUGGU 183GGCCGUGGGCCUGCGCGUGGUGUGCGCCSEQ ID Signal Sequence (Nucleotide)NO:HSV-2 gD2 AUGGGCAGACUGACAUCUGGCGUGGGAACAGCUGCUCUGCUGGUGGU 184Variant 1 UGCUGUGGGCCUGAGAGUCGUGUGUGCCHSV-2 gD AUGGGGAGACUCACAUCAGGCGUAGGAACCGCUGCCCUGUUGGUCGU 185Variant 2 GGCCGUUGGUCUGAGAGUUGUGUGUGCCHSV-2 gD AUGGGCAGACUGACCUCCGGCGUGGGCACCGCCGCCCUGCUGGUGGU 186Variant 3 GGCCGUGGGCCUGAGAGUGGUGUGCGCCHSV-2 gD KYA wt AUGGGCCGCCUGACCUCCGGCGUGGGCACCGCCGCCCUGCUGGUGGU 187GGCCGUGGGCCUGCGCGUGGUGUGCGCCAAGUACGCCC. Linkers

[0225] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more linkers. In some embodiments, a linker is or comprises 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids. In some embodiments, a linker is or comprises no more than about 30, 25, 20, 15, 10 or fewer amino acids. A linker can include any amino acid sequence and is not limited to any particular amino acids. In some embodiments, a linker comprises one or more glycine (G) amino acids. In some embodiments, a linker comprises one or more serine (S) amino acids. In some embodiments, a linker comprises a glycine-serine linker. A "glycine-serine linker" as used herein refers to a linker that comprises predominantly (e.g., 80% or more) glycine and serine amino acids. In some embodiments, a linker includes amino acids selected based on a cleavage predictor to generate highly-cleavable linkers.

[0226] In some embodiments, a linker is or comprises S-G4-S-G4-S. In some embodiments, a linker is or comprises GSPGSGSGS (SEQ ID NO: 111). In some embodiments, a linker is or comprises GGSGGGGSGG (SEQ ID NO: 112). In some embodiments, a linker is or comprises GGSGG (SEQ ID NO: 107). In some embodiments, a linker is one presented in Table 4. In some embodiments, a linker is or comprises a sequence as set forth in W02017 / 081082, which is incorporated herein by reference in its entirety (see SEQ ID NOs: 1509-1565, or a fragment or variant thereof).

[0227] In some embodiments, a Plasmodium polypeptide construct described herein comprises a linker between a C-terminal region or portion thereof and a transmembrane region. In some embodiments, a Plasmodium polypeptide construct described herein comprises a linker after a minor repeat sequence.

[0228] Exemplary linkers are provided in the following Table 4:Table 4: Exemplary linkersSEQ ID NO: Sequence (Amino Acid)97 SGGGGSGGGGS98 GSPGSGSGS99 GGSGGGGSGG100 GGS101 GGGS102 GGGGSGGGGSGGGGS103 AGNRVRRSVG104 GSGSGS105 GGSLGGGGSG106 SGG107 GGSGGD. Transmembrane Regions

[0229] In some embodiments, a Plasmodium polypeptide construct described herein includes a transmembrane region (also referred to as a "transmembrane domain"). In some embodiments, a transmembrane region is located at the N-terminus of a Plasmodium polypeptide construct. In some embodiments, a transmembrane region is located at the C-terminus of a Plasmodium polypeptide construct. In some embodiments, a transmembrane region is not located at the N-terminus or C-terminus of a Plasmodium polypeptide construct.

[0230] Transmembrane regions are known in the art, any of which can be utilized in a Plasmodium polypeptide construct described herein. In some embodiments, a transmembrane region comprises or is a transmembrane domain of Hemagglutinin (HA) of Influenza virus, Env of HIV-1, equine infectious anaemia virus (EIAV), murine leukaemia virus (MLV), mouse mammary tumor virus, G protein of vesicular stomatitis virus (VSV), Rabies virus, or a seven transmembrane domain receptor.

[0231] In some embodiments, a heterologous transmembrane region does not comprise a hemagglutin transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of a nonhuman transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of an HSV transmembrane region, e.g., an HSV-1 or HSV-2 transmembrane region. In some embodiments, an HSV transmembrane region comprises or consists of an HSV gD transmembrane region, e.g., comprising or consisting of an amino acid sequence of GLIAGAVGGSLLAALVICGIVYWMRRHTQKAPKRIRLPHIR (SEQ ID NO:94).

[0232] In some embodiments, a heterologous transmembrane region comprises or consists of a human transmembrane region. In some embodiments, a human transmembrane region comprises or consists of a human decay accelerating factor glycosylphosphatidylinositol (hDAF-GPI) anchor region. In some embodiments, an hDAF-GPI anchor region comprises or consists of an amino acid sequence of PNKGSGTTSGTTRLLSGHTCFTLTGLLGTLVTMGLLT (SEQ ID NO: 95).

[0233] In some embodiments, a transmembrane region comprises or consists of a Plasmodium transmembrane region. In some embodiments, a utilized transmembrane region is one that is normally associated with CSP in nature. In some embodiments, a Plasmodium transmembrane region comprises or consists of a Plasmodium CSP glycosylphosphatidylinositol (GPI) anchor region. In some embodiments, a Plasmodium CSP GPI anchor region is from Plasmodium falciparum. In some embodiments, a Plasmodium CSP GPI anchor region is from Plasmodium falciparum isolate 3D7 (SEQ ID NO: 96).

[0234] In some embodiments, a transmembrane region comprises Plasmodium TRAP transmembrane region. In some embodiments, a transmembrane region comprises Plasmodium Pl 13 transmembrane region.

[0235] In some embodiments, a utilized transmembrane region is a heterologous transmembrane region.

[0236] In some embodiments, a Plasmodium polypeptide construct described herein does not comprise a transmembrane region.

[0237] Exemplary transmembrane are provided in the following Table 5:Table 5: Exemplary transmembrane regionsSEQ Transmembrane Region Sequence (Amino Acid)ID NO:94 HSV-1 gD GLIAGAVGGSLLAALVICGIVYWMRRHTQKAPKRIRLPHIR95 hDAF-GPI anchor region PNKGSGTTSGTTRLLSGHTCFTLTGLLGTLVTMGLLT96 Plasmodium CSP GPI anchor CSSVFNVVNSSIGLIMLVLSFLFLNregionE. Multimerization Regions

[0238] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more multimerization regions (e.g., a heterologous multimerization region). In some embodiments, a heterologous multimerization region comprises a dimerization, trimerization or tetramerization region.

[0239] In some embodiments, a multimerization region is one described in W02017 / 081082, which is incorporated herein by reference in its entirety e.g., SEQ ID NOs: 1116-1167, or fragments or variants thereof). Exemplary trimerization and tetramerization regions include, but are not limited to, engineered leucine zippers, fibritin foldon domain from enterobacteria phage T4, GCN4pll, GCN4-pll, and p53.

[0240] In some embodiments, a provided Plasmodium polypeptide construct described herein is able to form a trimeric complex. For example, a provided Plasmodium polypeptide construct may comprise a multimerization region allowing formation of a multimeric complex, such as for example a trimeric complex of a Plasmodium polypeptide construct described herein. In some embodiments, a multimerization region allowing formation of a multimeric complex comprises a trimerization region, for example, a trimerization region described herein. In some embodiments, a Plasmodium polypeptide construct includes a T4-fibri tin-deri ved "foldon" trimerization region, for example, to increase its immunogenicity. In some embodiments, a Plasmodium polypeptide construct includes a multimerization region comprising or consisting of the amino acid sequence GYIPEAPRDGQAYVRKDGEWVLLSTFL (SEQ ID NO: 108).F. Self-Assembling Regions

[0241] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more selfassembling regions (e.g., a self-assembling nanoparticle region, e.g., a heterologous self-assembling nanoparticle region). In some embodiments, a self-assembling nanoparticle region is a ferritin region. In some embodiments, a ferritin region is from H. pylori. In some embodiments, a ferritin region comprises or consists of a sequence of: DIIKLLNEQVNKEMQSSNLYMSMSSWCYTHSLDGAGLFLFDHAAEEYEHAKKLIIFLNENNVPVQLTSISAPEHKFEGLTQIFQKAYE HEQHISESINNIVDHAIKSKDHATFNFLQWYVAEQHEEEVLFKDILDKIELIGNENHGLYLADQYVKGIAKSRKS (SEQ ID NO: 113).G. Tags

[0242] Those skilled in the art, reading the present disclosure, will appreciate that, in some embodiments, one or more tags may be used when designing and testing constructs (e.g., Plasmodium polypeptide constructs described herein) in certain contexts (e.g., in vitro, ex vivo, etc.). In some embodiments, one or more tags may be directly connected to a Plasmodium polypeptide construct through a peptide bond (e.g., at the 5'-end or 3'-end of a construct). In some embodiments, one or more tags may be connected to a Plasmodium polypeptide construct through one or more linkers (e.g., one or more linkers described herein). In some embodiments, one or more tags may be internally embedded within a Plasmodium polypeptide construct, wherein the one or more tags are directly connected to the construct through peptide bonds (e.g., at the 5'-end and 3'-end of the one or more tags). In some embodiments, one or more tags may be internally embedded within a Plasmodium polypeptide construct, wherein the one or more tags are connected to the construct through one or more linkers (e.g., one or more linkers described herein).

[0243] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more tags. In some embodiments, a Plasmodium polypeptide construct described herein includes one or more detection tags (e.g., Hi Bit tag, HA tag, etc.). In some embodiments, a Plasmodium polypeptide construct described herein includes a tag that comprises or consists of a HiBit tag. In some embodiments, a H i Bit tag has an amino acid sequence according to SEQ ID NO: 123.H. Embodiments of Plasmodium polypeptide constructs

[0244] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more Plasmodium Rh5 polypeptide regions or portions thereof as described above. Exemplary combinations of regions are described below.

[0245] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more regions or portions of a Rh5 polypeptide from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.1. Rh5 constructs

[0246] In some embodiments, the Plasmodium polypeptide construct described herein includes an ordered domain of Rh5 (e.g., N-terminal ordered domain or C-terminal ordered domain) from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7. In some embodiments, the Plasmodium polypeptide construct described herein includes an ordered domain of Rh5 (e.g., N-terminal ordered domain or C-terminal ordered domain) from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7 (referred to herein as a " Rh5 construct" or " Rh5 ordered domain").

[0247] In some embodiments, an ordered domain of Rh5 is or comprises an N-terminal ordered domain that is or comprises the amino acid sequence of positions 155-240 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 155-240 of SEQ ID NO: 1. In some embodiments, an N-terminal ordered domain comprises the amino acid sequence of positions 155-240 of SEQ ID NO: 1, or the amino acid sequence of positions 155-240 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0248] In some embodiments, an ordered domain of Rh5 comprises the amino acid sequence of positions 155-247 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 155-247 of SEQ ID NO: 1. In some embodiments, an N-terminal ordered domain comprises the amino acid sequence of positions 155-247 of SEQ ID NO: 1, or the amino acid sequence of positions 155-247 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0249] In some embodiments, an ordered domain of Rh5 is or comprises an N-terminal ordered domain that is or comprises the amino acid sequence of positions 145-240 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 145-240 of SEQ ID NO: 1. In some embodiments, an N-terminal ordered domain comprises the amino acid sequence of positions 145-240 of SEQ ID NO: 1, or the amino acid sequence of positions 145-240 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0250] In some embodiments, an ordered domain of Rh5 is or comprises an N-terminal ordered domain that is or comprises the amino acid sequence of positions 145-247 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 145-247 of SEQ ID NO: 1. In some embodiments, an N-terminal ordered domain comprises the amino acid sequence of positions 145-247 of SEQ ID NO: 1, or the amino acid sequence of positions 145-247 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0251] In some embodiments, an ordered domain of Rh5 is or comprises an N-terminal ordered domain that is or comprises the amino acid sequence of positions 140-247 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 140-247 of SEQ ID NO: 1. In some embodiments, an N-terminal ordered domain comprises the amino acid sequence of positions 140-247 of SEQ ID NO: 1, or the amino acid sequence of positions 140-247 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0252] In some embodiments, an ordered domain of Rh5 is or comprises a C-terminal ordered domain that is or comprises the amino acid sequence of positions 297-497 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 297-497 of SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises the amino acid sequence of positions 297-526 of SEQ ID NO: 1, or the amino acid sequence of positions 297-497 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0253] In some embodiments, an ordered domain of Rh5 is or comprises a C-terminal ordered domain that is or comprises the amino acid sequence of positions 297-506 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 297-506 of SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises the amino acid sequence of positions 297-506of SEQ ID NO: 1, or the amino acid sequence of positions 297-497 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0254] In some embodiments, an ordered domain of Rh5 is or comprises a C-terminal ordered domain that is or comprises the amino acid sequence of positions 297-526 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 297-526 of SEQ ID NO: 1. In some embodiments, a C-terminal ordered domain comprises the amino acid sequence of positions 297-526 of SEQ ID NO: 1, or the amino acid sequence of positions 297-526 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0255] In some embodiments, an N-terminal ordered domain of Rh5 can have the following structure:sequence corresponding to amino acids 155-240 of SEQ ID NO: 1 (155-247 N-terminal Ordered Domain); sequence corresponding to amino acids 155-240 of SEQ ID NO: 1 with a tyrosine at position 203 (155-240 N-terminal Ordered Domain-203Y);sequence corresponding to amino acids 155-240 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 214 (155-240 N-terminal Ordered Domain-Δ214gly);sequence corresponding to amino acids 155-240 of SEQ ID NO: 1 with a tyrosine at position 203 and a substitution that prevents glycosylation at position 214 (155-240 N-terminal Ordered Domain-203Y-Δ214gly);sequence corresponding to amino acids 155-240 of SEQ ID NO: 1 with a glutamine at position 214 (155-240 N-terminal Ordered Domain-N214Q);sequence corresponding to amino acids 155-240 of SEQ ID NO: 1 with a tyrosine at position 203 and a glutamine at position 214 (155-240 N-terminal Ordered Domain-203Y-N214Q);sequence corresponding to amino acids 155-247 of SEQ ID NO: 1 (155-247 N-terminal Ordered Domain); sequence corresponding to amino acids 155-247 of SEQ ID NO: 1 with a tyrosine at position 203 (155-247 N-terminal Ordered Domain-203Y);sequence corresponding to amino acids 155-247 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 214 (155-247 N-terminal Ordered Domain-Δ214gly);sequence corresponding to amino acids 155-247 of SEQ ID NO: 1 with a tyrosine at position 203 and a substitution that prevents glycosylation at position 214 (155-247 N-terminal Ordered Domain-203Y-Δ214gly);sequence corresponding to amino acids 155-247 of SEQ ID NO: 1 with a glutamine at position 214 (155-247 N-terminal Ordered Domain-N214Q);sequence corresponding to amino acids 155-247 of SEQ ID NO: 1 with a tyrosine at position 203 and a glutamine at position 214 (155-247 N-terminal Ordered Domain-203Y-N214Q);sequence corresponding to amino acids 145-240 of SEQ ID NO: 1 (145-240 N-terminal Ordered Domain); sequence corresponding to amino acids 145-240 of SEQ ID NO: 1 with a tyrosine at position 203 (145-240 N-terminal Ordered Domain-203Y);sequence corresponding to amino acids 145-240 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 214 (145-240 N-terminal Ordered Domain-Δ214gly);sequence corresponding to amino acids 145-240 of SEQ ID NO: 1 with a tyrosine at position 203 and a substitution that prevents glycosylation at position 214 (145-240 N-terminal Ordered Domain-203Y-Δ214gly);sequence corresponding to amino acids 145-240 of SEQ ID NO: 1 with a glutamine at position 214 (145-240 N-terminal Ordered Domain-N214Q);sequence corresponding to amino acids 145-240 of SEQ ID NO: 1 with a tyrosine at position 203 and a glutamine at position 214 (145-240 N-terminal Ordered Domain-203Y-N214Q);sequence corresponding to amino acids 145-247 of SEQ ID NO: 1 (145-247 N-terminal Ordered Domain); sequence corresponding to amino acids 145-247 of SEQ ID NO: 1 with a tyrosine at position 203 (145-247 N-terminal Ordered Domain-203Y);sequence corresponding to amino acids 145-247 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 214 (145-247 N-terminal Ordered Domain-Δ214gly);sequence corresponding to amino acids 145-247 of SEQ ID NO: 1 with a tyrosine at position 203 and a substitution that prevents glycosylation at position 214 (145-247 N-terminal Ordered Domain-203Y-Δ214gly);sequence corresponding to amino acids 145-247 of SEQ ID NO: 1 with a glutamine at position 214 (145-247 N-terminal Ordered Domain-N214Q);sequence corresponding to amino acids 145-247 of SEQ ID NO: 1 with a tyrosine at position 203 and a glutamine at position 214 (145-247 N-terminal Ordered Domain-203Y-N214Q);sequence corresponding to amino acids 145-247 of SEQ ID NO: 1 with an alanine at position 216 (145-247 N-terminal Ordered Domain-216A);sequence corresponding to amino acids 145-247 of SEQ ID NO: 1 with a tyrosine at position 203 and an alanine at position 216 (145-247 N-terminal Ordered Domain-203Y-216A);sequence corresponding to amino acids 145-247 of SEQ ID NO: 1 with a leucine at position 157, a glutamate at position 183, a tyrosine at position 203, an alanine at position 216, and a lysine at position 233 (145-247 N-terminal Ordered Domain-U57L-D183E-203Y-216A-A233K);sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 (140-247 N-terminal Ordered Domain); sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a tyrosine at position 203 (140-247 N-terminal Ordered Domain-203Y);sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 214 (140-247 N-terminal Ordered Domain-Δ214gly);sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a tyrosine at position 203 and a substitution that prevents glycosylation at position 214 (140-247 N-terminal Ordered Domain-203Y-Δ214gly);sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a glutamine at position 214 (140-247 N-terminal Ordered Domain-N214Q);sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a tyrosine at position 203 and a glutamine at position 214 (140-247 N-terminal Ordered Domain-203Y-N214Q);sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with an alanine at position 216 (140-247 N-terminal Ordered Domain-216A);sequence corresponding to amino acids 140-247 of SEQ ID NO: 1 with a tyrosine at position 203 and an alanine at position 216 (140-247 N-terminal Ordered Domain-203Y-216A);sequence corresponding to amino acids 297-497 of SEQ ID NO: 1 (297-497 C-terminal Ordered Domain); sequence corresponding to amino acids 297-497 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 297 (297-497 C-terminal Ordered Domain-A297gly);sequence corresponding to amino acids 297-497 of SEQ ID NO: 1 with a glutamine at position 297 (297-497 C-terminal Ordered Domain-N297Q);sequence corresponding to amino acids 297-506 of SEQ ID NO: 1 (297-506 C-terminal Ordered Domain); sequence corresponding to amino acids 297-506 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 297 (297-506 C-terminal Ordered Domain-A297gly);sequence corresponding to amino acids 297-506 of SEQ ID NO: 1 with a glutamine at position 297 (297-506 C-terminal Ordered Domain-N297Q);sequence corresponding to amino acids 297-506 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 297 (297-506 C-terminal Ordered Domain-A297gly);sequence corresponding to amino acids 297-506 of SEQ ID NO: 1 with an alanine at position 299 (297-506 C-terminal Ordered Domain-299A);sequence corresponding to amino acids 297-506 of SEQ ID NO: 1 with an alanine at position 299, an aspartate at position 304, an asparagine at position 312, a phenylalanine at position, a phenylalanine at position 314,a asparagine at position 316, an asparagine at position 330, an alanine at position 370, an asparagine at position 381, a lysine at position 384, a lysine at position 392, an asparagine at position 395, an glutamic acid at position 398, a lysine at position 458, a lysine at position 464, an alanine at position 467, and a leucine at position 505 (297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L);sequence corresponding to amino acids 297-526 of SEQ ID NO: 1 (297-526 C-terminal Ordered Domain); sequence corresponding to amino acids 297-526 of SEQ ID NO: 1 with a substitution that prevents glycosylation at position 297 (297-526 C-terminal Ordered Domain-A297gly); orsequence corresponding to amino acids 297-526 of SEQ ID NO: 1 with an alanine at position 299 (297-526 C-terminal Ordered Domain-299A).

[0256] In some embodiments, a Plasmodium polypeptide construct described herein includes two ordered domains of Rh5. In some embodiments, two ordered domains of Rh5 are from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7. In some embodiments, a Plasmodium polypeptide construct described herein includes an N-terminal ordered domain and a C-terminal ordered domain.

[0257] In some embodiments, a Plasmodium polypeptide construct comprises a Rh5 polypeptide or antigenic portion thereof that includes 2 ordered domains of Rh5. In some embodiments, an Rh5 portion of a malarial polypeptide construct includes two ordered domains of Rh5 that have the following structure:Rh5 ordered domain Constructs155-240 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-Δ214gly - 297-497 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-203Y-Δ214gly - 297-497 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain155-240 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain155-247 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y- 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-Δ214gly- 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y-Δ214gly- 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-N214Q- 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-Δ214gly - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-203Y-Δ214gly - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-Δ214gly - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y-Δ214gly - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-216A - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y-216A - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - 297-497 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-A214gly - 297-497 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-203Y-A214gly - 297-497 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-216A - 297-497 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-203Y-216A - 297-497 C-terminal Ordered Domain;-240 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-A214gly - 297-497 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y-A214gly - 297-497 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain-A297gly-240 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain-A297gly-A297gly-247 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain-A297gly-A297gly;-247 N-terminal Ordered Domain-203Y- 297-497 C-terminal Ordered Domain-A297gly-A297gly;-247 N-terminal Ordered Domain-A214gly- 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-A214gly- 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-N214Q- 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-A214gly - 297-497 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y-A214gly - 297-497 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-A214gly - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-A214gly - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-216A - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-216A - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - 297-497 C-terminal Ordered Domain-A297gly; -247 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-A214gly - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-A214gly - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-216A - 297-497 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-216A - 297-497 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-N297Q;-240 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-N297Q;-240 N-terminal Ordered Domain-A214gly - 297-506 C-terminal Ordered Domain-N297Q;-240 N-terminal Ordered Domain-203Y-A214gly - 297-506 C-terminal Ordered Domain-N297Q;-240 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-N297Q-240 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-N297Q-A297gly-247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-N297Q-A297gly;-247 N-terminal Ordered Domain-203Y- 297-506 C-terminal Ordered Domain-N297Q-A297gly;-247 N-terminal Ordered Domain-A214gly- 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-203Y-A214gly- 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-N214Q- 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-N297Q;-240 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-N297Q;-240 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-N297Q;-240 N-terminal Ordered Domain-A214gly - 297-506 C-terminal Ordered Domain-N297Q;-240 N-terminal Ordered Domain-203Y-A214gly - 297-506 C-terminal Ordered Domain-N297Q;-240 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-N297Q;-240 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-A214gly - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-203Y-A214gly - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-216A - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-203Y-216A - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - 297-506 C-terminal Ordered Domain-N297Q; -247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-A214gly - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-203Y-A214gly - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-216A - 297-506 C-terminal Ordered Domain-N297Q;-247 N-terminal Ordered Domain-203Y-216A - 297-506 C-terminal Ordered Domain-N297Q;-240 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-A214gly - 297-506 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y-A214gly - 297-506 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-A297gly-240 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-A297gly-A297gly -247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-A297gly-A297gly;-247 N-terminal Ordered Domain-203Y- 297-506 C-terminal Ordered Domain-A297gly-A297gly;-247 N-terminal Ordered Domain-A214gly- 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-A214gly- 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-N214Q- 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-A214gly - 297-506 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y-A214gly - 297-506 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-A214gly - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-A214gly - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-216A - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-216A - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - 297-506 C-terminal Ordered Domain-A297gly; -247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-A214gly - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-A214gly - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-216A - 297-506 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-216A - 297-506 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-A214gly - 297-506 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-203Y-A214gly - 297-506 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-299A-240 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-299A-A297gly-247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-299A-A297gly;-247 N-terminal Ordered Domain-203Y- 297-506 C-terminal Ordered Domain-299A-A297gly;-247 N-terminal Ordered Domain-A214gly- 297-506 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y-A214gly- 297-506 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-N214Q- 297-506 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-A214gly - 297-506 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-203Y-A214gly - 297-506 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-A214gly - 297-506 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y-A214gly - 297-506 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-216A - 297-506 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y-216A - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - 297-506 C-terminal Ordered Domain-299A; 140-247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-Δ214gly - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y-Δ214gly - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-216A - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y-216A - 297-506 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain-Δ214gly - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain-203Y-Δ214gly - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L155-240 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L-A297gly155-247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L-A297gly;155-247 N-terminal Ordered Domain-203Y- 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-247 N-terminal Ordered Domain-203Y-Δ214gly- 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-247 N-terminal Ordered Domain-N214Q- 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-Δ214gly - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-203Y-Δ214gly - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-Δ214gly - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-203Y-Δ214gly - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-216A - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-203Y-216A - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-Δ214gly - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-203Y-Δ214gly - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-216A - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-203Y-216A - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-203Y - 297-526 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-Δ214gly - 297-526 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-203Y-Δ214gly - 297-526 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-N214Q - 297-526 C-terminal Ordered Domain155-240 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain-A297gly155-247 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-203Y- 297-526 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- 297-526 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y-Δ214gly- 297-526 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-N214Q- 297-526 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain;-240 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain;-240 N-terminal Ordered Domain-203Y - 297-526 C-terminal Ordered Domain;-240 N-terminal Ordered Domain-A214gly - 297-526 C-terminal Ordered Domain;-240 N-terminal Ordered Domain-203Y-A214gly - 297-526 C-terminal Ordered Domain;-240 N-terminal Ordered Domain-N214Q - 297-526 C-terminal Ordered Domain;-240 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-203Y - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-A214gly - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-203Y-A214gly - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-N214Q - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-216A - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-203Y-216A - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - 297-526 C-terminal Ordered Domain; -247 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-203Y - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-A214gly - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-203Y-A214gly - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-N214Q - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-216A - 297-526 C-terminal Ordered Domain;-247 N-terminal Ordered Domain-203Y-216A - 297-526 C-terminal Ordered Domain;-240 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y - 297-526 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-A214gly - 297-526 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y-A214gly - 297-526 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-N214Q - 297-526 C-terminal Ordered Domain-A297gly-240 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain-A297gly-A297gly -247 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain-A297gly-A297gly;-247 N-terminal Ordered Domain-203Y- 297-526 C-terminal Ordered Domain-A297gly-A297gly;-247 N-terminal Ordered Domain-A214gly- 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-A214gly- 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-N214Q- 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y - 297-526 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-A214gly - 297-526 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y-A214gly - 297-526 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-N214Q - 297-526 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-A214gly - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-A214gly - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-N214Q - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-216A - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-216A - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - 297-526 C-terminal Ordered Domain-A297gly; -247 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-A214gly - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-A214gly - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-N214Q - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-216A - 297-526 C-terminal Ordered Domain-A297gly;-247 N-terminal Ordered Domain-203Y-216A - 297-526 C-terminal Ordered Domain-A297gly;-240 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-203Y - 297-526 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-A214gly - 297-526 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-203Y-A214gly - 297-526 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-N214Q - 297-526 C-terminal Ordered Domain-299A-240 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain-299A-A297gly-247 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain-299A-A297gly;-247 N-terminal Ordered Domain-203Y- 297-526 C-terminal Ordered Domain-299A-A297gly;-247 N-terminal Ordered Domain-A214gly- 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y-A214gly- 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-N214Q- 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-203Y - 297-526 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-A214gly - 297-526 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-203Y-A214gly - 297-526 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-N214Q - 297-526 C-terminal Ordered Domain-299A;-240 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-A214gly - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y-A214gly - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-N214Q - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-216A - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y-216A - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - 297-526 C-terminal Ordered Domain-299A; -247 N-terminal Ordered Domain - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-A214gly - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y-A214gly - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-N214Q - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-203Y-N214Q - 297-526 C-terminal Ordered Domain-299A;-247 N-terminal Ordered Domain-216A - 297-526 C-terminal Ordered Domain-299A; or140-247 N-terminal Ordered Domain-203Y-216A - 297-526 C-terminal Ordered Domain-299A.

[0258] In some embodiments, Plasmodium polypeptide construct includes an Rh5 polypeptide or antigenic portion thereof that includes 2 ordered domains of Rh5 and a linker. In some embodiments, an Rh5 portion of a malarial polypeptide has the following structure:Rh5 ordered domain with Linker Constructs155-240 N-terminal Ordered Domain - linker - 297-497 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-203Y - linker - 297-497 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-Δ214gly - linker - 297-497 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-497 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-N214Q - linker - 297-497 C-terminal Ordered Domain155-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-497 C-terminal Ordered Domain155-247 N-terminal Ordered Domain - linker - 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y- linker - 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-Δ214gly- linker - 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker - 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-N214Q- linker - 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain - linker - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-203Y - linker - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-Δ214gly - linker - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-N214Q - linker - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain - linker - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y - linker - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-Δ214gly - linker - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-N214Q - linker - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-216A - linker - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y-216A - linker - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - linker - 297-497 C-terminal Ordered Domain; 140-247 N-terminal Ordered Domain - linker - 297-497 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-203Y - linker - 297-497 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-Δ214gly - linker - 297-497 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-497 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-N214Q - linker - 297-497 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-497 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-216A - linker - 297-497 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-203Y-216A - linker - 297-497 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain - linker - 297-497 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-203Y - linker - 297-497 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-Δ214gly - linker - 297-497 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-497 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-N214Q - linker - 297-497 C-terminal Ordered Domain-A297gly155-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-497 C-terminal Ordered Domain-A297gly-A297gly155-247 N-terminal Ordered Domain - linker - 297-497 C-terminal Ordered Domain-A297gly-A297gly;155-247 N-terminal Ordered Domain-203Y- linker - 297-497 C-terminal Ordered Domain-A297gly-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker - 297-497 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker - 297-497 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-N214Q- linker - 297-497 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-497 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain - linker - 297-497 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-203Y - linker - 297-497 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-Δ214gly - linker - 297-497 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-497 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-N214Q - linker - 297-497 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain - linker - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y - linker - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-Δ214gly - linker - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-N214Q - linker - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-216A - linker - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y-216A - linker - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-U57L-D183E-203Y-216A-A233K - linker - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain - linker - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y - linker - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-Δ214gly - linker - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-N214Q - linker - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-216A - linker - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-216A - linker - 297-497 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-N297Q;155-240 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-N297Q;155-240 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-N297Q;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-N297Q;155-240 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-N297Q155-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-N297Q-A297gly 155-247 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-N297Q-A297gly;155-247 N-terminal Ordered Domain-203Y- linker - 297-506 C-terminal Ordered Domain-N297Q-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker - 297-506 C-terminal Ordered Domain-N297Q;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker - 297-506 C-terminal Ordered Domain-N297Q;155-247 N-terminal Ordered Domain-N214Q- linker - 297-506 C-terminal Ordered Domain-N297Q;155-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-N297Q;145-240 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-N297Q;145-240 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-N297Q;145-240 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-N297Q;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-N297Q;145-240 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-N297Q;145-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain-216A - linker - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain-203Y-216A - linker - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain-U57L-D183E-203Y-216A-A233K - linker - 297-506 C-terminal Ordered Domain-N297Q;140-247 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-N297Q;140-247 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-N297Q;140-247 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-N297Q;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-N297Q;140-247 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-N297Q;140-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-N297Q;140-247 N-terminal Ordered Domain-216A - linker - 297-506 C-terminal Ordered Domain-N297Q;140-247 N-terminal Ordered Domain-203Y-216A - linker - 297-506 C-terminal Ordered Domain-N297Q;155-240 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-A297gly155-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-A297gly-A297gly 155-247 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-A297gly-A297gly;155-247 N-terminal Ordered Domain-203Y- linker - 297-506 C-terminal Ordered Domain-A297gly-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker - 297-506 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker - 297-506 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-N214Q- linker - 297-506 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-216A - linker - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y-216A - linker - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-U57L-D183E-203Y-216A-A233K - linker - 297-506 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-216A - linker - 297-506 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-216A - linker - 297-506 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-299A155-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-299A-A297gly 155-247 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-299A-A297gly;155-247 N-terminal Ordered Domain-203Y- linker - 297-506 C-terminal Ordered Domain-299A-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker - 297-506 C-terminal Ordered Domain-299A;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker - 297-506 C-terminal Ordered Domain-299A;155-247 N-terminal Ordered Domain-N214Q- linker - 297-506 C-terminal Ordered Domain-299A;155-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-216A - linker - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-203Y-216A - linker - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-U57L-D183E-203Y-216A-A233K - linker - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-216A - linker - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y-216A - linker - 297-506 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L155-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L-A297gly155-247 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L-A297gly;155-247 N-terminal Ordered Domain-203Y- linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-247 N-terminal Ordered Domain-N214Q- linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-216A - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-203Y-216A - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-U57L-D183E-203Y-216A-A233K - linker - 297-506 C-terminal Ordered Domain- 299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L; 140-247 N-terminal Ordered Domain - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N- M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-203Y - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-N214Q - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-216A - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F- K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-203Y-216A - linker - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-203Y - linker - 297-526 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-Δ214gly - linker - 297-526 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-526 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-N214Q - linker - 297-526 C-terminal Ordered Domain155-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain-A297gly155-247 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-203Y- linker - 297-526 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker - 297-526 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker - 297-526 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-N214Q- linker - 297-526 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-203Y - linker - 297-526 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-Δ214gly - linker - 297-526 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-526 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-N214Q - linker - 297-526 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y - linker - 297-526 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-Δ214gly - linker - 297-526 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-526 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-N214Q - linker - 297-526 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-216A - linker - 297-526 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y-216A - linker - 297-526 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - linker - 297-526 C-terminal Ordered Domain; 140-247 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-203Y - linker - 297-526 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-Δ214gly - linker - 297-526 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-526 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-N214Q - linker - 297-526 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-216A - linker - 297-526 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-203Y-216A - linker - 297-526 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-203Y - linker - 297-526 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-Δ214gly - linker - 297-526 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-526 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-N214Q - linker - 297-526 C-terminal Ordered Domain-A297gly155-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain-A297gly-A297gly 155-247 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain-A297gly-A297gly;155-247 N-terminal Ordered Domain-203Y- linker - 297-526 C-terminal Ordered Domain-A297gly-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker - 297-526 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker - 297-526 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-N214Q- linker - 297-526 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-203Y - linker - 297-526 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-Δ214gly - linker - 297-526 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-526 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-N214Q - linker - 297-526 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y - linker - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-Δ214gly - linker - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-N214Q - linker - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-216A - linker - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y-216A - linker - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-U57L-D183E-203Y-216A-A233K - linker - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y - linker - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-Δ214gly - linker - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-N214Q - linker - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-216A - linker - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-216A - linker - 297-526 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain-203Y - linker - 297-526 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain-Δ214gly - linker - 297-526 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-526 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain-N214Q - linker - 297-526 C-terminal Ordered Domain-299A155-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain-299A-A297gly 155-247 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain-299A-A297gly;155-247 N-terminal Ordered Domain-203Y- linker - 297-526 C-terminal Ordered Domain-299A-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker - 297-526 C-terminal Ordered Domain-299A;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker - 297-526 C-terminal Ordered Domain-299A;155-247 N-terminal Ordered Domain-N214Q- linker - 297-526 C-terminal Ordered Domain-299A;155-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-203Y - linker - 297-526 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-Δ214gly - linker - 297-526 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-526 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-N214Q - linker - 297-526 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-203Y - linker - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-Δ214gly - linker - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-N214Q - linker - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-216A - linker - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-203Y-216A - linker - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-U57L-D183E-203Y-216A-A233K - linker - 297-526 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain - linker - 297-526 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y - linker - 297-526 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-Δ214gly - linker - 297-526 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker - 297-526 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-N214Q - linker - 297-526 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y-N214Q - linker - 297-526 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-216A - linker - 297-526 C-terminal Ordered Domain-299A; or140-247 N-terminal Ordered Domain-203Y-216A - linker - 297-526 C-terminal Ordered Domain-299A.

[0259] In some embodiments, Plasmodium polypeptide construct includes an Rh5 polypeptide or antigenic portions thereof that includes 2 ordered domains of Rh5 (e.g., an N-terminal ordered domain or a C-terminal ordered domain) and one or more disordered domains (e.g., a linker disordered domain or an N-terminal disordered domain). In some embodiments, a disordered domain is a linker disordered domain of Rh5. In some embodiments, a linker disordered domain comprises a sequence corresponding to amino acids 248-296 of SEQ ID NO: 1. In some embodiments, an N-terminal disordered domain (referred to interchangeably herein as an N-disordered domain)comprises a sequence corresponding to amino acids 26-139 of SEQ ID NO: 1. In some embodiments, an Rh5 portion comprises both a linker disordered domain and an N-terminal disordered domain of Rh5.

[0260] In some embodiments, an Rh5 portion of a malarial polypeptide has two ordered domains and one or two disordered domains of Rh5. In some embodiments, an Rh5 portion of a malarial polypeptide has the following structure:Rh5 ordered domains with Linker Disordered Domain Constructs155-240 N-terminal Ordered Domain - linker disordered domain - 297-497 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-497 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain; 155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain; 155-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain 155-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain 155-247 N-terminal Ordered Domain - linker disordered domain - 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y- linker disordered domain - 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-Δ214gly- linker disordered domain - 297-497 C-terminal Ordered Domain; 155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker disordered domain - 297-497 C-terminal Ordered Domain; 155-247 N-terminal Ordered Domain-N214Q- linker disordered domain - 297-497 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain; 145-240 N-terminal Ordered Domain - linker disordered domain - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-497 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain; 145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain; 145-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain; 145-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain; 145-247 N-terminal Ordered Domain - linker disordered domain - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain; 145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain; 145-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain; 145-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain; 145-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-497 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-497 C-terminal Ordered Domain; 145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - linker disordered domain - 297-497 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain - linker disordered domain - 297-497 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-497 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain; 140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain; 140-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain; 140-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain; 140-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-497 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-497 C-terminal Ordered Domain; 155-240 N-terminal Ordered Domain - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 155-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 155-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly 155-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly-A297gly155-247 N-terminal Ordered Domain - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly-A297gly;155-247 N-terminal Ordered Domain-203Y- linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-N214Q- linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 155-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 145-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 145-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 145-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 145-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 145-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 145-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 145-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 140-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 140-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly; 140-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-497 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 155-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 155-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;155-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q 155-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q-A297gly155-247 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q-A297gly; 155-247 N-terminal Ordered Domain-203Y- linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;155-247 N-terminal Ordered Domain-N214Q- linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 155-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;145-240 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 145-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 145-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;145-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 145-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 145-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 145-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 145-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 145-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;140-247 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 140-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 140-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;140-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 140-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;140-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q; 140-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-N297Q;155-240 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 155-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 155-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly 155-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly-A297gly155-247 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly-A297gly;155-247 N-terminal Ordered Domain-203Y- linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-N214Q- linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 155-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 145-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 145-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 145-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 145-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 145-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 145-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 145-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 140-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 140-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 140-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly; 140-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 155-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A 155-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-A297gly155-247 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-A297gly; 155-247 N-terminal Ordered Domain-203Y- linker disordered domain - 297-506 C-terminal Ordered Domain-299A-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker disordered domain - 297-506 C-terminal Ordered Domain-299A;155-247 N-terminal Ordered Domain-N214Q- linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 155-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 145-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 145-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 145-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 145-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 145-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 140-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 140-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-299A; 140-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L; 155-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L 155-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L-A297gly155-247 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L-A297gly;155-247 N-terminal Ordered Domain-203Y- linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L-A297gly; 155-247 N-terminal Ordered Domain-Δ214gly- linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L; 155-247 N-terminal Ordered Domain-N214Q- linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;155-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L; 145-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L; 145-247 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L; 145-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L; 145-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;145-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L; 145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L; 140-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L; 140-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L;140-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-506 C-terminal Ordered Domain-299A-M304F-K312N-L314F-K316N-M330N-S370A-S381N-T384K-L392K-T395N-N398E-R458K-N464K-S467A-F505L; 155-240 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-526 C-terminal Ordered Domain;155-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain; 155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain; 155-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain 155-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly155-247 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 155-247 N-terminal Ordered Domain-203Y- linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 155-247 N-terminal Ordered Domain-Δ214gly- linker disordered domain - 297-526 C-terminal Ordered Domain; 155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker disordered domain - 297-526 C-terminal Ordered Domain; 155-247 N-terminal Ordered Domain-N214Q- linker disordered domain - 297-526 C-terminal Ordered Domain;155-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain; 145-240 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-526 C-terminal Ordered Domain;145-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain; 145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain; 145-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain; 145-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain; 145-247 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-526 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain; 145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain; 145-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain; 145-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain; 145-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-526 C-terminal Ordered Domain;145-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-526 C-terminal Ordered Domain; 145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - linker disordered domain - 297-526 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-526 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain; 140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain; 140-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain; 140-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain; 140-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-526 C-terminal Ordered Domain;140-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-526 C-terminal Ordered Domain; 155-240 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 155-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 155-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly155-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly-A297gly155-247 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly-A297gly;155-247 N-terminal Ordered Domain-203Y- linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;155-247 N-terminal Ordered Domain-N214Q- linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 155-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 145-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 145-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;145-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 145-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 145-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 145-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 145-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 145-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 140-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 140-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly; 140-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;140-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-526 C-terminal Ordered Domain-A297gly;155-240 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 155-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 155-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;155-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-299A 155-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-299A-A297gly155-247 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain-299A-A297gly; 155-247 N-terminal Ordered Domain-203Y- linker disordered domain - 297-526 C-terminal Ordered Domain-299A-A297gly;155-247 N-terminal Ordered Domain-Δ214gly- linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 155-247 N-terminal Ordered Domain-203Y-Δ214gly- linker disordered domain - 297-526 C-terminal Ordered Domain-299A;155-247 N-terminal Ordered Domain-N214Q- linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 155-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-203Y - linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 145-240 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 145-240 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;145-240 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 145-240 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 145-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 145-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 145-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 145-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;145-247 N-terminal Ordered Domain-I157L-D183E-203Y-216A-A233K - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y - linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 140-247 N-terminal Ordered Domain-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-299A; 140-247 N-terminal Ordered Domain-203Y-Δ214gly - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-203Y-N214Q - linker disordered domain - 297-526 C-terminal Ordered Domain-299A;140-247 N-terminal Ordered Domain-216A - linker disordered domain - 297-526 C-terminal Ordered Domain-299A; or 140-247 N-terminal Ordered Domain-203Y-216A - linker disordered domain - 297-526 C-terminal Ordered Domain-299A.Rh5 ordered domains with N -Terminal Disordered (N -Disordered) Domain ConstructsN-Disordered Domain - 155-240 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 155-240 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 155-240 N-terminal Ordered Domain-Δ214gly - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 155-240 N-terminal Ordered Domain-203Y-Δ214gly - 297-497 C-terminal Ordered Domain; N-Disordered Domain - 155-240 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered DomainN-Disordered Domain - 155-240 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain N-Disordered Domain - 155-247 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 155-247 N-terminal Ordered Domain-203Y- 297-497 C-terminal Ordered Domain;N-Disordered Domain - 155-247 N-terminal Ordered Domain-Δ214gly- 297-497 C-terminal Ordered Domain;N-Disordered Domain - 155-247 N-terminal Ordered Domain-203Y-Δ214gly- 297-497 C-terminal Ordered Domain; N-Disordered Domain - 155-247 N-terminal Ordered Domain-N214Q- 297-497 C-terminal Ordered Domain;N-Disordered Domain - 155-247 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain; N-Disordered Domain - 145-240 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 145-240 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 145-240 N-terminal Ordered Domain-Δ214gly - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 145-240 N-terminal Ordered Domain-203Y-Δ214gly - 297-497 C-terminal Ordered Domain; N-Disordered Domain - 145-240 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 145-240 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain; N-Disordered Domain - 145-247 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 145-247 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 145-247 N-terminal Ordered Domain-Δ214gly - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 145-247 N-terminal Ordered Domain-203Y-Δ214gly - 297-497 C-terminal Ordered Domain; N-Disordered Domain - 145-247 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 145-247 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain; N-Disordered Domain - 145-247 N-terminal Ordered Domain-216A - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 145-247 N-terminal Ordered Domain-203Y-216A - 297-497 C-terminal Ordered Domain; N-Disordered Domain - 145-247 N-terminal Ordered Domain-U57L-D183E-203Y-216A-A233K - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 140-247 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 140-247 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 140-247 N-terminal Ordered Domain-Δ214gly - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 140-247 N-terminal Ordered Domain-203Y-Δ214gly - 297-497 C-terminal Ordered Domain; N-Disordered Domain - 140-247 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 140-247 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain; N-Disordered Domain - 140-247 N-terminal Ordered Domain-216A - 297-497 C-terminal Ordered Domain;N-Disordered Domain - 140-247 N-terminal Ordered Domain-203Y-216A - 297-497 C-terminal Ordered Domain; N-Disordered Domain - 155-240 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 155-240 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 155-240 N-terminal Ordered Domain-Δ214gly - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 155-240 N-terminal Ordered Domain-203Y-Δ214gly - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 155-240 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain-A297gly N-Disordered Domain - 155-240 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain-A297gly-A297glyN-Disordered Domain - 155-247 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain-A297gly-A297gly; N-Disordered Domain - 155-247 N-terminal Ordered Domain-203Y- 297-497 C-terminal Ordered Domain-A297gly-A297gly;N-Disordered Domain - 155-247 N-terminal Ordered Domain-Δ214gly- 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 155-247 N-terminal Ordered Domain-203Y-Δ214gly- 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 155-247 N-terminal Ordered Domain-N214Q- 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 155-247 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 145-240 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 145-240 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 145-240 N-terminal Ordered Domain-Δ214gly - 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 145-240 N-terminal Ordered Domain-203Y-Δ214gly - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 145-240 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 145-240 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 145-247 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 145-247 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 145-247 N-terminal Ordered Domain-Δ214gly - 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 145-247 N-terminal Ordered Domain-203Y-Δ214gly - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 145-247 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 145-247 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 145-247 N-terminal Ordered Domain-216A - 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 145-247 N-terminal Ordered Domain-203Y-216A - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 145-247 N-terminal Ordered Domain-U57L-D183E-203Y-216A-A233K - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 140-247 N-terminal Ordered Domain - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 140-247 N-terminal Ordered Domain-203Y - 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 140-247 N-terminal Ordered Domain-Δ214gly - 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 140-247 N-terminal Ordered Domain-203Y-Δ214gly - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 140-247 N-terminal Ordered Domain-N214Q - 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 140-247 N-terminal Ordered Domain-203Y-N214Q - 297-497 C-terminal Ordered Domain-A297gly;N-Disordered Domain - 140-247 N-terminal Ordered Domain-216A - 297-497 C-terminal Ordered Domain-A297gly; N-Disordered Domain - 140-247 N-terminal Ordered Domain-203Y-216A - 297-497 C-terminal Ordered Domain-A297gly;-n-N-Disordered Domain - 155-240 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-N297Q;N-Disordered Domain - 155-240 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 155-240 N-terminal Ordered Domain-Δ214gly - 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 155-240 N-terminal Ordered Domain-203Y-Δ214gly - 297-506 C-terminal Ordered Domain-N297Q;N-Disordered Domain - 155-240 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-N297Q N-Disordered Domain - 155-240 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-N297Q-A297glyN-Disordered Domain - 155-247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-N297Q-A297gly; N-Disordered Domain - 155-247 N-terminal Ordered Domain-203Y- 297-506 C-terminal Ordered Domain-N297Q-A297gly;N-Disordered Domain - 155-247 N-terminal Ordered Domain-Δ214gly- 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 155-247 N-terminal Ordered Domain-203Y-Δ214gly- 297-506 C-terminal Ordered Domain-N297Q;N-Disordered Domain - 155-247 N-terminal Ordered Domain-N214Q- 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 155-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-N297Q;N-Disordered Domain - 145-240 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-N297Q;N-Disordered Domain - 145-240 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 145-240 N-terminal Ordered Domain-Δ214gly - 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 145-240 N-terminal Ordered Domain-203Y-Δ214gly - 297-506 C-terminal Ordered Domain-N297Q;N-Disordered Domain - 145-240 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 145-240 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-N297Q;N-Disordered Domain - 145-247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-N297Q;N-Disordered Domain - 145-247 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 145-247 N-terminal Ordered Domain-Δ214gly - 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 145-247 N-terminal Ordered Domain-203Y-Δ214gly - 297-506 C-terminal Ordered Domain-N297Q;N-Disordered Domain - 145-247 N-terminal Ordered Domain-N214Q - 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 145-247 N-terminal Ordered Domain-203Y-N214Q - 297-506 C-terminal Ordered Domain-N297Q;N-Disordered Domain - 145-247 N-terminal Ordered Domain-216A - 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 145-247 N-terminal Ordered Domain-203Y-216A - 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 145-247 N-terminal Ordered Domain-U57L-D183E-203Y-216A-A233K - 297-506 C-terminal Ordered Domain-N297Q;N-Disordered Domain - 140-247 N-terminal Ordered Domain - 297-506 C-terminal Ordered Domain-N297Q;N-Disordered Domain - 140-247 N-terminal Ordered Domain-203Y - 297-506 C-terminal Ordered Domain-N297Q; N-Disordered Domain - 140-2...

Claims

CLAIMS1. A polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises two or more antigenic portions of Plasmodium Rh5, wherein the two or more antigenic portions of Plasmodium Rh5 comprise an N-terminal ordered domain and a C-terminal ordered domain, and wherein the N-terminal ordered domain and / or the C-terminal ordered domain comprise an amino acid substitution at one or more N-linked glycosylation sites, wherein the amino acid substitution at the one or more N-linked glycosylation sites comprises:(i) NX[T / S] to QX[T / S], or(ii) NX[T / S] to NXA.

2. The polyribonucleotide of claim 1, wherein:(i) the amino acid substitution at the one or more N-linked glycosylation sites of the N-terminal ordered domain comprises N214Q or T216A, wherein the amino acid numbering is relative to SEQ ID NO: 1;(ii) the N-terminal ordered domain comprises a C203Y mutation, wherein the amino acid numbering is relative to SEQ ID NO: 1;(iii) the N-terminal ordered domain comprises a L231T mutation, wherein the amino acid numbering is relative to SEQ ID NO: 1;(iv) the N-terminal ordered domain comprises:(a) I157L,(b) D183E,(c) A233K, or(d) any combination thereof,wherein the amino acid numbering is relative to SEQ ID NO: 1; and / or(v) the N-terminal ordered domain comprises:(a) I157L,(b) D183E, and(c) A233K,wherein the amino acid numbering is relative to SEQ ID NO: 1.

3. The polyribonucleotide of claim 1, wherein:(i) the amino acid substitution at the one or more N-linked glycosylation sites of the C-terminal ordered domain comprises N297Q or T299A, wherein the amino acid numbering is relative to SEQ ID NO: 1;(ii) the C-terminal ordered domain comprises:(a) L387T,(b) N398T, and(c) R458K,wherein the amino acid numbering is relative to SEQ ID NO: 1;(iii) the C-terminal ordered domain comprises:(a) M304F,(b) K312N,(c) L314F,(d) K316N,(e) M330N,(f) S370A,(g) S381N,(h) T384K,(i) L392K,(j) T395N,(k) N398E,(l) R458K,(m) N464K,(n) S467A,(o) F505L, or(p) any combination thereof,wherein the amino acid numbering is relative to SEQ ID NO: 1; and / or(iv) the C-terminal ordered domain comprises:(a) M304F,(b) K312N,(c) L314F,(d) K316N,(e) M330N,(f) S370A,(g) S381N,(h) T384K,(i) L392K,(j) T395N,(k) N398E,(l) R458K,(m) N464K,(n) S467A, and(o) F505L,wherein the amino acid numbering is relative to SEQ ID NO: 1.

4. The polyribonucleotide of any one of claims 1-3, wherein:(i) the N-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 33, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 or 34, and / or (ii) the C-terminal ordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 153, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 148, 149, 150, 152, 154 or 155.

5. The polyribonucleotide of any one of claims 1-4, wherein the one or more antigenic portions of Plasmodium Rh5 comprise a linking disordered domain, optionally comprising an amino acid substitution at one or more N-linked glycosylation sites said amino acid substitutions comprising:(i) NX[T / S] to QX[T / S],(ii) NX[T / S] to NXA,(iii) T286A, wherein the amino acid numbering is relative to SEQ ID NO: 1; or(iv) N284Q, wherein the amino acid numbering is relative to SEQ ID NO: 1.

6. The polyribonucleotide of claim 5, wherein the linking disordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 46, 48 or 49.

7. The polyribonucleotide of any one of claims 1-50, wherein the one or more antigenic portions of Plasmodium Rh5 comprise an N-terminal disordered domain, optionally comprising an amino acid substitution at one or more N-linked glycosylation sites said amino acid substitutions comprising:(i) NX[T / S] to QX[T / S],(ii) NX[T / S] to NXA,(iii) T40A, wherein the amino acid numbering is relative to SEQ ID NO: 1; or(iv) N38Q, wherein the amino acid numbering is relative to SEQ ID NO: 1.

8. The polyribonucleotide of claim 7, wherein the N-terminal disordered domain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 50, 51 or 52.

9. The polyribonucleotide of any one of claims 1-8, wherein the polypeptide comprises(i) one or more linkers optionally comprising SEQ ID NO: 107; and / or(ii) a secretory signal optionally comprising SEQ ID NO: 58.

10. The polyribonucleotide of any one of claims 1 to 9, wherein the polypeptide comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 143, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12,13, 14, 15, 16, 17, 18, 109, 110, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146 or 147.

11. An RNA construct comprising in 5' to 3' order:(i) a 5' UTR;(ii) a polyribonucleotide of any one of claims 1-10;(iii) a 3' UTR; and(iv) a polyA tail sequence.

12. A pharmaceutical composition comprising one or more polyribonucleotides of any one of claims 1 to 10 or RNA constructs of claim 11.

13. The pharmaceutical composition of claim 12, wherein the composition further comprises(i) lipid nanoparticles, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes,wherein the one or more polyribonucleotides are fully or partially encapsulated within the lipid nanoparticles, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes; and / or (ii) at least one pharmaceutically acceptable excipient.

14. A combination comprising:(i) a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide is a polyribonucleotide according to any one of claims 1-10; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, the second polypeptide comprises one or more Plasmodium T cell antigens, optionally selected from Plasmodium CSP, Pfs230, LISP-1, LISP-2, TRAP, LSAP2, LSA-3, LSA-l(a), LSA-l(b), UIS3, ETRAMP10.3, TLP, CelTos, PLP1, SPECT1, SPECT2, STARP, MAEBL, CLAMP, SALSA, SIAP1, LIMP, GEST, CyRPA, CSS, Ripr, TRAMP, GARP, PIESP2 and P113.

15. A method for treating or preventing a malaria infection, the method comprising administering the polyribonucleotide according to any one of claims 1-10, the RNA construct of claim 11, the composition of claim 12 or 13 or the combination of claim 14 to a subject.

16. The polyribonucleotide of any one of claims 1-10, the RNA construct of claim 11, the pharmaceutical composition of claim 12 or 13, or the combination of claim 14 for use in the treatment or prevention of a malaria infection comprising administering one or more doses of the pharmaceutical composition to a subject.

17. Use of the polyribonucleotide of any one of claims 1-10, the RNA construct of claim 11, the pharmaceutical composition of claim 12 or 13, or the combination of claim 14 in the treatment or prevention of a malaria infection.

18. Use of the polyribonucleotide of any one of claims 1-10, the RNA construct of claim 11, the pharmaceutical composition of claim 12 or 13 or the combination of claim 14 for the manufacture of a medicament for the treatment or prevention of a malaria infection.

19. A polypeptide encoded by a polyribonucleotide of any one of claims 1-10.

20. A host cell comprising a polyribonucleotide of any one of claims 1-10, an RNA construct of claim 11 or a polypeptide of claim 19.

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