Compositions and related methods for delivering plasmodium antigens

Abstract

This disclosure provides compositions (e.g., pharmaceutical compositions) and related technologies (e.g., their components and / or related methods) for the delivery of malaria protein antigens. In particular, this disclosure provides polyribonucleotides encoding malaria protein antigens.

Description

[Technical Field]

[0001] Cross-reference of related applications This application claims priority to U.S. Provisional Application No. 63 / 515,079, filed on 21 July 2023, which is incorporated herein by reference in its entirety. [Background technology]

[0002] Malaria is a mosquito-borne infectious disease caused by parasitic protozoa of the genus Plasmodium. According to the World Health Organization, an estimated 3.4 billion people in 92 countries are infected with the malaria parasite and are at risk of developing the disease. [Overview of the Initiative]

[0003] This disclosure provides techniques (e.g., compositions, methods, etc.) for the delivery of plasmodium antigens (also referred herein as “malaria antigens”). In one embodiment, polyribonucleotides encoding polypeptides comprising one or more plasmodium Rh5 entry complex polypeptides or antigenic moieties thereof are provided herein. In some embodiments, the moiety is the antigenic moiety.

[0004] In some embodiments, the polyribonucleotides provided herein are isolated polyribonucleotides. In some embodiments, the polyribonucleotides provided herein are manipulated polyribonucleotides. In some embodiments, the polyribonucleotides provided herein are codon-optimized polyribonucleotides.

[0005] In some embodiments, one or more plasmodium Rh5 entry complex antigens include (i) one or more plasmodium reticulocyte-binding protein homolog 5 (Rh5) polypeptides or antigenic moieties thereof; (ii) one or more plasmodium cysteine-rich protective antigen (CyRPA) polypeptides or antigenic moieties thereof; (iii) one or more plasmodium Rh5 interacting protein (Ripr) polypeptides or antigenic moieties thereof; (iv) one or more plasmodium P113 polypeptides or antigenic moieties thereof; (v) one or more plasmodium thrombospondin-associated apical merozoite protein (TRAMP) polypeptides or antigenic moieties thereof; or (vi) one or more plasmodium cysteine-rich small secretory protein (CSS) polypeptides or antigenic moieties thereof.

[0006] In some embodiments, the polyribonucleotide encodes a polypeptide comprising one or more plasmodium Rh5 entry complex polypeptides or their antigenic moieties. In some embodiments, the polyribonucleotide encodes a polypeptide comprising one or more plasmodium reticulocyte-binding protein homolog 5 (Rh5) polypeptides or their antigenic moieties.

[0007] In some embodiments, one or more plasmodium Rh5 intrusion complex polypeptides or their antigenic moieties include one or more antigenic moieties of plasmodium Rh5. In some embodiments, one or more plasmodium Rh5 antigenic moieties include one or more ordered domains of Rh5. In some embodiments, one or more plasmodium Rh5 antigenic moieties include two ordered domains of Rh5. In some embodiments, the two ordered domains of Rh5 are directly adjacent to each other. In some embodiments, one or more plasmodium Rh5 antigenic moieties include cysteine ​​at positions 203, 329, or both, numbered according to SEQ ID NO: 1. In some embodiments, one or more plasmodium Rh5 antigenic moieties include (i) the amino acid sequence according to SEQ ID NO: 118, 119, 124, or 126, (ii) the amino acid sequence according to SEQ ID NO: 120 or 128, or (iii) a combination thereof.

[0008] In some embodiments, one or more plasmodium Rh5 antigenic moieties contain tyrosine at position 203, position 329, or both, as numbered according to SEQ ID NO: 1. In some embodiments, one or more plasmodium Rh5 antigenic moieties contain (i) the amino acid sequence according to SEQ ID NO: 121, 122, 125, or 127, (ii) the amino acid sequence according to SEQ ID NO: 123 or 129, or (iii) a combination thereof. In some embodiments, one or more plasmodium Rh5 antigenic moieties contain tyrosine at position 203, as numbered according to SEQ ID NO: 1. In some embodiments, one or more plasmodium Rh5 antigenic moieties contain tyrosine at position 329, as numbered according to SEQ ID NO: 1.

[0009] In some embodiments, one or more plasmodium Rh5 antigenic moieties include one, two, three, or four N-linked glycosylation sites. In some embodiments, one or more plasmodium Rh5 antigenic moieties include two N-linked glycosylation sites. In some embodiments, one or more plasmodium Rh5 antigenic moieties include one or more amino acid substitutions at one or more N-linked glycosylation sites, the amino acid substitutions preventing glycosylation. In some embodiments, one or more plasmodium Rh5 antigenic moieties include substitution of NX[T / S] to QX[T / S] and / or substitution of NX[T / S] to NXA. In some embodiments, one or more plasmodium Rh5 antigenic moieties include (i) the amino acid sequence according to SEQ ID NO: 124, 125, 126, or 127, (ii) the amino acid sequence according to SEQ ID NO: 128 or 129, or (ii) a combination thereof.

[0010] In some embodiments, one or more plasmodium Rh5 antigenic moieties include an amino acid substitution that prevents glycosylation at position 214, numbered according to SEQ ID NO: 1. In some embodiments, one or more plasmodium Rh5 antigenic moieties include an amino acid substitution that prevents glycosylation at position 297, numbered according to SEQ ID NO: 1. In some embodiments, one or more plasmodium Rh5 antigenic moieties include an amino acid substitution at all N-linked glycosylation sites, and the amino acid substitution prevents glycosylation.

[0011] In some embodiments, one or more plasmodium Rh5 antigenic moieties include PMX cleavage sites. In some embodiments, one or more plasmodium Rh5 antigenic moieties include the amino acid sequence of SEQ ID NOs: 118, 121, 124, or 125. In some embodiments, one or more plasmodium Rh5 antigenic moieties do not include PMX cleavage sites.

[0012] In some embodiments, one or more plasmodium Rh5 antigenic moieties include the amino acid sequence of SEQ ID NOs. 119, 122, 126, or 127. In some embodiments, one or more plasmodium Rh5 antigenic moieties include (i) an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NOs. 130, (ii) an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NOs. 118, 119, 121, 122, 124, 125, 126, or 127, (iii) an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NOs. 131, (iv) an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NOs. 120, 123, 128, or 129, or (v) a combination thereof.

[0013] In some embodiments, one or more plasmodium Rh5 antigenic moieties include (i) an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 118, 119, 121, 122, 124, 125, 126, or 127; (ii) an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 120, 123, 128, or 129; or (iii) a combination thereof. In some embodiments, one or more plasmodium Rh5 antigenic moieties include (i) an amino acid sequence of SEQ ID NO: 121 or 122; and (ii) an amino acid sequence of SEQ ID NO: 120. In some embodiments, one or more plasmodium Rh5 antigenic moieties include (i) an amino acid sequence of SEQ ID NO: 125 or 127; and (ii) an amino acid sequence of SEQ ID NO: 120 or 128.

[0014] In some embodiments, one or more plasmodium Rh5 antigenic moieties include an amino acid sequence having at least 85% sequence identity with the amino acid sequence at positions 26-363 of SEQ ID NO: 94, 95, or 99. In some embodiments, one or more plasmodium Rh5 antigenic moieties include an amino acid sequence having at least 85% sequence identity with the amino acid sequence at positions 26-526 of SEQ ID NO: 89. In some embodiments, one or more plasmodium Rh5 antigenic moieties include the amino acid sequence at positions 26-363 of SEQ ID NO: 94. In some embodiments, one or more plasmodium Rh5 antigenic moieties include the amino acid sequence at positions 26-363 of SEQ ID NO: 95. In some embodiments, one or more plasmodium Rh5 antigenic moieties include the amino acid sequence at positions 26-363 of SEQ ID NO: 99. In some embodiments, one or more plasmodium Rh5 antigenic moieties include the amino acid sequence at positions 26-526 of SEQ ID NO: 89. In some embodiments, one or more plasmodium Rh5 antigenic moieties do not contain the disordered domain of Rh5. In some embodiments, one or more plasmodium Rh5 antigenic moieties include a PMX cleavage site. In some embodiments, the PMX cleavage site includes or consists of the amino acid sequence of NFLQ.

[0015] In some embodiments, one or more plasmodium Rh5 entry complex antigens comprise a plasmodium CyRPA polypeptide or its antigenic moiety.

[0016] In some embodiments, the plasmodium CyRPA antigenic moiety includes an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 133. In some embodiments, the plasmodium CyRPA antigenic moiety includes or consists of the amino acid sequence of SEQ ID NO: 133. In some embodiments, the plasmodium CyRPA antigenic moiety includes an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 169. In some embodiments, the plasmodium CyRPA antigenic moiety includes or consists of the amino acid sequence of SEQ ID NO: 169.

[0017] In some embodiments, the plasmodium CyRPA antigenic moiety includes one, two, or three N-linked glycosylation sites. In some embodiments, the plasmodium CyRPA antigenic moiety includes an amino acid substitution at one or more N-linked glycosylation sites, the amino acid substitution preventing glycosylation. In some embodiments, the plasmodium CyRPA antigenic moiety includes a substitution of NX[T / S] to QX[T / S]. In some embodiments, the plasmodium CyRPA antigenic moiety includes a substitution of NX[T / S] to NXA. In some embodiments, the plasmodium CyRPA antigenic moiety includes an amino acid substitution at all N-linked glycosylation sites, the amino acid substitution preventing glycosylation.

[0018] In some embodiments, the plasmodium CyRPA antigenic moiety contains asparagine at positions 145, 322, 338, or a combination thereof, as numbered according to SEQ ID NO: 3. In some embodiments, the plasmodium CyRPA antigenic moiety contains glutamine at positions 145, 322, 338, or a combination thereof, as numbered according to SEQ ID NO: 3. In some embodiments, the plasmodium CyRPA antigenic moiety contains glutamine at positions 145, 322, and 338, as numbered according to SEQ ID NO: 3. In some embodiments, the plasmodium CyRPA antigenic moiety contains or consists of the amino acid sequence of SEQ ID NO: 134. In some embodiments, the plasmodium CyRPA antigenic moiety contains or consists of the amino acid sequence of SEQ ID NO: 170.

[0019] In some embodiments, one or more plasmodium Rh5 entry complex antigens comprise a plasmodium P113 polypeptide or its antigenic moiety.

[0020] In some embodiments, the plasmodium P113 antigenic moiety includes an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 135. In some embodiments, the plasmodium P113 antigenic moiety includes or consists of the amino acid sequence of SEQ ID NO: 135.

[0021] In some embodiments, the plasmodium P113 antigenic moiety includes 1, 2, 3, 4, 5, 6, 7, or 8 N-linked glycosylation sites. In some embodiments, the plasmodium P113 antigenic moiety includes an amino acid substitution at one or more N-linked glycosylation sites, the amino acid substitution preventing glycosylation. In some embodiments, the plasmodium P113 antigenic moiety includes a substitution of NX[T / S] to QX[T / S]. In some embodiments, the plasmodium P113 antigenic moiety includes a substitution of NX[T / S] to NXA. In some embodiments, the plasmodium P113 antigenic moiety includes amino acid substitutions at all N-linked glycosylation sites, the amino acid substitution preventing glycosylation.

[0022] In some embodiments, the plasmodium P113 antigenic moiety contains asparagine at positions 207, 268, 317, 360, 661, 697, 779, 876, or combinations thereof, as numbered according to SEQ ID NO: 6. In some embodiments, the plasmodium P113 antigenic moiety contains glutamine at positions 207, 268, 317, 360, 661, 697, 779, 876, or combinations thereof, as numbered according to SEQ ID NO: 6. In some embodiments, the plasmodium P113 antigenic moiety contains glutamine at positions 207, 268, 317, 360, 661, 697, 779, and 876, as numbered according to SEQ ID NO: 6. In some embodiments, the plasmodium P113 antigenic moiety contains or consists of the amino acid sequence of SEQ ID NO: 136.

[0023] In some embodiments, one or more plasmodium Rh5 entry complex antigens comprise a plasmodium Ripr polypeptide or its antigenic moiety. In some embodiments, one or more plasmodium Rh5 entry complex antigens comprise an antigenic moiety of plasmodium Ripr.

[0024] In some embodiments, the plasmodium Ripr antigenic moiety includes or comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 178. In some embodiments, the plasmodium Ripr antigenic moiety includes or comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 177. In some embodiments, the plasmodium Ripr antigenic moiety includes or comprises the amino acid sequence of SEQ ID NO: 177.

[0025] In some embodiments, the plasmodium Ripr antigenic moiety includes or comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 176. In some embodiments, the plasmodium Ripr antigenic moiety includes or comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 175. In some embodiments, the plasmodium Ripr antigenic moiety includes or comprises the amino acid sequence of SEQ ID NO: 175.

[0026] In some embodiments, the plasmodium Ripr antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 174.

[0027] In some embodiments, the plasmodium Ripr antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 190.

[0028] In some embodiments, the plasmodium Ripr antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 171.

[0029] In some embodiments, the plasmodium Ripr antigenic moiety includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 N-linked glycosylation sites. In some embodiments, the plasmodium Ripr antigenic moiety includes an amino acid substitution at one or more N-linked glycosylation sites, the amino acid substitution preventing glycosylation. In some embodiments, the plasmodium Ripr antigenic moiety includes a substitution of NX[T / S] to QX[T / S]. In some embodiments, the plasmodium Ripr antigenic moiety includes an amino acid substitution at all N-linked glycosylation sites, the amino acid substitution preventing glycosylation.

[0030] In some embodiments, the plasmodium Ripr antigenic moiety contains asparagine at positions 646, 964, 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, the plasmodium Ripr antigenic moiety contains glutamine at positions 646, 964, 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, the plasmodium Ripr antigenic moiety contains glutamine at positions 646, 964, and 1021, as numbered according to SEQ ID NO: 2. In some embodiments, the plasmodium Ripr antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 190.

[0031] In some embodiments, the plasmodium Ripr antigenic moiety contains asparagine at positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, 1021, or combinations thereof, as numbered according to Sequence ID No. 2. In some embodiments, the plasmodium Ripr antigenic moiety contains glutamine at positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, 1021, or combinations thereof, as numbered according to Sequence ID No. 2. In some embodiments, the plasmodium Ripr antigenic moiety contains glutamine at positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021, numbered according to SEQ ID NO: 2. In some embodiments, the plasmodium Ripr antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 171.

[0032] In some embodiments, the plasmodium Ripr antigenic moiety includes a PMX cleavage site. In some embodiments, the PMX cleavage site includes or consists of the amino acid sequence of SMLE.

[0033] In some embodiments, one or more plasmodium Rh5 entry complex antigens comprise a plasmodium TRAMP polypeptide or its antigenic moiety.

[0034] In some embodiments, the plasmodium TRAMP antigenic moiety includes or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 179.

[0035] In some embodiments, the plasmodium TRAMP antigenic moiety includes 1, 2, 3, 4, 5, 6, 7, or 8 N-linked glycosylation sites. In some embodiments, the plasmodium TRAMP antigenic moiety includes an amino acid substitution at one or more N-linked glycosylation sites, the amino acid substitution preventing glycosylation. In some embodiments, the plasmodium TRAMP antigenic moiety includes a substitution of NX[T / S] to QX[T / S]. In some embodiments, the plasmodium TRAMP antigenic moiety includes an amino acid substitution at all N-linked glycosylation sites, the amino acid substitution preventing glycosylation.

[0036] In some embodiments, the plasmodium TRAMP antigenic moiety contains asparagine at positions 149, 195, 202, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, the plasmodium TRAMP antigenic moiety contains glutamine at positions 149, 195, 202, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, the plasmodium TRAMP antigenic moiety contains glutamine at positions 149, 195, and 202, as numbered according to SEQ ID NO: 4. In some embodiments, the plasmodium TRAMP antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 181.

[0037] In some embodiments, the plasmodium TRAMP antigenic moiety contains asparagine at positions 112, 149, 155, 170, 195, 202, 253, 305, or combinations thereof, as numbered according to SEQ ID NO: 4. In some embodiments, the plasmodium TRAMP antigenic moiety contains glutamine at positions 112, 149, 155, 170, 195, 202, 253, 305, or combinations thereof, as numbered according to SEQ ID NO: 4. In some embodiments, the plasmodium TRAMP antigenic moiety contains glutamine at positions 112, 149, 155, 170, 195, 202, 253, and 305, as numbered according to SEQ ID NO: 4. In some embodiments, the plasmodium TRAMP antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 180.

[0038] In some embodiments, the plasmodium TRAMP antigenic moiety includes a PMX cleavage site. In some embodiments, the PMX cleavage site includes or consists of the amino acid sequence of HFLQ. In some embodiments, the plasmodium TRAMP antigenic moiety includes a SUB2 cleavage site. In some embodiments, the SUB2 cleavage site includes or consists of the amino acid sequence of SEQ ID NO: 193.

[0039] In some embodiments, one or more plasmodium Rh5 entry complex antigens comprise a plasmodium CSS polypeptide or its antigenic moiety.

[0040] In some embodiments, the plasmodium CSS antigenic moiety comprises one or more cysteine-to-serine mutations. In some embodiments, the plasmodium CSS antigenic moiety comprises a C30S mutation, a C80S mutation, a C276S mutation, or a combination thereof. In some embodiments, the plasmodium CSS antigenic moiety comprises serine at positions 30 and 80, numbered according to SEQ ID NO: 5 or 214.

[0041] In some embodiments, the plasmodium CSS antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 194. In some embodiments, the plasmodium CSS antigenic moiety comprises or consists of an amino acid sequence of SEQ ID NO: 194. In some embodiments, the plasmodium CSS antigenic moiety comprises serine at positions 30 and 276, numbered according to SEQ ID NO: 5 or 214.

[0042] In some embodiments, the plasmodium CSS antigenic moiety includes or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 195.

[0043] In some embodiments, the plasmodium CSS antigenic moiety includes or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 182.

[0044] In some embodiments, the plasmodium CSS antigenic moiety includes 1, 2, 3, 4, 5, or 6 N-linked glycosylation sites. In some embodiments, the plasmodium CSS antigenic moiety includes an amino acid substitution at one or more N-linked glycosylation sites, the amino acid substitution preventing glycosylation. In some embodiments, the plasmodium CSS antigenic moiety includes a substitution of NX[T / S] to QX[T / S]. In some embodiments, the plasmodium CSS antigenic moiety includes an amino acid substitution at all N-linked glycosylation sites, the amino acid substitution preventing glycosylation.

[0045] In some embodiments, the plasmodium CSS antigenic moiety contains asparagine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, the plasmodium CSS antigenic moiety contains glutamine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 214.

[0046] In some embodiments, the plasmodium CSS antigenic moiety contains glutamine at positions 74 and 88, numbered according to SEQ ID NO: 5 or 214. In some embodiments, the plasmodium CSS antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 184.

[0047] In some embodiments, the plasmodium CSS antigenic moiety contains asparagine at positions 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 195. In some embodiments, the plasmodium CSS antigenic moiety contains glutamine at positions 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 195. In some embodiments, the plasmodium CSS antigenic moiety contains glutamine at positions 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 195. In some embodiments, the plasmodium CSS antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 185.

[0048] In some embodiments, the plasmodium CSS antigenic moiety contains asparagine at positions 74, 88, 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, the plasmodium CSS antigenic moiety contains glutamine at positions 74, 88, 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, the plasmodium CSS antigenic moiety contains glutamine at positions 74, 88, 192, 234, 261, 283, as numbered according to SEQ ID NO: 182. In some embodiments, the plasmodium CSS antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 183.

[0049] In some embodiments, the polypeptide comprises two or more plasmodium Rh5 entry complex polypeptides or their antigenic moieties. In some embodiments, the Rh5 entry complex polypeptide or its antigenic moiety comprises (i) one or more plasmodium Rh5 polypeptides or their antigenic moieties, (ii) one or more plasmodium CyRPA polypeptides or their antigenic moieties, (iii) one or more plasmodium Ripr polypeptides or their antigenic moieties, (iv) one or more plasmodium P113 polypeptides or their antigenic moieties, (v) one or more plasmodium TRAMP polypeptides or their antigenic moieties, and (vi) one or more plasmodium CSS polypeptides or their antigenic moieties.

[0050] In some embodiments, the polypeptide comprises one or more plasmodium Rh5 antigenic moieties, and further comprises (i) one or more plasmodium CyRPA polypeptides or antigenic moieties thereof, (ii) one or more plasmodium Ripr polypeptides or antigenic moieties thereof, (iii) one or more plasmodium P113 polypeptides or antigenic moieties thereof, (iv) one or more plasmodium TRAMP polypeptides or antigenic moieties thereof, or (v) one or more plasmodium CSS polypeptides or antigenic moieties thereof.

[0051] In some embodiments, the polypeptide comprises one or more plasmodium Rh5 antigenic moieties, wherein the polypeptide comprises (i) one or more plasmodium Rh5 polypeptides or antigenic moieties thereof and one or more plasmodium CyRPA polypeptides or antigenic moieties thereof; (ii) one or more plasmodium Rh5 polypeptides or antigenic moieties thereof and one or more plasmodium P113 polypeptides or antigenic moieties thereof; (iii) one or more plasmodium Rh5 polypeptides or antigenic moieties thereof and one or more plasmodium Ripr polypeptides or antigenic moieties thereof; (iv) one or more plasmodium Rh5 polypeptides or antigenic moieties thereof and one or more plasmodium TRAMP polypeptides or antigenic moieties thereof; or (v) one or more plasmodium Rh5 polypeptides or antigenic moieties thereof and one or more plasmodium CSS polypeptides or antigenic moieties thereof.

[0052] In some embodiments, the polypeptide includes a secretory signal. In some embodiments, the secretory signal is located at the N-terminus of the polypeptide.

[0053] In some embodiments, the secretion signal is a heterologous secretion signal. In some embodiments, the heterologous secretion signal includes or consists of a viral secretion signal. In some embodiments, the viral secretion signal includes or consists of an HSV secretion signal. In some embodiments, the HSV secretion signal includes or consists of an HSV-1 or HSV-2 secretion signal. In some embodiments, the HSV secretion signal includes or consists of an HSV glycoprotein D (gD) secretion signal. In some embodiments, the HSV gD secretion signal consists of the amino acid sequence according to SEQ ID NO: 42.

[0054] In some embodiments, the secretion signal includes or consists of a plasmodium secretion signal. In some embodiments, the secretion signal includes or consists of a plasmodium CyRPA secretion signal. In some embodiments, the secretion signal includes or consists of a plasmodium Ripr secretion signal. In some embodiments, the secretion signal includes or consists of a plasmodium CSS secretion signal. In some embodiments, the secretion signal includes or consists of the amino acid sequence of SEQ ID NOs: 186, 187, and 188.

[0055] In some embodiments, the polypeptide includes transmembrane regions. In some embodiments, the transmembrane regions include heterogeneous transmembrane regions.

[0056] In some embodiments, the transmembrane region includes or consists of a plasmodium transmembrane region. In some embodiments, the transmembrane region includes or consists of a P113 plasmodium transmembrane region. In some embodiments, the transmembrane region includes or consists of a TRAMP plasmodium transmembrane region.

[0057] In some embodiments, the heterologous transmembrane region includes or consists of a viral transmembrane region. In some embodiments, the heterologous transmembrane region includes or consists of an HSV transmembrane region. In some embodiments, the HSV transmembrane region includes or consists of an HSV-1 or HSV-2 transmembrane region. In some embodiments, the HSV transmembrane region includes or consists of an HSV gD transmembrane region. In some embodiments, the HSV gD transmembrane region consists of the amino acid sequence according to SEQ ID NO: 75.

[0058] In some embodiments, the polypeptide does not include a transmembrane region.

[0059] In some embodiments, the polypeptide includes a polymerizing domain. In some embodiments, the polymerizing domain is a trimerizing domain. In some embodiments, the trimerizing domain is the C-terminal domain of T4 fibrintin (e.g., the Foldon domain). In some embodiments, the Foldon domain includes or consists of the amino acid sequence given by SEQ ID NO: 78.

[0060] In some embodiments, the polypeptide includes a self-assembling nanoparticle domain. In some embodiments, the self-assembling nanoparticle domain is a ferritin domain. In some embodiments, the ferritin domain is derived from H. pylori. In some embodiments, the ferritin domain includes or consists of the sequence given by SEQ ID NO: 88.

[0061] In some embodiments, the polypeptide comprises one or more linkers. In some embodiments, the polypeptide comprises one or more glycine-serine linkers. In some embodiments, one or more linkers comprises or consist of the amino acid sequence according to SEQ ID NO: 80. In some embodiments, one or more linkers comprises or consist of the amino acid sequence according to SEQ ID NO: 83. In some embodiments, one or more linkers comprises or consist of the amino acid sequence according to SEQ ID NO: 84. In some embodiments, one or more linkers comprises or consist of the amino acid sequence according to SEQ ID NO: 86. In some embodiments, one or more linkers comprises or consist of the amino acid sequence according to SEQ ID NO: 137. In some embodiments, one or more linkers comprises or consist of the amino acid sequence according to SEQ ID NO: 138.

[0062] In some embodiments, the polypeptide includes a linker after the amino acid sequence of one or more plasmodium Rh5 entry complex polypeptides or their antigenic moieties.

[0063] In some embodiments, Plasmodium is Plasmodium falciparum. In some embodiments, Plasmodium falciparum isolate 3D7.

[0064] In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or their antigenic moieties are one or more Plasmodium falciparum Rh5 invasion complex polypeptides or their antigenic moieties. In some embodiments, one or more Plasmodium Rh5 polypeptides or their antigenic moieties are one or more Plasmodium falciparum Rh5 antigenic moieties. In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or their antigenic moieties are Plasmodium falciparum CyRPA polypeptides or their antigenic moieties. In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or their antigenic moieties are Plasmodium falciparum P113 polypeptides or their antigenic moieties. In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or their antigenic moieties are Plasmodium falciparum Ripr polypeptides or their antigenic moieties. In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or their antigenic moieties are Plasmodium falciparum TRAMP polypeptides or their antigenic moieties. In some embodiments, one or more Plasmodium Rh5 invasion complex polypeptides or their antigenic moieties are Plasmodium falciparum CSS polypeptides or their antigenic moieties.

[0065] Polyribonucleotides encoding polypeptides containing secretory signaling and two plasmodium Rh5 ordered domains are provided herein.

[0066] In some embodiments, the polypeptide comprises (i) a secretion signal, (ii) two plasmodium Rh5 ordered domains, (iii) a linker, and (iv) a polymerizing domain. In some embodiments, the polypeptide comprises (i) a secretion signal, (ii) two plasmodium Rh5 ordered domains, (iii) a linker, and (iv) a transmembrane region. In some embodiments, the polypeptide comprises (i) a secretion signal, (ii) two plasmodium Rh5 ordered domains, (iii) a linker, and (iv) a self-assembling nanoparticle domain.

[0067] In some embodiments, the secretion signal is a viral secretion signal. In some embodiments, the secretion signal is an HSV glycoprotein D (gD) secretion signal, and optionally the HSV gD secretion signal consists of the amino acid sequence of SEQ ID NO: 42.

[0068] In some embodiments, the polypeptide does not contain a plasmodium Rh5 disordered domain. In some embodiments, two plasmodium Rh5 ordered domains are directly adjacent.

[0069] In some embodiments, the polypeptide includes the C203Y substitution numbered according to SEQ ID NO: 1.

[0070] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 94.

[0071] In some embodiments, the linker comprises or consists of a glycine-serine linker. In some embodiments, the linker comprises or consists of the amino acid sequence according to SEQ ID NO: 86.

[0072] In some embodiments, the multimerization domain is the C-terminal domain of T4 fibrintin (e.g., the Foldon domain). In some embodiments, the Foldon domain comprises or consists of the amino acid sequence given by SEQ ID NO: 78.

[0073] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 90.

[0074] In some embodiments, the transmembrane region is a heterogeneous transmembrane region. In some embodiments, the transmembrane region is a viral transmembrane region. In some embodiments, the viral transmembrane region is an HSV gD transmembrane region. In some embodiments, the HSV gD transmembrane region contains or consists of the amino acid sequence of SEQ ID NO: 75.

[0075] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 92.

[0076] In some embodiments, the linker comprises or consists of a glycine-serine linker. In some embodiments, the linker comprises or consists of the amino acid sequence according to SEQ ID NO: 137.

[0077] In some embodiments, the self-assembling nanoparticle domain is a ferritin domain. In some embodiments, the ferritin domain includes or consists of the sequence given by Sequence ID No. 88.

[0078] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 96.

[0079] In some embodiments, the ordered domain of plasmodium Rh5 includes one or more N-linked glycosylation sites.

[0080] In some embodiments, the ordered domain of plasmodium Rh5 includes a single NX[T / S] to QX[T / S] substitution numbered according to Sequence ID No. 1, and optionally, the substitution is an N214Q substitution.

[0081] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 99.

[0082] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 98.

[0083] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 101.

[0084] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 100.

[0085] In some embodiments, the ordered domain of plasmodium Rh5 includes two NX[T / S] to QX[T / S] substitutions, optionally including N214Q and N297Q substitutions. In some embodiments, the ordered domain of plasmodium Rh5 includes amino acid substitutions at all N-linked glycosylation sites, where the amino acid substitutions prevent glycosylation.

[0086] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 95.

[0087] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 91.

[0088] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 93.

[0089] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 97.

[0090] Also provided herein are polyribonucleotides encoding polypeptides comprising (i) a secretory signal, (ii) a plasmodium Rh5 N-terminal disordered domain, (iii) a plasmodium Rh5 ordered domain, (iv) a plasmodium Rh5 linked disordered domain, and (v) a plasmodium Rh5 C-terminal ordered domain.

[0091] In some embodiments, the secretion signal is a heterologous secretion signal. In some embodiments, the heterologous secretion signal is a viral secretion signal. In some embodiments, the viral secretion signal is an HSV glycoprotein D (gD) secretion signal. In some embodiments, the HSV gD secretion signal includes or consists of the amino acid sequence of SEQ ID NO: 42.

[0092] In some embodiments, the polypeptide comprises one or more N-linked glycosylation sites. In some embodiments, the polypeptide comprises amino acid substitutions in one or more N-linked glycosylation sites, the amino acid substitutions preventing glycosylation, and optionally, the polypeptide comprises amino acid substitutions in all N-linked glycosylation sites.

[0093] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 89. In some embodiments, the polypeptide is or comprises the amino acid sequence of SEQ ID NO: 89.

[0094] This disclosure further provides (i) secretory signals and (ii) polyribonucleotides encoding a polypeptide comprising a plasmodium CyRPA polypeptide or its antigenic moiety.

[0095] In some embodiments, the polypeptide comprises (i) a secretory signal, (ii) a plasmodium CyRPA polypeptide or its antigenic moiety, (iii) a linker, and (iv) a polymerizing domain.

[0096] In some embodiments, the polypeptide comprises (i) a secretory signal, (ii) a plasmodium CyRPA polypeptide or its antigenic moiety, and (iii) a transmembrane region.

[0097] In some embodiments, the polypeptide comprises (i) a secretory signal, (ii) a plasmodium CyRPA polypeptide or its antigenic moiety, (iii) a linker, and (iv) a transmembrane region.

[0098] In some embodiments, the secretion signal is a heterologous secretion signal. In some embodiments, the heterologous secretion signal is a viral secretion signal. In some embodiments, the viral secretion signal is an HSV glycoprotein D (gD) secretion signal. In some embodiments, the HSV gD secretion signal includes or consists of the amino acid sequence of SEQ ID NO: 42.

[0099] In some embodiments, the secretion signal is a plasmodium secretion signal. In some embodiments, the plasmodium secretion signal is a plasmodium CyRPA secretion signal. In some embodiments, the plasmodium CyRPA secretion signal includes or consists of the amino acid sequence of SEQ ID NO: 187. In some embodiments, the plasmodium CyRPA polypeptide or its antigenic moiety includes or consists of an amino acid sequence having at least 85% sequence identity with the sequence of SEQ ID NO: 133.

[0100] In some embodiments, the plasmodium CyRPA polypeptide or its antigenic moiety comprises or consists of SEQ ID NO: 169 or 133. In some embodiments, the plasmodium CyRPA polypeptide or its antigenic moiety comprises or consists of amino acids 29 to 362 of SEQ ID NO: 169 or 133.

[0101] In some embodiments, the plasmodium CyRPA polypeptide or its antigenic moiety includes one or more N-linked glycosylation sites. In some embodiments, the plasmodium CyRPA polypeptide or its antigenic moiety includes one or more amino acid substitutions at the N-linked glycosylation sites, the amino acid substitutions preventing glycosylation.

[0102] In some embodiments, the plasmodium CyRPA polypeptide or its antigenic moiety includes substitution of NX[T / S] with QX[T / S] and / or substitution of NX[T / S] with NXA. In some embodiments, the plasmodium CyRPA polypeptide or its antigenic moiety includes N145Q, N322Q, N338Q, or a combination thereof. In some embodiments, the plasmodium CyRPA polypeptide or its antigenic moiety includes N145Q, N322Q, and N338Q.

[0103] In some embodiments, the linker comprises a glycine-serine linker. In some embodiments, the linker comprises or consists of the amino acid sequence according to SEQ ID NO: 86. In some embodiments, the linker comprises or consists of the amino acid sequence according to SEQ ID NO: 138.

[0104] In some embodiments, the multimerization domain is the C-terminal domain (Foldon domain) of T4 fibrintin. In some embodiments, the Foldon domain contains or consists of the amino acid sequence given by SEQ ID NO: 78.

[0105] In some embodiments, the transmembrane region is a heterogeneous transmembrane region. In some embodiments, the transmembrane region is a viral transmembrane region. In some embodiments, the viral transmembrane region is an HSV gD transmembrane region. In some embodiments, the HSV gD transmembrane region contains or consists of the amino acid sequence of SEQ ID NO: 75.

[0106] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 140.

[0107] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 141.

[0108] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 142.

[0109] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 143.

[0110] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 144.

[0111] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 198.

[0112] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 199.

[0113] Further provided are polyribonucleotides encoding a polypeptide comprising (i) a secretory signal, (ii) a plasmodium P113 polypeptide or its antigenic moiety, (iii) a linker, and (iv) a transmembrane region. In some embodiments, the polypeptide comprises (i) a secretory signal, (ii) a plasmodium P113 polypeptide or its antigenic moiety, (iii) a linker, and (iv) a transmembrane region.

[0114] In some embodiments, the secretion signal is a heterologous secretion signal. In some embodiments, the heterologous secretion signal is a viral secretion signal. In some embodiments, the viral secretion signal is an HSV glycoprotein D (gD) secretion signal. In some embodiments, the HSV gD secretion signal includes or consists of the amino acid sequence of SEQ ID NO: 42.

[0115] In some embodiments, the linker comprises a glycine-serine linker. In some embodiments, the linker comprises or consists of the amino acid sequence given by SEQ ID NO: 86.

[0116] In some embodiments, the transmembrane region is a heterogeneous transmembrane region. In some embodiments, the transmembrane region is a viral transmembrane region. In some embodiments, the viral transmembrane region is an HSV gD transmembrane region. In some embodiments, the HSV gD transmembrane region contains or consists of the amino acid sequence of SEQ ID NO: 75.

[0117] In some embodiments, the plasmodium P113 polypeptide or its antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 135.

[0118] In some embodiments, the plasmodium P113 polypeptide or its antigenic moiety includes one or more N-linked glycosylation sites. In some embodiments, the plasmodium P113 polypeptide or its antigenic moiety includes one or more amino acid substitutions of the N-linked glycosylation sites, the amino acid substitutions preventing glycosylation. In some embodiments, the plasmodium P113 polypeptide or its antigenic moiety includes one or more substitutions of NX[T / S] to QX[T / S] and / or substitutions of NX[T / S] to NXA.

[0119] In some embodiments, the plasmodium P113 antigenic moiety contains asparagine at positions 207, 268, 317, 360, 661, 694, 779, 876, or combinations thereof, as numbered according to Sequence ID No. 6. In some embodiments, the plasmodium P113 antigenic moiety contains glutamine at positions 207, 268, 317, 360, 661, 694, 779, 876, or combinations thereof, as numbered according to Sequence ID No. 6. In some embodiments, the plasmodium P113 antigenic moiety contains glutamine at positions 207, 268, 317, 360, 661, 694, 779, and 876, as numbered according to Sequence ID No. 6.

[0120] In some embodiments, the plasmodium P113 polypeptide or its antigenic moiety comprises or consists of the amino acid sequence given by SEQ ID NO: 136.

[0121] (i) secretory signals and (ii) polyribonucleotides encoding a polypeptide comprising a plasmodium Ripr polypeptide or its antigenic moiety are further provided.

[0122] In some embodiments, the polypeptide comprises (i) a secretory signal, (ii) a plasmodium Ripr polypeptide or its antigenic moiety, and (iii) a transmembrane region.

[0123] In some embodiments, the polypeptide comprises (i) a secretory signal, (ii) a plasmodium Ripr polypeptide or its antigenic moiety, (iii) a linker, and (iv) a transmembrane region.

[0124] In some embodiments, the secretion signal is a plasmodium secretion signal. In some embodiments, the plasmodium secretion signal is a plasmodium Ripr secretion signal. In some embodiments, the plasmodium Ripr secretion signal includes or consists of the amino acid sequence of SEQ ID NO: 186.

[0125] In some embodiments, the secretion signal is a viral secretion signal. In some embodiments, the secretion signal is an HSV glycoprotein D (gD) secretion signal. In some embodiments, the HSV gD secretion signal includes or consists of the amino acid sequence of SEQ ID NO: 42.

[0126] In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety Ripr comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 171.

[0127] In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 N-linked glycosylation sites. In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety comprises an amino acid substitution at one or more N-linked glycosylation sites, the amino acid substitution preventing glycosylation.

[0128] In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety includes a substitution of NX[T / S] to QX[T / S]. In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety includes amino acid substitutions at all N-linked glycosylation sites, and the amino acid substitutions prevent glycosylation.

[0129] In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety contains asparagine at positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, 1021, or combinations thereof, as numbered according to SEQ ID NO: 171. In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety contains glutamine at positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, 1021, or combinations thereof, as numbered according to SEQ ID NO: 171. In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety contains glutamine at positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021, numbered according to SEQ ID NO: 171. In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 172.

[0130] In some embodiments, the linker comprises a glycine-serine linker. In some embodiments, the linker comprises or consists of the amino acid sequence given by SEQ ID NO: 138.

[0131] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 145.

[0132] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 147.

[0133] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 149.

[0134] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 151.

[0135] In some embodiments, the transmembrane region is a heterogeneous transmembrane region. In some embodiments, the heterogeneous transmembrane region includes or consists of a viral transmembrane region. In some embodiments, the heterogeneous transmembrane region includes or consists of an HSV transmembrane region. In some embodiments, the HSV transmembrane region includes or consists of an HSV gD transmembrane region. In some embodiments, the HSV gD transmembrane region includes or consists of the amino acid sequence according to SEQ ID NO: 75.

[0136] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 146.

[0137] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 200.

[0138] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 148.

[0139] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 201.

[0140] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 150.

[0141] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 202.

[0142] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 152.

[0143] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 203.

[0144] In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 178.

[0145] In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 177.

[0146] In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 176.

[0147] In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 175.

[0148] In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 174.

[0149] The plasmodium Ripr polypeptide or its antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 190. In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety comprises or consists of the amino acid sequence of SEQ ID NO: 190.

[0150] In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety includes an amino acid substitution at one or more N-linked glycosylation sites, the amino acid substitution preventing glycosylation. In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety includes a substitution of NX[T / S] to QX[T / S]. In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety includes amino acid substitutions at all N-linked glycosylation sites, the amino acid substitution preventing glycosylation.

[0151] In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety contains asparagine at positions 646, 964, 1021, or a combination thereof, as numbered according to SEQ ID NO: 190. In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety contains glutamine at positions 646, 964, 1021, or a combination thereof, as numbered according to SEQ ID NO: 190. In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety contains glutamine at positions 646, 964, and 1021, as numbered according to SEQ ID NO: 190. In some embodiments, the plasmodium Ripr polypeptide or its antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 173.

[0152] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 153.

[0153] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 204.

[0154] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 154.

[0155] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 205.

[0156] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 155.

[0157] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 206.

[0158] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 156.

[0159] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 207.

[0160] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 157.

[0161] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 208.

[0162] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 158.

[0163] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 209.

[0164] (i) secretory signals, (ii) polyribonucleotides encoding a plasmodium TRAMP polypeptide or a polypeptide containing an antigenic moiety thereof are further provided.

[0165] In some embodiments, the secretion signal is a viral secretion signal. In some embodiments, the secretion signal is an HSV glycoprotein D (gD) secretion signal. In some embodiments, the HSV gD secretion signal includes or consists of the amino acid sequence of SEQ ID NO: 42.

[0166] In some embodiments, the plasmodium TRAMP polypeptide or its antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 179.

[0167] In some embodiments, the plasmodium trap polypeptide or its antigenic moiety comprises 1, 2, 3, 4, 5, 6, 7, or 8 N-linked glycosylation sites. In some embodiments, the plasmodium trap polypeptide or its antigenic moiety comprises amino acid substitutions at one or more N-linked glycosylation sites, the amino acid substitutions preventing glycosylation. In some embodiments, the plasmodium trap polypeptide or its antigenic moiety comprises a substitution of NX[T / S] to QX[T / S]. In some embodiments, the plasmodium trap polypeptide or its antigenic moiety comprises amino acid substitutions at all N-linked glycosylation sites, the amino acid substitutions preventing glycosylation.

[0168] In some embodiments, the plasmodium TRAMP polypeptide or its antigenic moiety contains asparagine at positions 149, 195, 202, or a combination thereof, as numbered according to SEQ ID NO: 179. In some embodiments, the plasmodium TRAMP polypeptide or its antigenic moiety contains glutamine at positions 149, 195, 202, or a combination thereof, as numbered according to SEQ ID NO: 179. In some embodiments, the plasmodium TRAMP polypeptide or its antigenic moiety contains glutamine at positions 149, 195, and 202, as numbered according to SEQ ID NO: 179. In some embodiments, the plasmodium TRAMP polypeptide or its antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 181.

[0169] In some embodiments, the plasmodium TRAMP polypeptide or its antigenic moiety contains asparagine at positions 112, 149, 155, 170, 195, 202, 253, 305, or combinations thereof, as numbered according to SEQ ID NO: 179. In some embodiments, the plasmodium TRAMP polypeptide or its antigenic moiety contains glutamine at positions 112, 149, 155, 170, 195, 202, 253, 305, or combinations thereof, as numbered according to SEQ ID NO: 179. In some embodiments, the plasmodium TRAMP polypeptide or its antigenic moiety contains glutamine at positions 112, 149, 155, 170, 195, 202, 253, and 305, as numbered according to SEQ ID NO: 179. In some embodiments, the plasmodium TRAMP polypeptide or its antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 180.

[0170] In some embodiments, the plasmodium TRAMP polypeptide or its antigenic moiety includes a PMX cleavage site. In some embodiments, the PMX cleavage site includes or consists of the amino acid sequence of HFLQ. In some embodiments, the plasmodium TRAMP polypeptide or its antigenic moiety includes a SUB2 cleavage site. In some embodiments, the SUB2 cleavage site includes or consists of the amino acid sequence of SEQ ID NO: 193.

[0171] In some embodiments, the linker comprises a glycine-serine linker. In some embodiments, the linker comprises or consists of the amino acid sequence given by SEQ ID NO: 138.

[0172] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 159.

[0173] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 160.

[0174] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 161.

[0175] (i) secretory signals, and (ii) polyribonucleotides encoding a polypeptide containing a plasmodium CSS polypeptide or its antigenic moiety are further provided.

[0176] In some embodiments, the polypeptide comprises (i) a secretory signal, (ii) a plasmodium CSS polypeptide or its antigenic moiety, and (iii) a transmembrane region.

[0177] In some embodiments, the polypeptide comprises (i) a secretory signal, (ii) a plasmodium CSS polypeptide or its antigenic moiety, (iii) a linker, and (iv) a transmembrane region.

[0178] In some embodiments, the secretion signal is a plasmodium secretion signal. In some embodiments, the plasmodium secretion signal is a plasmodium CSS secretion signal. In some embodiments, the plasmodium CSS secretion signal includes or consists of the amino acid sequence of SEQ ID NO: 188.

[0179] In some embodiments, the secretion signal is a viral secretion signal. In some embodiments, the secretion signal is an HSV glycoprotein D (gD) secretion signal. In some embodiments, the HSV gD secretion signal includes or consists of the amino acid sequence of SEQ ID NO: 42.

[0180] In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 182.

[0181] In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety comprises one, two, three, four, five, or six N-linked glycosylation sites. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety comprises an amino acid substitution at one or more N-linked glycosylation sites, the amino acid substitution preventing glycosylation. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety comprises a substitution of NX[T / S] to QX[T / S]. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety comprises amino acid substitutions at all N-linked glycosylation sites, the amino acid substitution preventing glycosylation.

[0182] In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains asparagine at positions 74, 88, 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains glutamine at positions 74, 88, 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains glutamine at positions 74, 88, 192, 234, 261, and 283, as numbered according to SEQ ID NO: 182. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 183.

[0183] In some embodiments, the linker comprises a glycine-serine linker. In some embodiments, the linker comprises or consists of the amino acid sequence given by SEQ ID NO: 138.

[0184] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 162.

[0185] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 163.

[0186] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 164.

[0187] In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 194.

[0188] In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 195.

[0189] In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 182.

[0190] In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety comprises one, two, three, four, five, or six N-linked glycosylation sites. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety comprises an amino acid substitution at one or more N-linked glycosylation sites, the amino acid substitution preventing glycosylation. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety comprises a substitution of NX[T / S] to QX[T / S]. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety comprises amino acid substitutions at all N-linked glycosylation sites, the amino acid substitution preventing glycosylation.

[0191] In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains asparagine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 194. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains glutamine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 194. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains glutamine at positions 74 and 88, as numbered according to SEQ ID NO: 194. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 184.

[0192] In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains asparagine at positions 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 195. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains glutamine at positions 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 195. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains glutamine at positions 192, 234, 261, and 283, as numbered according to SEQ ID NO: 195. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 185.

[0193] In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains asparagine at positions 74, 88, 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains glutamine at positions 74, 88, 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 182. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains glutamine at positions 74, 88, 192, 234, 261, and 283, as numbered according to SEQ ID NO: 182. In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains or consists of the amino acid sequence according to SEQ ID NO: 183.

[0194] In some embodiments, the first linker and / or the second linker comprises or consists of the amino acid sequence given by SEQ ID NO: 138.

[0195] In some embodiments, the transmembrane region is a heterogeneous transmembrane region. In some embodiments, the heterogeneous transmembrane region includes or consists of a viral transmembrane region. In some embodiments, the heterogeneous transmembrane region includes or consists of an HSV transmembrane region. In some embodiments, the HSV transmembrane region includes or consists of an HSV gD transmembrane region. In some embodiments, the HSV gD transmembrane region includes or consists of the amino acid sequence according to SEQ ID NO: 75.

[0196] In some embodiments, the linker comprises a glycine-serine linker. In some embodiments, the linker comprises or consists of the amino acid sequence given by SEQ ID NO: 138.

[0197] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 165.

[0198] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 166.

[0199] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 167.

[0200] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 168.

[0201] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 210.

[0202] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 211.

[0203] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence according to SEQ ID NO: 212. In some embodiments, the polypeptide comprises or consists of the amino acid sequence according to SEQ ID NO: 212.

[0204] In some embodiments, the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence according to SEQ ID NO: 213. In some embodiments, the polypeptide comprises or consists of the amino acid sequence according to SEQ ID NO: 213.

[0205] In some embodiments, the RNA construct comprises, in 5' to 3' order, (i) a 5' UTR, (ii) the polynucleotide described herein, (iii) a 3' UTR, and (iv) a polyA tail sequence.

[0206] In some embodiments, the RNA construct comprises a 5' UTR that comprises or consists of a modified human alpha-globin 5'-UTR; and a 3' UTR that comprises or consists of a first sequence derived from a split amino-terminal enhancer (AES) messenger.

[0207] In some embodiments, the RNA construct comprises a 5' UTR. In some embodiments, the 5' UTR comprises or consists of a modified human alpha-globin 5'-UTR. In some embodiments, the 5' UTR consists of the ribonucleic acid sequence according to SEQ ID NO: 111.

[0208] In some embodiments, the RNA construct comprises a 3' UTR. In some embodiments, the 3' UTR comprises or consists of a first sequence derived from a split amino-terminal enhancer (AES) messenger RNA and a second sequence derived from the mitochondrially encoded 12S ribosomal RNA. In some embodiments, the 3' UTR consists of the ribonucleic acid sequence according to SEQ ID NO: 117.

[0209] In some embodiments, the RNA construct comprises a polyA tail sequence. In some embodiments, the polyA tail sequence is a split polyA tail sequence. In some embodiments, the split polyA tail sequence consists of a ribonucleic acid sequence according to SEQ ID NO: 114.

[0210] In some embodiments, the RNA construct comprises a 5' cap.

[0211] In some embodiments, the RNA construct comprises a cap proximal sequence that includes positions +1, +2, +3, +4, and +5 of the polynucleotide.

[0212] In some embodiments, the 5' cap comprises or consists of m7(3’OMeG)(5’)ppp(5’)(2’OMeA1)pG2, wherein A1 is the +1 position of the polynucleotide and G2 is the +2 position of the polynucleotide.

[0213] In some embodiments, the cap proximal sequence comprises a sequence that includes A1 and G2 of the cap 1 structure, and A3A4U5 (SEQ ID NO: 138) at positions +3, +4, and +5, respectively, of the polynucleotide.

[0214] In some embodiments, the polynucleotides provided herein comprise modified uridine in place of all uridines. In some embodiments, the modified uridine is N1-methyl-pseudouridine, respectively.

[0215] The present disclosure also provides a composition. In some embodiments, the composition comprises one or more polynucleotides provided herein. In some embodiments, the composition comprises one or more RNA constructs provided herein.

[0216] In some embodiments, the composition comprises (i) one or more polyribonucleotides and (ii) lipid nanoparticles, polyplexes (PLX), lipid-added polyplexes (LPLX), or liposomes. In some embodiments, one or more polyribonucleotides are fully or partially encapsulated within lipid nanoparticles, polyplexes (PLX), lipid-added polyplexes (LPLX), or liposomes.

[0217] In some embodiments, the composition comprises (i) one or more polyribonucleotides and (ii) lipid nanoparticles. In some embodiments, one or more polyribonucleotides are fully or partially encapsulated within the lipid nanoparticles.

[0218] This disclosure provides pharmaceutical compositions. In some embodiments, the pharmaceutical composition comprises (i) one or more polyribonucleotides described herein, one or more RNA constructs described herein, or compositions described herein, and (ii) at least one pharmaceutically acceptable excipient.

[0219] In some embodiments, the pharmaceutical product includes a cryoprotectant. In some embodiments, the cryoprotectant is sucrose.

[0220] In some embodiments, the pharmaceutical comprises a buffered aqueous solution, which optionally comprises one or more of the following: Tris base, Tris HCl, NaCl, KCl, Na2HPO4, and KH2PO4.

[0221] The disclosure provides, in particular, a combination comprising: (i) a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more plasmodium Rh5 polypeptides or their antigenic moieties; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, and the second polypeptide comprises one or more plasmodium Rh5 intrusion complex polypeptides selected from one or more CyRPA polypeptides or their antigenic moieties, one or more Ripr polypeptides or their antigenic moieties, one or more P113 polypeptides or their antigenic moieties, one or more TRAMP polypeptides or their antigenic moieties, one or more CSS polypeptides or their antigenic moieties, or a combination thereof.

[0222] In some embodiments, the second polypeptide comprises one or more CyRPA polypeptides or their antigenic moieties. In some embodiments, the second polypeptide comprises one or more P113 polypeptides or their antigenic moieties. In some embodiments, the second polypeptide comprises one or more Ripr polypeptides or their antigenic moieties. In some embodiments, the second polypeptide comprises one or more TRAMP polypeptides or their antigenic moieties. In some embodiments, the second polypeptide comprises one or more CSS polypeptides or their antigenic moieties.

[0223] The disclosure also provides a combination comprising (i) a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more plasmodium Rh5 entry complex polypeptides or their antigenic moieties; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, and the second polypeptide comprises one or more plasmodium T cell antigens.

[0224] Furthermore, the Disclosure provides a combination comprising: (i) a first pharmaceutical composition comprising a first polyribonucleotide encoding a first polypeptide, wherein the first polypeptide comprises one or more plasmodium Rh5 intrusion complex polypeptides or their antigenic moieties; and (ii) a second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, and the second polypeptide comprises one or more plasmodium CSP polypeptides or their antigenic moieties.

[0225] In particular, the method includes administering a polyribonucleotide described herein. The disclosure also provides a method including administering an RNA construct described herein. The disclosure further provides a method including administering a composition described herein. Furthermore, a method including administering one or more doses of a pharmaceutical composition described herein.

[0226] In some embodiments, the pharmaceutical compositions provided herein are intended for use in the treatment of malaria infection, which involves administering one or more doses of the pharmaceutical composition to a target.

[0227] In some embodiments, the pharmaceutical compositions provided herein are intended for use in the prevention of malaria infection, which involves administering one or more doses of the pharmaceutical composition to a target.

[0228] In some embodiments, administering two or more doses of the pharmaceutical composition to a subject. In some embodiments, administering three or more doses of the pharmaceutical composition to a subject.

[0229] In some embodiments, the methods provided herein include administering the combinations described herein to a subject.

[0230] In some embodiments, the first pharmaceutical composition and the second pharmaceutical composition are administered on the same day. In some embodiments, the first pharmaceutical composition and the second pharmaceutical composition are administered on different days. In some embodiments, the pharmaceutical composition and the second pharmaceutical composition are administered to the subject at different locations on the subject's body.

[0231] In some embodiments, the method is a method of treating a malaria infection.

[0232] In some embodiments, the method is a method of preventing a malaria infection.

[0233] In some embodiments, the subject has a malaria infection or is at risk of developing a malaria infection.

[0234] In some embodiments, the subject is human.

[0235] In some embodiments, administration induces an anti-malaria immune response in the subject. In some embodiments, the anti-malaria immune response in the subject includes an adaptive immune response. In some embodiments, the anti-malaria immune response in the subject includes a T cell response. In some embodiments, the T cell response is or includes a CD4+ T cell response. In some embodiments, the T cell response is or includes a CD8+ T cell response. In some embodiments, the anti-malaria immune system response includes a B cell response. In some embodiments, the anti-malaria immune system response includes the production of antibodies against one or more Plasmodium antigens.

[0236] This disclosure provides the use of the pharmaceutical compositions described herein in the treatment of malaria infection.

[0237] This disclosure provides the use of the pharmaceutical compositions described herein in the prevention of malaria infection.

[0238] This disclosure provides the use of the pharmaceutical compositions described herein in inducing an antimalarial immune response in subjects.

[0239] Furthermore, this disclosure provides polypeptides encoded by the polyribonucleotides or RNA constructs described herein.

[0240] This disclosure also provides host cells comprising the polyribonucleotides or RNA constructs described herein. This disclosure further provides host cells comprising the polypeptides described herein. [Brief explanation of the drawing]

[0241] [Figure 1] This figure provides an annotated polypeptide corresponding to the Plasmodium falciparum Rh5 sequence shown in Sequence ID No. 1. Specific sequence features are annotated, including secretion signal sequences (bold red corresponds to the signal peptide described in Baum et al. 2009, with two additional red residues included in the description of the signal peptide by Wright et al. 2014), glycosylation sites (NXS / T, blue and bold, blue only for lower prediction scores, motifs in parentheses are proximal to the basidine binding site), and Rh5PMX cleavage sites (bold black). The 3D7 reference sequence often has cysteine ​​at position 203, which is tyrosine in naturally circulating parasite isolates (red), and paired cysteines are color-coded accordingly. Underlined regions are ordered and form the alpha helical composition of the protein's kite structure related to basidine binding. [Figure 2]Parts (A) to (M) are diagrams containing schematic diagrams of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino acid (aa) references contained herein refer to the Plasmodium falciparum Rh5 sequence shown in Sequence ID No. 1. (A) Includes a schematic diagram of RNA construct 1, an exemplary polypeptide encoded by polyribonucleotides as described herein. RNA construct 1 includes an HSV glycoprotein D (gD) secretion signal (e.g., Sequence ID No. 42), Rh5 N-terminal "N" disordered region (aa25-139), Rh5 ordered region (aa140-247), Rh5 linked "L" disordered region (aa248-296), and Rh5 ordered region (aa297-526). Four glycosylation sites at positions 38, 284, 214, and 297 are mutated from NX-[T / S] to NXA (where X is not proline). (B) Includes a schematic diagram of an exemplary polypeptide, RNA construct 2, encoded by the polyribonucleotide described herein. RNA construct 2 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa140-247) containing the C203Y mutation, an Rh5 ordered region (aa297-526), ​​a glycine-serine linker (e.g., SEQ ID NO: 86), and a Foldon domain. (C) Includes a schematic diagram of an exemplary polypeptide, RNA construct 3, encoded by the polyribonucleotide described herein. RNA construct 3 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa140-247) containing the C203Y mutation, and an Rh5 ordered region (297-526). (D) Includes a schematic diagram of an exemplary polypeptide, RNA construct 4, encoded by the polyribonucleotide described herein. RNA construct 4 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa140-247) containing a C203Y mutation, an Rh5 ordered region (297-526), ​​a glycine-serine linker (e.g., SEQ ID NO: 137), and a ferritin domain. (E) Includes a schematic diagram of RNA construct 5, an exemplary polypeptide encoded by the polyribonucleotides described herein.Membrane RNA construct 5 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa140-247) containing the C203Y mutation, an Rh5 ordered region (297-526), ​​a glycine-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (F) Includes a schematic diagram of an exemplary polypeptide encoded by the polyribonucleotide described herein, RNA construct 6. RNA construct 6 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa140-247) containing the C203Y mutation and the N214Q mutation, an Rh5 ordered region (aa297-526) containing the N297Q mutation, a glycine-serine linker (e.g., SEQ ID NO: 86), and a Foldon domain. (G) Includes a schematic diagram of an exemplary polypeptide encoded by the polyribonucleotide described herein, RNA construct 7. RNA construct 7 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), Rh5 ordered regions (aa140-247) containing C203Y and N214Q mutations, and Rh5 ordered regions (aa297-526) containing N297Q mutation. (H) Includes a schematic diagram of RNA construct 8, an exemplary polypeptide encoded by the polyribonucleotide described herein. Self-assembling RNA construct 8 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), Rh5 ordered regions (aa140-247) containing C203Y and N214Q mutations, Rh5 ordered regions (aa297-526) containing N297Q mutation, a glycine-serine linker (e.g., SEQ ID NO: 137), and a ferritin domain. (I) Includes a schematic diagram of RNA construct 9, an exemplary polypeptide encoded by the polyribonucleotide described herein. RNA construct 9 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), an Rh5 ordered region (aa140-247) containing the C203Y and N214Q mutations, an Rh5 ordered region (aa297-526) containing the N297Q mutation, a glycine-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (J) Includes a schematic diagram of RNA construct 10, an exemplary polypeptide encoded by the polyribonucleotides described herein.RNA construct 10 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), Rh5 ordered regions (aa140-247) containing C203Y and N214Q mutations, Rh5 ordered regions (aa297-526), ​​a glycine-serine linker (e.g., SEQ ID NO: 86), and a Foldon domain. (K) Includes a schematic diagram of RNA construct 11, an exemplary polypeptide encoded by the polyribonucleotide described herein. RNA construct 11 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), Rh5 ordered regions (aa140-247) containing C203Y and N214Q mutations, and Rh5 ordered regions (aa297-526). (L) Includes a schematic diagram of RNA construct 12, an exemplary polypeptide encoded by the polyribonucleotide described herein. RNA construct 12 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), Rh5 ordered regions (aa140-247) including C203Y and N214Q mutations, Rh5 ordered regions (aa297-526), ​​a glycine-serine linker (e.g., SEQ ID NO: 137), and a ferritin domain. (M) Includes a schematic diagram of RNA construct 13, an exemplary polypeptide encoded by the polyribonucleotides described herein. RNA construct 13 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), Rh5 ordered regions (aa140-247) including C203Y and N214Q mutations, Rh5 ordered regions (aa297-526), ​​a glycine-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). [Figure 3] This figure provides annotated polypeptides corresponding to the Plasmodium falciparum CyRPA sequence shown in Sequence ID No. 3. Specific sequence features, including secretion signal sequences (red) and glycosylation sites (NXS / T, blue), are annotated. [Figure 4]Parts (A) to (F) are diagrams containing schematic diagrams of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino acid (aa) references included refer to the Plasmodium falciparum CyRPA sequence shown in SEQ ID NO: 3. (A) Includes a schematic diagram of an exemplary polypeptide, RNA construct 14, encoded by polyribonucleotides as described herein. RNA construct 14 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa30-362), a glycine-serine linker (e.g., SEQ ID NO: 86), and a Foldon tag. (B) Includes a schematic diagram of an exemplary polypeptide, RNA construct 15, encoded by polyribonucleotides as described herein. RNA construct 15 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42) and a portion of CyRPA (aa30-362). (C) Includes a schematic diagram of an exemplary polypeptide, RNA construct 16, encoded by polyribonucleotides as described herein. RNA construct 16 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa30-362), a glycine-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (D) A schematic diagram of an exemplary polypeptide encoded by polyribonucleotides as described herein, RNA construct 17. RNA construct 17 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa30-362) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at three glycosylation sites (e.g., amino acids 145, 332, and 338 numbered according to SEQ ID NO: 3), a glycine-serine linker (e.g., SEQ ID NO: 86), and a Foldon tag. (E) A schematic diagram of an exemplary polypeptide encoded by polyribonucleotides as described herein, RNA construct 18. RNA construct 18 includes the HSV gD secretion signal (e.g., SEQ ID NO: 42) and a portion of CyRPA (aa30-362) that has mutations from NX-[T / S] to QX-[T / S] at three glycosylation sites (where X is not proline).(F) Includes a schematic diagram of an exemplary polypeptide encoded by the polyribonucleotide described herein, RNA construct 19. RNA construct 19 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa30-362) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at three glycosylation sites, a glycine-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). [Figure 5]Parts (A) to (G) are diagrams containing schematic diagrams of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino acid (aa) references contained herein refer to the Plasmodium falciparum CyRPA sequence shown in SEQ ID NO: 3. Polypeptides may optionally contain one or more linkers between various regions and / or domains. (A) Includes a schematic diagram of an exemplary polypeptide, RNA construct 22, encoded by polyribonucleotides as described herein. RNA construct 22 contains the CyRPA secretion signal (SEQ ID NO: 187) and parts of CyRPA (aa29-362). (B) Includes a schematic diagram of an exemplary polypeptide, RNA construct 23, encoded by polyribonucleotides as described herein. RNA construct 23 includes a CyRPA substructure (aa29-362) containing the CyRPA secretion signal (SEQ ID NO: 187) and three glycosylation sites (e.g., amino acids 145, 332, and 338, numbered according to SEQ ID NO: 3) from NX-[T / S] to QX-[T / S] (where X is not proline). (C) Includes a schematic diagram of an exemplary polypeptide encoded by a polyribonucleotide as described herein, RNA construct 24. RNA construct 24 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42) and three glycosylation sites (e.g., amino acids 145, 332, and 338, numbered according to SEQ ID NO: 3) from NX-[T / S] to QX-[T / S] (where X is not proline). (D) Includes a schematic diagram of an exemplary polypeptide encoded by a polyribonucleotide as described herein, RNA construct 25. RNA construct 25 comprises a full-length CyRPA (aa1-362) and an HSV gD transmembrane domain (e.g., SEQ ID NO: 75) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at three glycosylation sites (e.g., amino acids 145, 332, and 338, numbered according to SEQ ID NO: 3). (E) Includes a schematic diagram of RNA construct 26, an exemplary polypeptide encoded by the polyribonucleotides described herein.RNA construct 26 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa29-362) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at three glycosylation sites (e.g., amino acids 145, 332, and 338 numbered according to SEQ ID NO: 3), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (F) Includes a schematic diagram of an exemplary polypeptide encoded by the polyribonucleotides described herein, RNA construct 51. RNA construct 51 comprises a full-length CyRPA (aa1-362) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at three glycosylation sites (e.g., amino acids 145, 332, and 338, numbered according to SEQ ID NO: 3), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane domain (e.g., SEQ ID NO: 75). (G) Includes a schematic diagram of RNA construct 52, an exemplary polypeptide encoded by the polyribonucleotides described herein. RNA construct 52 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of CyRPA (aa29-362) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at three glycosylation sites (e.g., amino acids 145, 332, and 338 numbered according to SEQ ID NO: 3), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). [Figure 6]Parts (A) and (B) are diagrams containing schematic diagrams of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino acid (aa) references included refer to the Plasmodium falciparum P113 sequence shown in SEQ ID NO: 6. (A) Includes a schematic diagram of an exemplary polypeptide, RNA construct 20, encoded by polyribonucleotides as described herein. RNA construct 20 includes the HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of P113 (aa23-939), a glycine-serine linker (e.g., SEQ ID NO: 86), and the HSV gD transmembrane region (e.g., SEQ ID NO: 75). (B) Includes a schematic diagram of an exemplary polypeptide, RNA construct 21, encoded by polyribonucleotides as described herein. RNA construct 21 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of P113 (aa23-939) containing mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at eight glycosylation sites (e.g., amino acids 207, 268, 317, 360, 661, 697, 779, 876, and 938, numbered according to SEQ ID NO: 6), a glycine-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). [Figure 7] This figure provides annotated polypeptides corresponding to the Plasmodium falciparum Ripr sequence shown in Sequence ID No. 2. The figure annotates specific sequence features, including secretion signal sequences (red), glycosylation sites (NXS / T, blue, bold for stronger predictions), Ripr PMX cleavage sites (green), and the epidermal growth factor domain described in Chen et al. 2011, with the bolded domain being EGF7, associated with driving the strongest growth inhibition response (Healer 2019, Nagaoka 2020). [Figure 8]Parts (A) to (L) are diagrams containing schematic diagrams of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino acid (aa) references contained herein refer to the Plasmodium falciparum Ripr sequence shown in SEQ ID NO: 2. Polypeptides may optionally contain one or more linkers between various regions and / or domains. (A) Includes a schematic diagram of an exemplary polypeptide, RNA construct 27, encoded by a polyribonucleotide as described herein. RNA construct 27 contains the Ripr secretion signal (SEQ ID NO: 186) and parts of Ripr (aa20-1086), a glycine-serine linker (e.g., SEQ ID NO: 138). (B) Includes a schematic diagram of an exemplary polypeptide, RNA construct 28, encoded by a polyribonucleotide as described herein. RNA construct 28 contains the Ripr secretion signal (SEQ ID NO: 186), parts of Ripr (aa20-1086), and the HSV gD transmembrane region (e.g., SEQ ID NO: 75). (C) Includes a schematic diagram of an exemplary polypeptide, RNA construct 29, encoded by the polyribonucleotide described herein. RNA construct 29 includes a portion of Ripr (aa20~1086) with a Ripr secretion signal (SEQ ID NO: 186) and 12 glycosylation sites (e.g., amino acids 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021, numbered according to SEQ ID NO: 2) with a mutation from NX-[T / S] to QX-[T / S] (where X is not proline). (D) Includes a schematic diagram of an exemplary polypeptide, RNA construct 30, encoded by the polyribonucleotide described herein. RNA construct 30 includes the Ripr secretion signal (SEQ ID NO: 186), a portion of Ripr (aa20-1086) containing mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at 12 glycosylation sites (e.g., amino acids 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021, numbered according to SEQ ID NO: 2), and the HSV gD transmembrane domain (e.g., SEQ ID NO: 75).(E) Includes a schematic diagram of an exemplary polypeptide, RNA construct 31, encoded by the polyribonucleotide described herein. RNA construct 31 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42) and a portion of Ripr (aa20-1086). (F) Includes a schematic diagram of an exemplary polypeptide, RNA construct 32, encoded by the polyribonucleotide described herein. RNA construct 32 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa20-1086), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (G) Includes a schematic diagram of an exemplary polypeptide, RNA construct 33, encoded by the polyribonucleotide described herein. RNA construct 33 includes a portion of Ripr (aa20-1086) with an NX-[T / S] to QX-[T / S] mutation (where X is not proline) at 12 glycosylation sites (e.g., amino acids 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021, numbered according to SEQ ID NO: 2) with an HSV gD secretion signal (e.g., SEQ ID NO: 42) and 12 glycosylation sites (e.g., amino acids 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021, numbered according to SEQ ID NO: 2). (H) Includes a schematic diagram of RNA construct 34, an exemplary polypeptide encoded by the polyribonucleotides described herein. RNA construct 34 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa20-1086) containing mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at 12 glycosylation sites (e.g., amino acids 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021, numbered according to SEQ ID NO: 2), and an HSV gD transmembrane domain (e.g., SEQ ID NO: 75). (I) Includes a schematic diagram of an exemplary polypeptide encoded by the polyribonucleotides described herein, RNA construct 53. RNA construct 53 includes the Ripr secretion signal (SEQ ID NO: 186), a portion of Ripr (aa20-1086), a glycine-serine linker (e.g., SEQ ID NO: 138), and the HSV gD transmembrane region (e.g., SEQ ID NO: 75).(J) Includes a schematic diagram of an exemplary polypeptide, RNA construct 54, encoded by a polyribonucleotide as described herein. RNA construct 54 includes a Ripr secretion signal (SEQ ID NO: 186), a portion of Ripr (aa20-1086) with 12 glycosylation sites (e.g., amino acids 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021, numbered according to SEQ ID NO: 2) with an NX-[T / S] to QX-[T / S] mutation (where X is not proline), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (K) Includes a schematic diagram of an exemplary polypeptide, RNA construct 55, encoded by a polyribonucleotide as described herein. RNA construct 55 comprises the HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa20-1086), a glycine-serine linker (e.g., SEQ ID NO: 138), and the HSV gD transmembrane region (e.g., SEQ ID NO: 75). (L) Includes a schematic diagram of RNA construct 56, an exemplary polypeptide encoded by the polyribonucleotides described herein. RNA construct 56 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa20-1086) containing mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at 12 glycosylation sites (e.g., amino acids 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021, numbered according to SEQ ID NO: 2), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). [Figure 9]Parts (A) to (L) are diagrams containing schematic diagrams of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino acid (aa) references contained herein refer to the Plasmodium falciparum Ripr sequence shown in Sequence ID No. 2. Polypeptides may optionally contain one or more linkers between various regions and / or domains. (A) Includes a schematic diagram of an exemplary polypeptide, RNA construct 35, encoded by a polyribonucleotide as described herein. RNA construct 35 includes an HSV gD secretion signal (e.g., Sequence ID No. 42), a portion of Ripr (aa560-1086) with NX-[T / S] to QX-[T / S] mutations (X is not proline) at 12 glycosylation sites (e.g., amino acids 646, 964, and 1021 numbered according to Sequence ID No. 2), and an HSV gD transmembrane region (e.g., Sequence ID No. 75). (B) Includes a schematic diagram of an exemplary polypeptide, RNA construct 36, encoded by a polyribonucleotide as described herein. RNA construct 36 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of the Ripr (aa720-934), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (C) Includes a schematic diagram of an exemplary polypeptide encoded by the polyribonucleotide described herein, RNA construct 37. RNA construct 37 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of the Ripr (aa769-900), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (D) Includes a schematic diagram of an exemplary polypeptide encoded by the polyribonucleotide described herein, RNA construct 38. The membrane Ripr includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of the Ripr (aa769-856), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (E) Includes a schematic diagram of an exemplary polypeptide encoded by the polyribonucleotide described herein, membrane Ripr. RNA construct 38 includes the HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of the Ripr (aa817-900), and the HSV gD transmembrane region (e.g., SEQ ID NO: 75).(F) Includes a schematic diagram of an exemplary polypeptide, RNA construct 40, encoded by the polyribonucleotide described herein. RNA construct 40 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa817-856), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (G) Includes a schematic diagram of an exemplary polypeptide, RNA construct 57, encoded by the polyribonucleotide described herein. RNA construct 57 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa560-1086) with 12 glycosylation sites (e.g., amino acids 646, 964, and 1021 numbered according to SEQ ID NO: 2) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (H) Includes a schematic diagram of an exemplary polypeptide, RNA construct 58, encoded by the polyribonucleotide described herein. RNA construct 58 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa720-934), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (I) Includes a schematic diagram of an exemplary polypeptide, RNA construct 59, encoded by the polyribonucleotide described herein. RNA construct 59 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa769-900), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (J) Includes a schematic diagram of an exemplary polypeptide, RNA construct 60, encoded by the polyribonucleotide described herein. RNA construct 60 comprises the HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of Ripr (aa769-856), a glycine-serine linker (e.g., SEQ ID NO: 138), and the HSV gD transmembrane region (e.g., SEQ ID NO: 75). (K) Includes a schematic diagram of RNA construct 61, an exemplary polypeptide encoded by the polyribonucleotides described herein.RNA construct 61 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of the Ripr (aa817-900), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (L) Includes a schematic diagram of RNA construct 62, an exemplary polypeptide encoded by the polyribonucleotides described herein. RNA construct 62 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of the Ripr (aa817-856), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). [Figure 10] This figure provides annotated polypeptides corresponding to the Plasmodium falciparum TRAMP sequence shown in Sequence ID No. 4. Specific sequence features, including secretion signal sequences (red), glycosylation sites (NXS / T, blue), TRAMP cleavage sites (bold), and SUB2 cleavage sites (green), are annotated. [Figure 11]Parts (A) to (C) are diagrams containing schematic diagrams of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino acid (aa) references contained herein refer to the Plasmodium falciparum TRAMP sequence shown in SEQ ID NO: 4. Polypeptides may optionally contain one or more linkers between various regions and / or domains. (A) Includes a schematic diagram of an exemplary polypeptide, RNA construct 41, encoded by polyribonucleotides as described herein. RNA construct 41 contains the HSV gD secretion signal (e.g., SEQ ID NO: 42) and parts of TRAMP (aa42-352). (B) Includes a schematic diagram of an exemplary polypeptide, RNA construct 42, encoded by polyribonucleotides as described herein. RNA construct 42 includes a portion of TRAMP (aa42-352) containing an HSV gD secretion signal (e.g., SEQ ID NO: 42) and eight glycosylation sites (e.g., amino acids 112, 149, 155, 170, 195, 202, 253, and 305, numbered according to SEQ ID NO: 4) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline). (C) Includes a schematic diagram of RNA construct 43, an exemplary polypeptide encoded by the polyribonucleotides described herein. RNA construct 43 includes a portion of TRAMP (aa42-352) containing an HSV gD secretion signal (SEQ ID NO: 42) and three glycosylation sites (e.g., amino acids 149, 195, and 202, numbered according to SEQ ID NO: 4) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline). [Figure 12] This provides an annotated polypeptide corresponding to the Plasmodium falciparum CSS sequence shown in Sequence ID No. 5. Specific sequence features, including the secretion signal sequence (red), glycosylation sites (NXS / T, blue), and domains D1 and D2, are annotated, with the domains linked by purple linker regions being underlined. [Figure 13]Parts (A) to (K) are diagrams containing schematic diagrams of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino acid (aa) references contained herein refer to the Plasmodium falciparum CSS sequence shown in Sequence ID No. 5. Polypeptides may optionally contain one or more linkers between various regions and / or domains. (A) Includes a schematic diagram of an exemplary polypeptide, RNA construct 44, encoded by polyribonucleotides as described herein. RNA construct 44 contains the CSS secretion signal (Sequence ID No. 188) and parts of the CSS (aa21-290). (B) Includes a schematic diagram of an exemplary polypeptide, RNA construct 45, encoded by polyribonucleotides as described herein. RNA construct 45 includes a CSS secretion signal (SEQ ID NO: 188) and a portion of CSS (aa21-290) containing mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at six glycosylation sites (e.g., amino acids 74, 88, 192, 234, 261, and 283, numbered according to SEQ ID NO: 5). (C) Includes a schematic diagram of RNA construct 46, an exemplary polypeptide encoded by the polyribonucleotides described herein. RNA construct 46 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42) and a portion of CSS (aa21-290) containing mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at six glycosylation sites (e.g., amino acids 74, 88, 192, 234, 261, and 283, numbered according to SEQ ID NO: 5). (D) Includes a schematic diagram of an exemplary polypeptide encoded by the polyribonucleotide described herein, RNA construct 47. RNA construct 47 comprises the CSS secretion signal (SEQ ID NO: 188), a portion of CSS (aa21-290) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at six glycosylation sites (e.g., amino acids 74, 88, 192, 234, 261, and 283, numbered according to SEQ ID NO: 5), and the HSV gD transmembrane region (e.g., SEQ ID NO: 75).(E) Includes a schematic diagram of an exemplary polypeptide, RNA construct 48, encoded by a polyribonucleotide as described herein. RNA construct 48 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of CSS (aa21-290) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at six glycosylation sites (e.g., amino acids 74, 88, 192, 234, 261, and 283, numbered according to SEQ ID NO: 5), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (F) Includes a schematic diagram of an exemplary polypeptide, RNA construct 49, encoded by a polyribonucleotide as described herein. RNA construct 49 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of CSS (aa21-152) containing mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at two glycosylation sites (e.g., amino acids 74 and 88 numbered according to SEQ ID NO: 5), and an HSV gD transmembrane domain (e.g., SEQ ID NO: 75). (G) A schematic diagram of an exemplary polypeptide encoded by the polyribonucleotides described herein, RNA construct 50. RNA construct 50 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of CSS (aa153-290) containing mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at four glycosylation sites (e.g., amino acids 192, 234, 261, and 283 numbered according to SEQ ID NO: 5), and an HSV gD transmembrane domain (e.g., SEQ ID NO: 75). (H) This specification includes a schematic diagram of an exemplary polypeptide encoded by a polyribonucleotide, RNA construct 63. RNA construct 63 comprises a CSS secretion signal (SEQ ID NO: 188), a portion of CSS (aa21-290) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at six glycosylation sites (e.g., amino acids 74, 88, 192, 234, 261, and 283, numbered according to SEQ ID NO: 5), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane domain (e.g., SEQ ID NO: 75).(I) Includes a schematic diagram of an exemplary polypeptide, RNA construct 64, encoded by a polyribonucleotide as described herein. RNA construct 64 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of CSS (aa21-290) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at six glycosylation sites (e.g., amino acids 74, 88, 192, 234, 261, and 283, numbered according to SEQ ID NO: 5), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (J) Includes a schematic diagram of an exemplary polypeptide, RNA construct 65, encoded by a polyribonucleotide as described herein. RNA construct 65 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of CSS (aa21-152) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at two glycosylation sites (e.g., amino acids 74 and 88 numbered according to SEQ ID NO: 5), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (K) Includes a schematic diagram of RNA construct 66, an exemplary polypeptide encoded by the polyribonucleotides described herein. RNA construct 66 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42), a portion of CSS (aa153-290) with mutations from NX-[T / S] to QX-[T / S] (where X is not proline) at four glycosylation sites (e.g., amino acids 192, 234, 261, and 283, numbered according to SEQ ID NO: 5), a glycine-serine linker (e.g., SEQ ID NO: 138), and an HSV gD transmembrane domain (e.g., SEQ ID NO: 75). [Figure 14A]Parts (A) to (F) show the in vitro host cell viability, transfection rate, and expression of polyribonucleotide constructs encoding the plasmodium polypeptide described herein by host cells (e.g., HEK293T cells). (A) shows the percentage of viable host cells that are positive for the presence of intracellularly expressed proteins for both transfected and non-transfected (NT) cells. (B) shows the percentage of viable host cells that are positive for the presence of surface-expressed proteins for both transfected and non-transfected cells. [Figure 14C] (C) Shows the transfection rate of the indicated polyribonucleotide construct, measured by the percentage of the total host cell population that is positive for the presence of intracellularly expressed proteins. (D) Shows the transfection rate of the indicated polyribonucleotide construct, measured by the percentage of the total host cell population that is positive for the presence of surface-expressed proteins. [Figure 14E] (E) Shows total intracellular protein expression as measured by the median fluorescence intensity of the total host cell population for both transfected and untransfected cells. (F) Shows total surface protein expression as measured by the median fluorescence intensity of the total host cell population for both transfected and untransfected cells. The numbers shown on the x-axis indicate the RNA construct numbers. NT represents untransfected. [Figure 15A] Parts (A) to (D) show the transfection rates and expression of polyribonucleotide constructs encoding plasmodium polypeptides described herein by host cells (e.g., HEK293T cells). (A) Shows the transfection rate of the indicated polyribonucleotide construct, measured by the percentage of the total host cell population that is positive for the presence of intracellularly expressed proteins. (B) Shows the transfection rate of the indicated polyribonucleotide construct, measured by the percentage of the total host cell population that is positive for the presence of surface-expressed proteins. [Figure 15C](C) Shows total intracellular protein expression as measured by the median fluorescence intensity of the total host cell population for both transfected and untransfected cells. (D) Shows total surface protein expression as measured by the median fluorescence intensity of the total host cell population for both transfected and untransfected cells. The numbers shown on the x-axis indicate the RNA construct numbers. NT represents untransfected. [Figure 16A] Parts (A) and (B) show the transfection rates and expression of polyribonucleotide constructs encoding plasmodium polypeptides described herein by HEK293T cells. (A) shows the transfection rates of the shown polyribonucleotide constructs, measured by the percentage of the total host cell population that is positive for the presence of intracellularly expressed proteins. For each construct shown, cells were stained with 1:1000 dilution 35-day serum from mice immunized twice with the same construct. Cells transfected with construct 23 were stained with serum from mice immunized with construct 23. Construct 1 encoding RH5 was included as a control because ELISA and IVE data with monoclonal antibodies have already confirmed the immunogenicity of these constructs. Construct 1 was stained with both 1:2000 dilution monoclonal antibody 9AD4 and serum from mice immunized with construct 1. [Figure 16B] (B) Total protein expression is shown as measured by the median fluorescence intensity of the total host cell population for both untransfected and transfected cells. For all constructs, untransfected cells (-) are included as a negative control, and (+) indicates cells transfected with the fitted construct before staining. The numbers shown on the x-axis indicate the RNA construct number. NT represents untransfected. [Figure 17A]Parts (A) to (J) show the in vitro expression of polyribonucleotide constructs encoding the plasmodium polypeptide described herein by host cells (e.g., HEK293T cells). In vitro expression was measured by detecting cell-related proteins bound either intracellularly or on the surface, or proteins detected in the culture medium (e.g., secreted). (A) shows total cell-related protein expression determined by HiBit luminescence assay for the GARP construct, and (B) shows total secreted proteins detected in the culture medium for the GARP construct. [Figure 17C] (C) Shows total cell-associated protein expression determined by HiBit luminescence assay for the CyRPA construct, and (D) shows total secreted proteins detected in culture medium for the CyRPA construct. [Figure 17E] (E) shows the total cell-associated protein expression measured by the HiBit luminescence assay for the Ripr construct, and (F) shows the total secreted protein detected in the culture medium for the Ripr construct. [Figure 17G] (G) Shows total cell-associated protein expression determined by HiBit luminescence assay for the CSS construct, and (H) shows total secreted proteins detected in culture medium for the CSS construct. [Figure 17I] (I) shows the total cell-associated protein expression measured by HiBit luminescence assay for the TRAMP construct, and (J) shows the total secreted protein detected in culture medium for the TRAMP construct. The numbers on the x-axis indicate the RNA construct numbers. [Figure 18] This figure shows the reciprocal titer of the ELISA endpoint in serum samples from mice immunized twice with 1 μg of RH5-coding construct at day 35. Negative control samples were derived from mice immunized with a CSP-coding construct. The numbers on the x-axis indicate the RNA construct numbers. [Figure 19]Parts (A) to (D) are diagrams containing schematic diagrams of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino acid (aa) references included refer to the Plasmodium falciparum GARP sequence shown in SEQ ID NO: 23. (A) Includes a schematic diagram of an exemplary polypeptide, RNA construct 77, encoded by polyribonucleotides as described herein. RNA construct 77 includes an HSV glycoprotein D (gD) secretion signal (e.g., SEQ ID NO: 42) and a GARP-A region (aa410-673) (SEQ ID NO: 312). (B) Includes a schematic diagram of an exemplary polypeptide, RNA construct 78, encoded by polyribonucleotides as described herein. RNA construct 78 includes an HSV gD secretion signal (e.g., SEQ ID NO: 42) and a GARP-A region (aa410-673) (SEQ ID NO: 312) containing the N504Q mutation (SEQ ID NO: 313). (C) Includes a schematic diagram of an exemplary polypeptide, RNA construct 82, encoded by polyribonucleotides as described herein. RNA construct 82 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a GARP-A region (aa410~673) (SEQ ID NO: 312) containing the N504Q mutation (SEQ ID NO: 312), a glycine-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). (D) A schematic diagram of an exemplary polypeptide encoded by the polyribonucleotides described herein, RNA construct 84. RNA construct 84 comprises an HSV gD secretion signal (e.g., SEQ ID NO: 42), a GARP-A region (aa52~673) (SEQ ID NO: 342) containing the N183Q, N233Q, N242Q, and N504Q mutations (SEQ ID NO: 309), as well as a glycine-serine linker (e.g., SEQ ID NO: 86), and an HSV gD transmembrane region (e.g., SEQ ID NO: 75). [Figure 20]Parts (A) to (C) are figures containing schematic diagrams of exemplary polypeptides encoded by polyribonucleotides provided herein. All amino acid (aa) references included refer to the Plasmodium falciparum TRAMP (e.g., PTRAMP) sequence shown in SEQ ID NO: 4. (A) Includes a schematic diagram of an exemplary polypeptide, RNA construct 133, encoded by polyribonucleotides as described herein. RNA construct 133 includes an HSV glycoprotein D (gD) secretion signal (e.g., SEQ ID NO: 42) and a TRAMP region (aa42-352). (B) Includes a schematic diagram of an exemplary polypeptide, RNA construct 134, encoded by polyribonucleotides as described herein. RNA construct 134 includes an HSV glycoprotein D (gD) secretion signal (e.g., SEQ ID NO: 42), as well as TRAMP regions (aa42-352) containing N112Q, N129Q, N155Q, N170Q, N195Q, N202Q, N253Q, and N305Q mutations. (C) Includes a schematic diagram of RNA construct 135, an exemplary polypeptide encoded by the polyribonucleotides described herein. RNA construct 135 includes an HSV glycoprotein D (gD) secretion signal (e.g., SEQ ID NO: 42), as well as TRAMP regions (aa42-352) containing N149Q, N195Q, and N202Q mutations. [Modes for carrying out the invention]

[0242] definition The compounds disclosed herein include those generally described above, and are further illustrated by the classes, subclasses, and species disclosed herein. Where used herein, unless otherwise specified, the following definitions apply. For the purposes of this disclosure, chemical elements are identified according to the Periodic Table, CAS versions, and Handbook of Chemistry and Physics, 75th edition. Furthermore, the general principles of organic chemistry are found in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry," 5th edition, edited by Smith, MB and March, J., John Wiley & Sons, New York: 2001, the entire contents of each of these are incorporated herein by reference.

[0243] Unless otherwise specified, the structures shown herein are intended 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 this disclosure. Thus, single stereochemical isomers of the compounds provided, as well as enantiomers, diastereomers, and geometric (or conformational) mixtures, are within the scope of this disclosure. For example, in some cases, the compounds provided represent one or more stereoisomers of the compound, and unless otherwise specified, each stereoisomer is represented individually and / or as a mixture. Unless otherwise specified, all tautomeric forms of the compounds provided are within the scope of this disclosure.

[0244] Unless otherwise indicated, the structures shown herein are intended to represent compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the structure including the substitution of hydrogen with deuterium or tritium, or the substitution of carbon with 13C or 14C enriched carbon, are within the scope of this disclosure.

[0245] Approximately: When used herein in relation to a value, the term "approximately" refers to a value similar to the value being referenced in the context of the referenced value. In general, a person skilled in the art familiar with the context will understand the relevant degree of dispersion encompassed by "approximately" in that context. For example, in some embodiments, the term "approximately" may encompass a range of values ​​within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less than 1% of the value mentioned.

[0246] Drugs: As used herein, the term “drug” may refer to a physical entity. In some embodiments, a drug may be characterized by certain features and / or effects. For example, as used herein, the term “therapeutic agent” refers to a physical entity that has a therapeutic effect and / or induces a desired biological and / or pharmacological effect. In some embodiments, a drug may be a compound, molecule, or entity of any chemical class, including, for example, small molecules, polypeptides, nucleic acids, sugars, lipids, metals, or combinations or complexes thereof.

[0247] Amino Acids: In its broadest sense, as used herein, the term “amino acid” refers to compounds and / or substances that can, are incorporated into, or have been incorporated into a polypeptide chain, for example, through the formation of one or more peptide bonds. In some embodiments, amino acids have a general structure H2N-C(H)(R)-COOH. In some embodiments, amino acids are naturally occurring amino acids. In some embodiments, amino acids are non-natural amino acids; in some embodiments, amino acids are D-amino acids; in some embodiments, amino acids are L-amino acids. “Standard amino acids” refers to any of the 20 standard L-amino acids commonly found in naturally occurring peptides. “Non-standard amino acids” refers to any amino acid other than standard amino acids, whether synthetically prepared or obtained from natural sources. In some embodiments, amino acids in a polypeptide, including carboxyl-terminal and / or amino-terminal amino acids, may undergo structural modifications compared to the general structure described above. For example, in some embodiments, amino acids may be modified compared to the general structure by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and / or substitution (e.g., amino groups, carboxylic acid groups, one or more protons, and / or hydroxyl groups). In some embodiments, such modifications may alter the cyclic half-life of the modified amino acid-containing polypeptide compared to, for example, a polypeptide containing otherwise identical unmodified amino acids. In some embodiments, such modifications do not significantly alter the relevant activity of the modified amino acid-containing polypeptide compared to a polypeptide containing otherwise identical unmodified amino acids. As is evident from the context, in some embodiments, the term “amino acid” may be used to refer to free amino acids; in some embodiments, the term “amino acid” may be used to refer to amino acid residues of a polypeptide.

[0248] Antigen: As used herein, the term “antigen” means (i) a drug that induces an immune response, and / or (ii) a drug that binds to a T cell receptor (for example, when presented by an MHC molecule) or an antibody.

[0249] Antimalarial immune response: As used herein, the term “antimalarial immune response” refers to an immune response to one or more antigens derived from Plasmodium.

[0250] Related: Two events or entities are “related” to each other if, as the term is used herein, the existence, level, degree, type, and / or form of one correlates with that of the other. For example, a particular entity (e.g., polypeptide, gene signature, metabolite, microorganism, etc.) is considered related to a particular disease, disorder, or condition if its existence, level, and / or form correlates with the incidence, susceptibility, severity, stage, etc. of a disease, disorder, or condition (e.g., across a relevant population). In some embodiments, two or more entities are physically “related” to each other if they interact directly or indirectly such that they are and / or remain physically close to each other. In some embodiments, two or more entities that are physically related to each other are covalently bonded to each other; in some embodiments, two or more entities that are physically related to each other are not covalently bonded to each other but are non-covalently related by, for example, hydrogen bonding, van der Waals interactions, hydrophobic interactions, magnetism, and combinations thereof.

[0251] Characteristic parts: As used herein, the term “characteristic parts” means, in its broadest sense, a portion of a polypeptide or region whose presence (or absence) correlates with the presence (or absence) of a particular feature, attribute, or activity of the polypeptide or region. In some embodiments, a characteristic part of a polypeptide or region is a portion found in a polypeptide or region and related polypeptides or regions that share a particular feature, attribute, or activity, but not in those that do not share that particular feature, attribute, or activity. In certain embodiments, a characteristic part shares at least one functional feature with the intact polypeptide or region. For example, in some embodiments, a “characteristic part” of a polypeptide or region contains a sequence of amino acid stretches, or a collection of amino acid stretches, that together are characteristic of the polypeptide or region. In some embodiments, each such sequence of stretches generally contains at least 2, 5, 10, 15, 20, 50 or more amino acids. Generally, a characteristic part of a polypeptide or region shares at least one functional feature with the related intact polypeptide or region, in addition to the sequence and / or structural identity described above. In some embodiments, a characteristic part may be biologically active. In some embodiments, the fragments described herein may be parts. Thus, in some embodiments, a characteristic fragment may be a "characteristic part."

[0252] Combination Therapy: As used herein, the term “combination therapy” refers to a situation in which a subject is simultaneously exposed to two or more treatment regimens (e.g., two or more therapeutic agents (e.g., two or more antibody drugs)). In some embodiments, two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all “dose” of the first regimen are administered before any dose of the second regimen is administered); in some embodiments, such drugs are administered in overlapping dosing regimens. In some embodiments, the administration of combination therapy may involve the administration of one or more drugs or modalities to a subject that is administered in combination with one or more other drugs or modalities. For clarity, combination therapy does not require that individual drugs be administered together (or necessarily simultaneously) in a single composition, but in some embodiments, two or more drugs or their active parts may be administered together in a combination composition.

[0253] Comparable: As used herein, the term “comparable” means two or more sets of agents, entities, situations, conditions, etc., that may not be identical to one another but are similar enough to allow for comparison between them, and as a result, a person skilled in the art will understand that conclusions can be reasonably drawn based on observed differences or similarities. In some embodiments, a comparable set of conditions, situations, individuals or groups is characterized by several substantially identical features and one or a few varying features. A person skilled in the art will understand, in context, what degree of identity is required in any given situation for two or more sets of such agents, entities, situations, conditions, etc., to be considered comparable. For example, a person skilled in the art will understand that sets of situations, individuals or groups are comparable if they are characterized by a sufficient number and variety of substantially identical features to ensure that differences in results or observed phenomena obtained under or with respect to different sets of situations, individuals or groups are caused by or indicate variations in these varying features.

[0254] Corresponding: As used herein, the term “corresponding” refers to a relationship between two or more entities. For example, the term “corresponding” may be used to specify the position / identity of a structural element in one compound or composition to another compound or composition (e.g., a suitable reference compound or composition). For example, in some embodiments, a monomer 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 a suitable reference polymer. For example, those skilled in the art will understand that, for simplification, residues in polypeptides are often specified using a standard numbering system based on reference-related polypeptides, and as a result, for example, an amino acid “corresponding” to the residue at position 190 does not actually need to be the 190th amino acid in a particular amino acid chain, but rather corresponds to the residue found at position 190 in the reference polypeptide; those skilled in the art will readily understand how to identify “corresponding” amino acids. For example, a person skilled in the art will recognize a variety of sequence alignment strategies, including software programs such as 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, which can be used, for example, to identify “corresponding” residues in polypeptides and / or nucleic acids in accordance with this disclosure. A person skilled in the art will also understand that in some cases the term “corresponding” may be used to describe an event or entity that shares a similarity with another event or entity (e.g., a suitable reference event or entity).For example, a gene or protein in one organism may be described as "corresponding" to a gene or protein from another organism in some embodiments to indicate that it performs a similar role or function, and / or exhibits a certain degree of sequence identity or homology, or shares certain characteristic sequence elements.

[0255] Dosage regimen: Those skilled in the art will understand that the term “dosage regimen” (or “treatment regimen”) may be used to refer to a set of individual unit doses (typically two or more) administered to a subject, typically divided by time. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses.

[0256] Code: As used herein, the terms “code” or “to code” refer to sequence information of a first molecule that leads to the production of a second molecule having a defined sequence of nucleotides (e.g., polyribonucleotides) or a defined sequence of amino acids. For example, a DNA molecule can code for an RNA molecule (e.g., by a transcription process involving the DNA-dependent RNA polymerase enzyme). An RNA molecule can code for a polypeptide (e.g., by a translation process). Thus, a gene, cDNA, or RNA molecule codes for a polypeptide if the transcription and translation of the RNA corresponding to that gene produces a polypeptide in a cell or other biological system. In some embodiments, the coding region of a polyribonucleotide coding for a target antigen refers to a coding strand whose nucleotide sequence is identical to the polyribonucleotide sequence of such a target antigen. In some embodiments, the coding region of a polyribonucleotide coding for a target antigen refers to a non-coding strand of such a target antigen that can be used as a template for the transcription of the gene or cDNA.

[0257] Expression: As used herein, the term “expression” of a nucleic acid sequence refers to the production of a gene product from a nucleic acid sequence. In some embodiments, the gene product may be a transcript, e.g., a polyribonucleotide provided herein. In some embodiments, the gene product may be a polypeptide. In some embodiments, the expression of a nucleic acid sequence involves one or more of the following: (1) production of an RNA template from the DNA sequence (e.g., by transcription); (2) processing of the RNA transcript (e.g., by splicing, editing, etc.); (3) translation of the RNA into a polypeptide or protein; and / or (4) post-translational modification of the polypeptide or protein.

[0258] Heterogeneous: As used herein, the term “heterogeneous” refers to secretory signals or transmembrane regions from organisms other than viruses or plasmodiums.

[0259] Homology: As used herein, the term “homology” or “homologous” refers to the overall relationship between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules. In some embodiments, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are considered “homologous” to one another if their sequences are identical by at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99%. In some embodiments, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are considered “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., including residues having chemical properties associated with their corresponding positions). For example, as is well known to those skilled 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 “nonpolar” side chains. The substitution of one amino acid for another amino acid of the same type can often be considered a “homologous” substitution.

[0260] Identity: As used herein, the term “identity” refers to the overall relationship between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules. In some embodiments, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are considered “substantially identical” to one another if their sequences are identical by at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%. The percentage of identity of two nucleic acid or polypeptide sequences can be calculated, for example, by aligning the two sequences for the purpose of best comparison (for example, gaps can be introduced in one or both of the first and second sequences for best alignment, and non-identical sequences can be ignored for the purpose of comparison). In certain embodiments, the length of the sequences to be aligned for comparison purposes 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 the reference sequence. The nucleotides at the corresponding positions are then compared. The molecules are identical at a position if the position in the first sequence is occupied by the same residue (e.g., a nucleotide or amino acid) as the corresponding position in the second sequence. The percentage of identity between 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 that must be introduced for optimal alignment of the two sequences. The comparison of sequences and the determination of the percentage of identity between two sequences can be achieved using mathematical algorithms. For example, the percentage of identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller, 1989, which is incorporated into the ALIGN program (version 2.0).In some exemplary embodiments, nucleic acid sequence comparison performed using the ALIGN program utilizes a PAM120 weighted residue table, a 12-gap length penalty, and a 4-gap penalty. Alternatively, the identity percentage between two nucleotide sequences can be determined using the GAP program of the GCG software package, which uses the NWSgapdna.CMP matrix.

[0261] Increase, Induction, or Decrease: As used herein, these terms or grammatically comparable comparative terms indicate a value compared to a comparable reference measure. For example, in some embodiments, the evaluation value achieved with a provided composition (e.g., a pharmaceutical composition) may be an “increase” compared to an evaluation value obtained with a comparable reference composition. Or, or further, in some embodiments, the evaluation value achieved in a subject may be an “increase” compared to an evaluation value obtained in the same subject under different conditions (e.g., before or after an event; or in the presence or absence of an event such as administration of the composition described herein (e.g., a pharmaceutical composition)) or compared to an evaluation value obtained in a different comparable subject (e.g., in a comparable subject different from the subject in question in the absence of prior exposure to a state, e.g., the absence of administration of the composition described herein (e.g., a pharmaceutical composition)). In some embodiments, comparative terms refer to a statistically significant difference (e.g., one with sufficient frequency and / or magnitude to achieve statistical significance). A person skilled in the art will be able to recognize or readily determine, in a given context, the degree and / or frequency of the difference necessary or sufficient to achieve such statistical significance. In some embodiments, the term “decrease” or equivalent refers to a decrease in the level of evaluation value of at least 5%, at least 10%, at least 20%, at least 50%, at least 75%, or more, compared to a comparable reference. In some embodiments, the term “decrease” or equivalent refers to a complete or essentially complete inhibition, i.e., a decrease to zero or a decrease to essentially zero. In some embodiments, the term “increase” or “induction” refers to an increase in the level of evaluation value of 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 more, compared to a comparable reference.

[0262] In order: As used herein with respect to polynucleotides or polyribonucleotides, “in order” refers to the order of features along the polynucleotide or polyribonucleotide from 5' to 3'. As used herein with respect to polypeptides, “in order” refers to the order of features moving along the polypeptide from the most N-terminal feature to the most C-terminal feature. “In order” does not mean that there may be no further features among the listed features. For example, if polynucleotide features A, B, and C are described herein as “in order, feature A, feature B, and feature C,” this description does not preclude, for example, feature D from being located between feature A and feature B.

[0263] Isolated: The term “isolated” means modified or removed from its natural state. For example, nucleic acids or peptides that are naturally present in a living animal are not “isolated,” but the same nucleic acids or peptides that have been partially or completely separated from their natural coexisting substances are “isolated.” Isolated nucleic acids or proteins may exist in a substantially purified form or in a non-natural environment, such as a host cell.

[0264] Linker: As used herein, the term "linker" refers to a portion of a polypeptide that connects different regions, parts, or antigens to each other.

[0265] Lipids: As used herein, the terms “lipids” and “lipid-like substances” are broadly defined as molecules that also contain one or more hydrophobic moieties or hydrophobic groups, and optionally one or more hydrophilic moieties or hydrophilic groups. Molecules containing both hydrophobic and hydrophilic moieties are also typically referred to as amphiphilic substances.

[0266] Merozoite-specific plasmodium antigen: As used herein, the term “merozoite-specific plasmodium antigen” refers to an antigen expressed during the merozoite stage of the plasmodium life cycle. In some embodiments, the merozoite-specific plasmodium antigen is the Rh5 entry complex antigen.

[0267] Multimerization region: As used herein, the term “multimerization region” refers to a region that leads the assembly of multimers into a complex, where each multimer contains a polypeptide associated with the multimerization region.

[0268] RNA lipid nanoparticles: As used herein, the term “RNA lipid nanoparticles” refers to nanoparticles comprising at least one lipid and one or more RNA molecules, e.g., one or more polyribonucleotides provided herein. In some embodiments, RNA lipid nanoparticles comprise at least one cationic aminolipid. In some embodiments, RNA lipid nanoparticles comprise at least one cationic aminolipid, at least one helper lipid, and at least one polymer conjugate lipid (e.g., PEG conjugate lipid). In various embodiments, RNA lipid nanoparticles described herein may 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 this disclosure, RNA lipid nanoparticles may have particle sizes (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 nm to about 90 nm, about 80 nm to about 90 nm, or about 70 nm to about 80 nm. In some embodiments, the average size of the lipid nanoparticles is determined by measuring the average particle size. In some embodiments, RNA lipid nanoparticles may be prepared by mixing lipids with RNA molecules as described herein.

[0269] Neutralization: As used herein, the term “neutralization” refers to the event in which a binder, such as an antibody, binds to a biologically active site of a parasite, such as a receptor-binding protein, thereby inhibiting parasitic infection of the cell. In some embodiments, the term “neutralization” refers to the event in which the binder eliminates or significantly reduces the cell’s ability to infect.

[0270] Nucleic acid / polynucleotide: As used herein, the term “nucleic acid” refers to a polymer of at least 10 nucleotides or more. In some embodiments, the nucleic acid is or contains DNA. In some embodiments, the nucleic acid is or contains RNA. In some embodiments, the nucleic acid is or contains peptide nucleic acid (PNA). In some embodiments, the nucleic acid is or contains single-stranded nucleic acid. In some embodiments, the nucleic acid is or contains double-stranded nucleic acid. In some embodiments, the nucleic acid contains both single-stranded and double-stranded portions. In some embodiments, the nucleic acid contains a backbone comprising one or more phosphodiester bonds. In some embodiments, the nucleic acid contains a backbone comprising both phosphodiester bonds and non-phosphodiester bonds. For example, in some embodiments, the nucleic acid may contain a backbone comprising one or more phosphorothioate bonds or 5'-N-phosphoramidite bonds and / or one or more peptide bonds, as in “peptide nucleic acid,” for example. In some embodiments, the nucleic acid comprises one or more, or all, of the native residues (e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uracil). In some embodiments, the nucleic acid comprises one or more, or all, of the non-native residues. In some embodiments, non-natural residues include nucleoside analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolopyrimidine, 3-methyladenosine, 5-methylcytidine, C-5 propynylcytidine, C-5 propynyluridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyluridine, C5-propynylcytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 6-O-methylguanine, 2-thiocytidine, methylated bases, inserted bases, and combinations thereof). In some embodiments, the non-natural residues include one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) compared to those in the natural residues.In some embodiments, the nucleic acid has a nucleotide sequence encoding a functional gene product such as RNA or polypeptide. In some embodiments, the nucleic acid has a nucleotide sequence containing one or more introns. In some embodiments, the nucleic acid may be prepared by isolation from a natural source, enzymatic synthesis (e.g., by polymerization based on a complementary template in vivo or in vitro), growth in recombinant cells or recombinant systems, or chemosynthesis. In some embodiments, nucleic acids are 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, 400 The residue or nucleotide length is 0, 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.

[0271] Pharmaceutically Effective Dose: The terms “pharmaceutically effective dose” or “therapeutic dose” refer to the amount that, alone or in combination with additional doses, achieves the desired response or effect. In the case of the treatment of a particular disease (e.g., malaria), the desired response, in some embodiments, relates to the inhibition of the course of the disease (e.g., malaria). In some embodiments, such inhibition may include slowing the progression of the disease (e.g., malaria) and / or interrupting or reversing the progression of the disease (e.g., malaria). In some embodiments, the desired response in the treatment of a disease (e.g., malaria) may be, or include, the delay or prevention of the onset of the disease (e.g., malaria) or condition (e.g., malaria-related condition). The effective dose of the compositions described herein (e.g., pharmaceutical compositions) depends, for example, the disease (e.g., malaria) or condition (e.g., malaria-related condition) being treated, the severity of such disease (e.g., malaria) or condition (e.g., malaria-related condition), the individual parameters of the patient, such as age, physiological status, size and weight, duration of treatment, type of concomitant treatment (if any), specific route of administration, and similar factors. Therefore, the dose of the compositions described herein (e.g., pharmaceutical compositions) may depend on a variety of such parameters. If the response in the patient is insufficient with the initial dose, a higher dose (or a substantially higher dose achieved by a different, more localized route of administration) may be used.

[0272] Polypeptide: As used herein, the term "polypeptide" refers to a polymer chain of amino acids. In some embodiments, the polypeptide has a naturally occurring amino acid sequence. In some embodiments, the polypeptide has an amino acid sequence that is not naturally occurring. In some embodiments, the polypeptide has an amino acid sequence that is engineered in that it is designed and / or produced through human action. In some embodiments, the polypeptide may contain or consist of natural amino acids, non-natural amino acids, or both. In some embodiments, the polypeptide may contain or consist of only natural amino acids or only non-natural amino acids. In some embodiments, the polypeptide may contain D-amino acids, L-amino acids, or both. In some embodiments, the polypeptide may contain only D-amino acids. In some embodiments, the polypeptide may contain only L-amino acids. In some embodiments, the polypeptide may contain one or more pendant groups or other modifications, e.g., modifications that modify or are attached to one or more amino acid side chains, at the N-terminus of the polypeptide, the C-terminus of the polypeptide, or any combination thereof. In some embodiments, such pendant groups or modifications include acetylation, amidation, lipidation, methylation, pegylation, and combinations thereof. In some embodiments, the polypeptide may be cyclic and / or contain a cyclic portion. In some embodiments, the polypeptide is not cyclic and / or does not contain a cyclic portion. In some embodiments, the polypeptide is linear. In some embodiments, the polypeptide may be or contain a staple polypeptide. In some embodiments, the term “polypeptide” may be affixed to the name of a reference polypeptide, activity, or structure; in such cases, the term is used herein to refer to polypeptides that share a related activity or structure and can therefore be considered members of the same class or family of polypeptides.For each such class, this specification provides exemplary polypeptides within the class whose amino acid sequence and / or function are known, and / or which will be recognizable to those skilled in the art; in some embodiments, such exemplary polypeptides are reference polypeptides for the polypeptide class or polypeptide family. In some embodiments, members of the polypeptide class or polypeptide family exhibit significant sequence homology or identity with the reference polypeptide of that class, and in some embodiments with all polypeptides within that class, or share common sequence motifs (e.g., characteristic sequence elements) and / or common activity (in some embodiments, at a comparable level or within a specified range). For example, in some embodiments, the member polypeptide includes at least about 30–40%, often exceeding about 50%, and exhibiting a degree of overall sequence homology or sequence identity with the reference polypeptide of 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, and / or including at least one region (for example, a conserved region that may be or may contain a characteristic sequence element in some embodiments) exhibiting very high sequence identity, often exceeding 90%, or even exceeding 95%, 96%, 97%, 98%, or 99%. Such a conserved region typically comprises at least 3 to 4 amino acids, often up to 35 or more; in some embodiments, the conserved region comprises at least one stretch of 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 consecutive amino acids. In some embodiments, the related polypeptide may comprise or consist of fragments of the parent polypeptide. In some embodiments, the polypeptide is a plasmodium polypeptide construct as described herein.A plasmodium polypeptide construct is a polypeptide comprising one or more malaria proteins or one or more portions thereof. In some embodiments, the plasmodium polypeptide construct described herein comprises at least one region or antigenic portion of a plasmodium Rh5 entry complex polypeptide. In some embodiments, the plasmodium polypeptide construct further comprises one or more further 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.

[0273] Preventing: As used herein, the terms “preventing” or “prevention,” when used in relation to the development of a disease, disorder and / or condition, mean reducing the risk of developing a disease, disorder and / or condition, and / or delaying the development of one or more characteristics or symptoms of the disease, disorder or condition. Prevention may be considered complete if the development of the disease, disorder or condition is delayed for a given period of time. In some embodiments, prevention means reducing the risk of developing clinical malaria.

[0274] Reference: As used herein, the term “reference” refers to a standard or control on which a comparison is made. For example, in some embodiments, a drug, animal, individual, population, sample, sequence, or value of interest is compared to a reference or control drug, animal, individual, population, sample, sequence, or value. In some embodiments, the reference or control is tested and / or determined substantially concurrently with the test or determination of interest. In some embodiments, the reference or control is a past reference or control optionally embodied in a tangible medium. Typically, as will be understood by those skilled in the art, the reference or control is determined or characterized under conditions or circumstances comparable to those being evaluated. Those skilled in the art will understand that there is sufficient similarity to justify reliance on and / or comparison to a particular possible reference or control.

[0275] Rh5 entry complex: As used herein, this refers to a complex present in a plasmodium merozoite containing Rh5. In some embodiments, the Rh5 entry complex facilitates merozoite binding to and / or entry of erythrocytes. Polypeptides in the Rh5 entry complex may include, but are not limited to, one or more of the following: plasmodium reticulocyte-binding protein homolog 5 (Rh5), plasmodium cysteine-rich protective antigen (CyRPA), plasmodium Rh5 interacting protein (Ripr), plasmodium P113, plasmodium thrombospondin-associated apical merozoite protein (TRAMP), and plasmodium cysteine-rich microsecretory protein (CSS). Exemplary Rh5 entry complexes include the RCR complex and the PCRCR complex. The RCR complex is a trimer complex containing Rh5, cysteine-rich protective antigen (CyRPA), and Rh5 interacting protein (Ripr). The PCRCR complex is a pentameric complex containing Rh5, cysteine-rich microsecretion (CSS), Ripr, CyRPA, and plasmodium thrombospondin-associated apical merozoite protein (PTRAMP).

[0276] Ribonucleic acid (RNA) or polyribonucleotide: As used herein, the terms “ribonucleic acid,” “RNA,” or “polyribonucleotide” refer to polymers of ribonucleotides. In some embodiments, RNA is single-stranded. In some embodiments, RNA is double-stranded. In some embodiments, RNA includes both single-stranded and double-stranded portions. In some embodiments, RNA may include the skeletal structures described above in the definition of “nucleic acid / polynucleotide.” RNA may be regulatory RNA (e.g., siRNA, microRNA, etc.) or messenger RNA (mRNA). In some embodiments, RNA is mRNA. In some embodiments, if RNA is mRNA, RNA typically includes a poly(A) region at its 3' end. In some embodiments, if RNA is mRNA, RNA typically includes a cap structure recognized in the art at its 5' end for recognizing mRNA and binding to a ribosome, for example, to initiate translation. In some embodiments, RNA is synthetic RNA. Examples of synthetic RNA include RNA synthesized in vitro (e.g., by enzymatic synthesis and / or chemical synthesis). In some embodiments, the polyribonucleotide encodes a polypeptide, which is preferably a plasmodium polypeptide construct.

[0277] Ribonucleotides: As used herein, the term “ribonucleotide” encompasses both unmodified and modified ribonucleotides. Examples of unmodified ribonucleotides include the purine bases adenine (A) and guanine (G), and the pyrimidine bases cytosine (C) and uracil (U). Modified ribonucleotides may include, for example, one or more modifications, including (a) terminal modifications, e.g., 5' terminal modifications (e.g., phosphorylation, dephosphorylation, conjugation, reverse bond), 3' terminal modifications (e.g., conjugation, reverse bond), (b) base modifications, e.g., substitution with a modified base, stabilizing base, destabilizing base, or base that forms a base pair with the extended repertoire of a partner, or a conjugate base, (c) sugar modifications (e.g., at the 2' or 4' position) or sugar substitution, and (d) nucleoside bond modifications, including modification or substitution of phosphodiester bonds. The term "ribonucleotide" also encompasses ribonucleotide triphosphates, including modified and unmodified ribonucleotide triphosphates.

[0278] Secretory signal: As used herein, the term “secretory signal” refers to an amino acid sequence motif that targets the relevant polypeptide for translocation to the secretory pathway.

[0279] Subject: As used herein, the term “subject” refers to an organism to which the composition described herein is administered, for example, for experimental, diagnostic, preventive, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and domesticated pets) and humans. In preferred embodiments, the subject is a human subject. In some embodiments, the subject suffers from a disease, disorder, or condition (e.g., malaria and / or malaria-related conditions). In some embodiments, the subject is susceptible to a disease, disorder, or condition (e.g., malaria and / or malaria-related conditions). In some embodiments, the subject exhibits one or more symptoms or characteristics of a disease, disorder, or condition (e.g., malaria and / or malaria-related conditions). In some embodiments, the subject exhibits one or more nonspecific symptoms of a disease, disorder, or condition (e.g., malaria and / or malaria-related conditions). In some embodiments, the subject exhibits no symptoms or characteristics of a disease, disorder, or condition (e.g., malaria and / or malaria-related conditions). In some embodiments, the subject is a person having one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition (e.g., malaria and / or malaria-related conditions). In some embodiments, the subject is a patient. In some embodiments, the subject is an individual who is being diagnosed with and / or receiving treatment, and / or has previously received treatment.

[0280] Affected: An individual who is “affected” by a disease, disorder and / or condition (e.g., malaria and / or malaria-related condition) is diagnosed with the disease, disorder and / or condition and / or exhibits one or more symptoms of the disease, disorder and / or condition.

[0281] Susceptible to: Individuals who are “suspended” to a disease, disorder, and / or condition (e.g., malaria and / or malaria-related conditions) are individuals who are at a higher risk of developing a disease, disorder, and / or condition (e.g., malaria and / or malaria-related conditions) than the general population. In some embodiments, individuals susceptible to a disease, disorder, and / or condition (e.g., malaria and / or malaria-related conditions) may not be diagnosed with the disease, disorder, and / or condition (e.g., malaria and / or malaria-related conditions). In some embodiments, individuals susceptible to a disease, disorder, and / or condition (e.g., malaria and / or malaria-related conditions) may exhibit symptoms of the disease, disorder, and / or condition (e.g., malaria and / or malaria-related conditions). In some embodiments, individuals susceptible to a disease, disorder, and / or condition (e.g., malaria and / or malaria-related conditions) may not exhibit symptoms of the disease, disorder, and / or condition (e.g., malaria and / or malaria-related symptoms). In some embodiments, individuals susceptible to a disease, disorder, and / or condition (e.g., malaria and / or malaria-related conditions) will develop the disease, disorder, and / or condition (e.g., malaria and / or malaria-related conditions). In some embodiments, individuals susceptible to a disease, disorder, and / or condition (e.g., malaria and / or malaria-related conditions) will not develop the disease, disorder, and / or condition (e.g., malaria and / or malaria-related conditions).

[0282] Treatment: The term “treatment” refers to the administration or delivery of a drug or intervention that has a therapeutic effect and / or induces a desired biological and / or pharmacological effect (for example, one that has been statistically demonstrated to have such an effect when administered to a relevant population). In some embodiments, a therapeutic agent or treatment is any substance that can be used to alleviate, improve, reduce, inhibit, prevent, delay the onset, reduce the severity, and / or decrease the incidence of one or more symptoms or characteristics of a disease, disorder and / or condition (e.g., malaria and / or malaria-related conditions). In some embodiments, a therapeutic agent or treatment is a medical intervention that can be implemented to alleviate, reduce, inhibit, present, delay the onset, reduce the severity, and / or decrease the incidence of one or more symptoms or characteristics of a disease, disorder and / or condition.

[0283] Transmembrane region: As used herein, the term “transmembrane region” refers to a region of polypeptide that spans a biological membrane, such as the plasma membrane of a cell.

[0284] To treat: As used herein, the terms “to treat,” “to treat,” or “to treat” refer to any method used to partially or completely alleviate, improve, reduce, inhibit, prevent, delay the onset, reduce the severity, and / or reduce the incidence of one or more symptoms or features of a disease, disorder and / or condition (e.g., malaria and / or malaria-related conditions). Treatment may be administered to subjects who do not show signs of a disease, disorder and / or condition (e.g., malaria and / or malaria-related conditions). In some embodiments, treatment may be administered to subjects who show only the initial signs of a disease, disorder and / or condition (e.g., malaria and / or malaria-related conditions), for example, to reduce the risk of developing a disease, disorder and / or condition associated with the disease, disorder and / or condition. In some embodiments, treatment may be administered to subjects in the later stages of a disease, disorder and / or condition (e.g., malaria and / or malaria-related conditions).

[0285] Mutant: As used herein, the term “mutant” refers to a molecule that exhibits significant structural (e.g., primary or secondary) identity with a reference molecule but is structurally different from the reference molecule. For example, a mutant polypeptide or nucleic acid may differ from a reference polypeptide or nucleic acid as a result of one or more differences in the amino acid sequence or nucleotide sequence, and / or one or more differences in the chemical moieties (e.g., carbohydrates, lipids, phosphate groups) that are covalent components of the polypeptide or nucleic acid (e.g., attached to the polypeptide or nucleic acid backbone).

[0286] I. Malaria Malaria is a mosquito-borne infectious disease caused by the single-celled eukaryotic parasite Plasmodium, transmitted through the bite of mosquitoes of the genus Anopheles (Phillips, M., et al. Malaria. Nat Rev Dis Primers 3, 17050, 2017, the entire text of which is incorporated herein by reference). Mosquitoes that transmit malaria must have been infected by having previously ingested blood from an infected subject (e.g., a human). When a mosquito bites an infected subject, it ingests a small amount of blood containing the malaria parasite. The infected mosquito can then bite an uninfected subject and infect them.

[0287] Malaria remains one of the most serious infectious diseases, causing approximately 200 million clinical cases and 500,000 to 600,000 deaths annually. While considerable effort has been made to develop treatments for malaria, many malaria parasites have developed resistance to available therapies. According to the Malaria Eradication Research Agenda Initiative, malaria eradication is only achievable through effective vaccination.

[0288] In 2015, the European Medicines Agency conducted a positive review of a malaria vaccine candidate known as "RTS,S," a landmark in malaria vaccine development. In 2019, the World Health Organization launched a pilot program in parts of three sub-Saharan African countries to provide RTS,S to children at least 5 months old. RTS,S / AS01 is an adjuvant-added protein subunit vaccine consisting of a portion of the main repeat region and the C-terminus of Plasmodium falciparum-derived CSP fused to hepatitis B surface antigen (HBsAg). The vaccine is a mixture of this PfCSP-HBsAg compound and HBsAg that forms virus-like particles (RTS,S / AS01; Mosquirix®). RTS,S is administered according to a regimen requiring four doses: the first three doses given at least one-month intervals, and a fourth dose 15–18 months after dose 3 (see, e.g., 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 have shown that RTS,S protects approximately 30–50% 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, e.g., Moris et al. Hum Vaccin Immunother 14:17, 2018, which is incorporated herein by reference in its entirety). A Phase III trial of RTS,S, delivered as a three-dose series with a booster after one year, demonstrated moderate vaccine efficacy in children aged 5 to 17 months, preventing 36% of clinical malaria cases throughout the study period, with a median follow-up period of 4 years, ranging from 20% in the high-transmission setting to 66% in the low-transmission setting. Furthermore, published literature suggests that protection weakens over time, including reports of potential negative efficacy at 5 years 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).Therefore, an effective malaria vaccine is a critically important and unmet medical need for global health.

[0289] A. Lifecycle During blood-feeding, infected mosquitoes inject sporozoites, known as the hepatic stage of Plasmodium species, along with anticoagulant saliva. The sporozoites pass through the skin and, if successful, enter the hepatocytes of the liver. This process is extremely rapid; it can be completed in just a few minutes (Sinnis et al., Parasitol Int. 2007 Sep;56(3):171-8, the whole is incorporated herein by reference). This is known to be a stage where the host immune system has little opportunity to initiate an effective response against the parasites, as only a small number (perhaps up to several hundred) of sporozoites are injected by the mosquito, and only a small fraction of these parasites establish infection in the liver (Flores-Garcia et al., mBio. 2018 Nov 20;9(6):e02194-18, the whole is incorporated herein by reference).

[0290] Once sporozoites migrate from the skin inoculation site to the liver, they traverse host cells (Mota et al., Science 2001 Jan 5;291(5501):141-4). To gain access to hepatocytes, sporozoites traverse various types of host cells in the dermis, including fibroblasts and phagocytes (Amino et al., Cell Host Microbe. 2008 Feb 14;3(2):88-96, the whole of which is incorporated herein by reference), as well as the hepatic sinusoidal barrier, which contains hepatic endothelial cells and Kupffer cells (Frevert et al., PLoS Biol 3(6):e192. 2005, the whole of which is incorporated herein by reference) and sinusoidal endothelial cells (Tavares et al., J Exp Med 2013 May 6;210(5):905-15, the whole of which is incorporated herein by reference). Sporozoites preferentially cross cells containing low-sulfated heparin sulfate proteoglycans (HSPGs), but preferentially invade cells containing high-sulfated HSPGs (Coppi et al., Cell Host & Microbe 2, 316-327, November 2007, the entire paper is incorporated herein by reference).

[0291] Cellular transposition was initially observed as non-phagocytic invasion of mouse malaria parasite (P. berghei) sporozoites into macrophages, followed by “escape” from these cells (Vanderberg et al., J.Euk.Microbiol.37:528-536, 1990, the whole is incorporated herein by reference). Biochemical, biophysical, and stepwise transposition processes are still being investigated. However, electron microscopy has suggested that host cell rupture occurs during host cell entry and exit (Mota et al., 2001; Tavares et al., 2013, each of the whole is incorporated herein by reference). Furthermore, it has been shown that *P. yoelii* sporozoites can enter hepatocytes via transient vacuoles, and that host membrane rupture occurs during cell exit rather than cell entry (Risco-Castillo et al., Cell Host Microbe 2015 Nov 11;18(5):593-603, the entire paper is incorporated herein by reference).

[0292] Sporozoites also traverse hepatocytes before establishing proliferative hepatocyte infection (Mota et al., 2001, the whole is incorporated herein by reference). Several possibilities have emerged regarding why this occurs. The first hypothesis suggested that migration through hepatocytes evokes the parasite for invasion by activating apical exocytosis (Mota et al., Nat Med 2002 Nov;8(11):1318-22, the whole is incorporated herein by reference). The second theory suggested that traversal releases hepatocyte growth factor (HGF), making adjacent hepatocytes more susceptible to infection (Carrolo et al., Nat Med. 2003 Nov;9(11):1363-9, the whole is incorporated herein by reference). Finally, other studies suggest that sporozoites take some time to switch off the transverse mechanism and activate the entry mechanism (Amino et al., 2008, Coppi et al., 2007, each of which is incorporated herein by reference in whole), and that transverse primarily functions to bypass phagocytosis in the pathway to the liver by penetrating the cell barrier (Amino et al., 2008, Coppi et al., 2007, Tavares et al., 2013, each of which is incorporated herein by reference in whole).

[0293] Sporozoites have been shown to cross 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, each of these is incorporated herein by reference in whole), but the molecular basis of the cross-sectional process has been largely unstudied. Antibodies against circum sporozoite protein (CSP) impair cell crossing (Dumoulin et al., 2015, the whole article is incorporated herein by reference), but this is likely due to inhibition of motility rather than a direct effect (Cha et al., J Exp Med 2016 Sep 19;213(10):2099-112, the whole article is incorporated herein by reference). Furthermore, antibodies induced by CPS immunization (chloroquine prophylaxis with sporozoites) can target CSP and interfere with cell crossing (Behet et al., 2014, the whole article is incorporated herein by reference). Recently, it has been shown that glyceraldehyde 3-phosphate dehydrogenase (GAPDH) on the surface of parasites interacts with CD68 on Kupffer cells during crossing (Cha et al., 2015, Cha et al., 2016, each of these is incorporated herein by reference in its entirety).

[0294] In rodent malaria parasites such as the mouse malaria parasite, two sporozoite microname proteins have been identified that appear to be essential for cell translocation (Sporozoite microname protein essential for cell translocation [Spectrum 1; Ishino et al., PLoS Biol., 2 (2004), pp. 77-84, the whole is incorporated herein by reference] and SPECT2 [Ishino et al., Cell. Microbiol., 7 (2005), pp. 199-208, the whole is incorporated herein by reference], also known as perforin-like protein 1 [PLP1] [Kaiser et al., Mol. Biochem. Parasitol., 133 (2004), pp. 15-26, the whole is incorporated herein by reference]. Even when gene disruption of SPECT1 or SPECT2 prevented sporozoites from translocating mouse cells, they still entered hepatocytes in vitro (Ishino et al. (Ishino et al., 2004, Ishino et al., 2005; each of these is incorporated herein by reference in its entirety). When injected into rodents, sporozoites lacking SPECT1 or SPECT2 were impaired for liver infection, although a small number of sporozoites were still able to establish liver infection, leading to subsequent manifestation. However, due to Kupffer cell depletion, mutants were able to establish liver infection at a level comparable to that of wild-type parasites (Ishino et al., 2004, Ishino et al., 2005; each of these is incorporated herein by reference in its entirety). This data suggests that crossing by rodent-infectious sporozoites is important for navigation through the sinusoidal layer, but not for hepatocyte invasion, development of malarial exteroerythrocyte morphology, or growth within erythrocytes (Ishino et al., 2004, Ishino et al., 2005; each of these is incorporated herein by reference in its entirety).

[0295] PLP1, an ortholog of SPECT2 in *Parasomalis yoweri*, has been shown to play a role in cell transposition. This protein is not required for hepatocyte entry but plays a role in exiting transient vacuoles during transposition (Risco-Castillo et al., 2015, each of which is incorporated herein by reference in whole). Therefore, sporozoites infecting rodents may transposition host cells by generating vacuoles in the entry step and use perforin-like proteins (e.g., SPECT2 / PLP1) to escape this compartment and / or from the host cell during cell exit.

[0296] When sporozoites invade liver cells, they differentiate into merozoites, the replicated form of the parasite. Within days, a single sporozoite can give rise to 5,000 to 10,000 merozoites. The merozoites sprout from host liver cells in structures called merosomes, which contain up to 1,000 merozoites and are hidden from the host immune response due to their host membrane composition. These merosomes eventually release merozoites that freely mix with the host cytoplasm and appear in the bloodstream, where they invade red blood cells and initiate the blood phase of infection, characterized by rapid asexual replication and clinical symptoms. Within days, hundreds of thousands of parasites can be present in the bloodstream.

[0297] Plasmodium parasites invade red blood cells via specific ligand-receptor interactions mediated by proteins on the surface of the parasite that interact with receptors on host red blood cells (mature red blood cells) or reticulocytes (immature red blood cells). Plasmodium falciparum can invade and replicate in both red blood cells and reticulocytes, while Plasmodium vivax and other species primarily invade reticulocytes, which are fewer in number than red blood cells. The majority of red blood cell-binding proteins or reticulocyte-binding proteins involved in invasion are either duplicated or expressed as families of variant forms; however, for Plasmodium falciparum, two essential red blood cell receptors (basidine and complement-disintegrating factor (also known as CD55)) have been identified.

[0298] Plasmodium vivax and Plasmodium ovale can enter a dormant state in the liver, which is a hypnozoite.

[0299] Merozoidal invasion of red blood cells involves interactions between multiple parasite-derived proteins (e.g., ligands) and red blood cell (RBC) proteins (e.g., receptors) (Weiss et al., Plos Path., 2015 Feb 27;10.1371, the whole is incorporated herein by reference). The invasion process begins with the initial attachment of merozoites to RBCs. Merozoidal-RBC interactions are further enhanced through merozoite surface protein-1 (MSP1) and an unknown RBC protein, causing some deformation of the RBC surface. Following deformation, interactions between merozoite EBA and the Rh protein family (excluding Rh5) and host receptors such as CR1 result in actin-dependent deformation of the RBC membrane and reorientation of the merozoite tip onto the RBC surface (Geoghehan et al., Nat Commun., 2021 June 15;10.1038, the whole is incorporated herein by reference). This deformation event increases the surface area of ​​the interaction between the merozoite and the RBC membrane, and this interaction can be stabilized by an Rh5 entry complex containing plasmodium thrombospondin-associated apical merozoite protein (PTRAMP), cysteine-rich microsecretion (CSS), cysteine-rich protective antigen (CyRPA), Rh5 interacting protein (Ripr), and Rh5. This Rh5 entry complex, containing five proteins, is called the PCRCR complex.

[0300] The formation of the PCRCR complex begins in the endoplasmic reticulum, where its components, the cysteine-rich small secretory (CSS) protein and the plasmodium thrombospondin-associated apical merozoite protein (PTRAMP), interact to form a heterodimer called PTRAMP-CSS. PTRAMP-CSS is transported to secretory organelles called micronems, where CSS interacts with the Rh5 interacting protein (Ripr) and the cysteine-rich protective antigen (CyRPA) to form a tetrameric complex called PCRC. At some point after deformation occurs, the merozoite initiates a polar secretory process, which allows the micronemal PCRC complex to interact with the rh5 protein, forming the pentameric complex PCRCR. Subsequently, through Rh5, the PCRCR complex can bind to a host cell receptor called basidine, broadly speaking, throughout the merozoite-RBC interphase. The binding of PCRCR to basidine creates a stable and irreversible platform between the merozoite tip and the deformed RBC surface, enabling the embedding of Rh5 and Ripr into the RBC membrane (Scally et al., Nat Microb., 2022 May 4, the whole is incorporated herein by reference). Once the merozoite-RBC interaction is established, an open connection is formed between the merozoite tip and the RBC surface, which is Ca 2+ It acts as a conduit for merozoite-derived invasion proteins such as AMA1 and RON2, to facilitate their influx into the RBC and to help establish a migratory junction (Srinivasan et al., Proc Natl Acad Sci., 2011 Jul 25;13275-80, the whole of which is incorporated herein by reference). Through this migratory junction, the parasite uses its actin-myosin-dependent gliding motility to propel itself within the RBC and establish a parasitic vacuole from which the parasite grows and replicates.

[0301] Rh5 has been shown to bind to basidine with higher affinity when combined with CyRPA and Ripr to form an RCR complex, and to have the highest affinity for basidine when it is part of a PCRCR complex (Wong et al., Nature, 2019 and Scally et al., Nat Microb., May 4, 2022, each of which is incorporated herein by reference in whole).

[0302] Merozoids that escape from red blood cells can invade other red blood cells and continue into the asexual hematopoietic phase of the parasite's life cycle. A small percentage of schizonts are already destined for a different fate, and merozoites from these schizonts differentiate into either male or female gamete cells in the parasite's life cycle after invasion of new red blood cells. Gamete cells are taken up by mosquito vectors during blood supply. In the mosquito's intestines, male gamete cells undergo three rapid mitotic divisions to form eight flagellated microgametes. Female gamete cells mature into macrogametes that escape from their red blood cells. Male microgametes are the flagellated, motile form that seeks female gametes. Male and female gamete cells fuse to form a diploid zygote, which matures and differentiates into okinates; this motile form secretes chitinase and other soluble proteins such as CelTOS to disrupt and traverse the peritrophic matrix, cross the midgut epithelium, and reach the basement membrane, where it further differentiates and matures as an oocyst. The oocyst matures and replicates over approximately 10 days (depending on temperature), forming sporozoites that escape from the mature oocyst into the mosquito's blood clot. Thousands of sporozoites are formed from a single oocyst, becoming randomly distributed throughout the hemocall. The sporozoites are thought to passively circulate through the mosquito's hemolymph until they encounter a salivary gland, where they actively invade it. After invading the salivary gland, the sporozoites are reprogrammed via an unknown mechanism to prepare for liver invasion. Evidence for this reprogramming is also demonstrated by the fact that midgut sporozoites (directly from oocysts) cannot invade liver cells, and that sporozoites that successfully invade a salivary gland cannot reinvade another salivary gland when presented. Salivary gland sporozoites alter mosquito behavior and salivary gland function as the amount of saliva produced decreases, resulting in increased mosquito foraging behavior and a higher likelihood of transmission to a human host through mosquito bites, thus continuing the human host stage of this parasite's life cycle.

[0303] Plasmodium vivax and Plasmodium ovale can enter a dormant state in the liver, which is a hypnozoite.

[0304] One specific challenge associated with initiating an immune response that can clear sporozoites before liver infection is established is that circumsporozoite protein (CSP), the most abundant and immunogenic protein on the sporozoite surface, is exposed to the immune system only in small amounts and for short periods due to the varyingly low inoculation amounts from mosquitoes and the dynamics of hepatocyte infection after inoculation. Furthermore, after liver infection is established, the parasites differentiate into stages that no longer express CSPs and instead have a different mosaic of surface antigens, making the parasites invisible to any memory immune response directed, for example, towards the liver phase. The initial stages of parasitic exposure to the human host create a short window for the innate and memory immune systems to generate a response that can clear all parasites before infection is established. The clinical significance of immune evasion by sporozoites is high at this stage because only a single sporozoite is needed to produce as many as 10,000 hematopoietic parasites (merozoites) that can invade hepatocytes, invade red blood cells, replicate within them, and lyse (Shears et al., jproteome. 2019 July 23;3404-3418, the whole is incorporated herein by reference). These early stages of the parasitic life cycle in the human host help to demonstrate the difficulty of targeting hematopoietic parasites for therapeutic intervention and the low efficacy seen with current vaccines (Oluta A. et al., N Engl J Med. 2016 June 30;216:374, the whole is incorporated herein by reference). Therefore, targeting the hematopoietic stage of Plasmodium infection offers a promising approach to reduce mortality caused by malaria.

[0305] Malaria symptoms typically develop 4 to 8 days after the initial red blood cell infestation. The merozoite replication cycle within red blood cells continues for 36 to 72 hours before hemolysis occurs, releasing merozoites for another red blood cell infection. Therefore, in co-infections (infections originating from a single infectious bite), fever occurs every 36 to 72 hours as infected red blood cells lyse in large quantities and release endotoxins.

[0306] Several drugs that prevent the entry or proliferation of Plasmodium species into the liver have prophylactic activity, drugs that block the erythrocyte phase are necessary for treating the symptomatic stage of the disease, and compounds that inhibit the formation or development of gamete cells in mosquitoes (including drugs that kill blood-feeding mosquitoes) are signaling blockers (Phillips, et al. Malaria. Nat Rev Dis Primers 3, 17050 (2017), the whole of which is incorporated herein by reference).

[0307] B genome Since the initial sequencing of the Plasmodium falciparum 3D7 genome was completed in 2002, genomic research on malaria parasites has advanced rapidly. Except for a short diploid stage after fertilization in the midgut of mosquitoes, Plasmodium parasites are haploid throughout their life cycle. Genomes of different species range from 20 to 35 megabases and contain 14 chromosomes, approximately 35 kilobases of circular plastid genome, and multiple copies of 6 kilobases of mitochondrial DNA. Comparisons of genomes from different species have shown that homologous genes are often found in synthetic blocks arranged in different orders between different chromosomes.

[0308] The adenine-thymine (AT) content of Plasmodium species can also vary considerably, for example, around 80% AT in Plasmodium falciparum, Plasmodium reichenowi, and Plasmodium gallinaceum; around 75% AT in rodent malaria parasites; and around 60% AT in Plasmodium vivax, Plasmodium knowlesi, and Plasmodium cynomolgi. AT content is often higher in introns and intergeneric non-coding regions than in protein-coding exons, with an average of 80.6% AT across the entire Plasmodium falciparum genome compared to 86.5% in non-coding sequences. The high AT content of Plasmodium falciparum reflects numerous low-complexity regions, simple sequence repeats and microsatellites, as well as a highly skewed codon use frequency bias. AT-rich repeat polymorphisms provide a wealth of markers for linkage mapping of drug resistance genes and for tracking the evolution and structure of parasite populations.

[0309] The genomes of malaria parasites contain a multi-gene family that plays a crucial role in their interactions with their hosts, including antigenic mutation, signaling, protein transport, and adhesion. Among these gene families, the gene encoding Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) is the most widely studied. Each Plasmodium falciparum parasite possesses a unique set of 50–150 copies of the var gene in its genome, and the switching of gene expression can lead to antigenic mutation. PfEMP1 plays a vital role in the pathogenesis of clinical manifestations such as cerebral malaria and placental malaria, mediating the cell adhesion of infected erythrocytes (iRBCs) in deep tissues. Different PfEMP1 molecules bind to a variety of host molecules, including α2-macroglobulin, CD36, chondroitin sulfate A (CSA), complement 1q, CR1, E-selectin and P-selectin, endothelial protein C receptor (EPCR), heparan sulfate, ICAM1, IgM, IgG, PECAM1, thrombospondin (TSP), and VCAM1. Such binding leads to the activation of various host inflammatory responses. Abnormal hemoglobinopathy, including hemoglobin C and hemoglobin S phenotypes, interferes with PfEMP1 display in the knob structure of iRBCs. This inadequate display of PfEMP1 on the host cell surface provides protection against malaria by reducing cell adhesion and activation of inflammatory processes that promote the development of severe disease.

[0310] Members of the large plasmodium sporadic repeat (pir) multiple gene family are named differently depending on the parasite species; for example, yir in Plasmodium erii, bir in Plasmodium biennis, and vir in Plasmodium vivax. Several Plasmodium falciparum gene families (Stevar, rif, and PfMC-2™) are classified as pir by their similar gene structure, which characteristically contains a short first exon, a long second exon, and a third exon encoding a transmembrane domain. Recent studies have shown that the pir gene (cir) from Plasmodium chabaudii is expressed inside and on the surface of iRBCs, as well as at different cellular locations within merozoites. Malaria parasites allocate a large portion of their genome to gene families that ensure evasion of host immune defenses and protection of molecular processes essential for infection. These families highlight the importance of studying their role in parasite-host interactions and pathogenicity, despite the inherent difficulties in investigating them.

[0311] Further exemplary polymorphic gene families include a group of 14 genes encoding proteins containing six cysteine ​​(6-Cys). These proteins often localize to the parasite surface, interacting with host proteins, and are expressed at different stages of parasite development. 6-Cys proteins have also demonstrated diverse functions, and have been shown to play roles, for example, in parasite fertilization, conjugation interactions, evasion of immune responses, and hepatocyte invasion. Proteins expressed during the asexual phase are generally polymorphic and / or selective, suggesting they may be targets of host immune responses; however, their function in parasite development remains largely unknown.

[0312] Plasmodium genomes can be highly polymorphic. Early studies have demonstrated polymorphisms involving tens to hundreds of kilobases, showing that while the chromosomal structure of Plasmodium falciparum is largely conserved in the central region, both telomere length and sequence are extensively polymorphic. Many subtelomere variations have been explained by blocks of repetitive sequences and recombination within gene families.

[0313] The frequency of simple sequence repeats (microsatellites) in Plasmodium falciparum is estimated to be approximately one polymorphic microsatellite per kbDNA. While we do not wish to be bound by any one theory, this high proportion may reflect the AT-rich nature of the genome. Microsatellites appear to be less frequent in other Plasmodium species with genomes having lower AT content. In addition to the highly polymorphic and repetitive structures of the Plasmodium genome, numerous single nucleotide polymorphisms (SNPs) and copy number variations (CNVs) are also present (Su et al., Plasmodium Genomics and Genetics: New Insights into Malaria Pathogenesis, Drug Resistance, Epidemiology, and Evolution. Clin Microbiol Rev. 2019 Jul 31;32(4), the entire text is incorporated herein by reference).

[0314] C. plasmodium protein 1. Rh5 Invasion Complex Polypeptide Rh5 Rh5 is found in Plasmodium falciparum, but not in all other species of Plasmodium that infect humans that contain proteins with similar functions to Rh5. Rh5 orthologs are also found in other species belonging to the subgenus Lavalenia, including parasites that infect chimpanzees and gorillas, and they play a unique role in Plasmodium falciparum invasion of human red blood cells. See, for example, Ragotte, et al. Trends Parasitol. 36(6) 2020, the entire work of which is incorporated herein by reference. Rh5 is expressed during the mature schizont stage and is localized to loptri, the polar secretory organelle of Plasmodium falciparum. Rh5 is a protein secreted during the invasion process and is thought to be essential for the completion of parasitic invasion of red blood cells. Rh5 functions downstream of the initial attachment of the parasite to the red blood cell surface and deformation of the red blood cell membrane. Upon secretion, Rh5 binds to the erythrocyte surface protein basidine, creating an adhesion and stable platform for downstream receptor-ligand interactions and associated entry events. Rh5 binding to basidine is also necessary for inducing calcium spikes within the erythrocyte, which are inhibited if merozoites attempt to enter in the presence of anti-Rh5, anti-Ripr, or anti-basidine antibodies or soluble basidine (see, for example, Ragotte (2020), whose entire work is incorporated herein by reference).

[0315] Rh5 can interact with multiple proteins to form complexes. The affinity of Rh5 for basidine varies depending on which Rh5 complexes are present. For example, Rh5 can form an elongated protein trimer called RCR on the merozoite surface, which binds to the erythrocyte surface protein basidine with higher affinity than Rh5 alone. (See, for example, Ragotte (2020) and Wong (2019), each of which is incorporated herein by reference in whole.) Rh5 has the highest affinity for basidine when it interacts with PTRAMP, CSS, Ripr, and CyRPA in a pentameric complex known as PCRCR. See Scally (2022).

[0316] Rh5 is a 63 kDa protein expressed during the mature schizont stage. It is processed, cleaved into a 45 kDa form, and released by the parasite. The structure of Rh5 reveals a kite-like structure formed by the joining of two triplicate bundles. See, for example, Ragotte (2020), the whole of which is incorporated herein by reference.

[0317] The Rh5 sequences are publicly known (e.g., UniProt accession numbers A0A159SK44, A0A159SK99, A0A159SKS8, A0A159SKW8, A0A159SL23, A0A159SL78, A0A159SL96, A0A159SLM7, A0A159SMC8, A0A159SMR9, A0A161FQT0, A0A1B1UZE2, A0A1B1UZE4, A0A See 1B1UZE5, A0A346RCI1, A0A346RCJ0, A0A346RCJ2, A0A346RCJ3, A0A346RCJ4, A0A346RCK4, A0A346RCK5, A0A346RCK6, A0A346RCK9, B2L3N7, Q8IFM5 (each of which is incorporated herein by reference in whole), exemplary Rh5 amino acid sequences are provided in Table 1.

[0318] Rh5-interacting protein (Ripr) The Rh5-interacting protein (Ripr) is a protein of approximately 120 kDa that localizes to the micronome during the schizont phase of the plasmodium life cycle. The full-length 120 kDa protein is processed into two fragments of similar size: an N-terminal fragment (containing EGF domains 1 and 2) and a C-terminal fragment (containing EGF domains 3–10). Ripr co-localizes with Rh5 and CyRPA during parasitic invasion at the junction between merozoites and erythrocytes. Parasites with conditional knockout of PfRipr induce membrane deformation but are unable to achieve complete invasion (see, e.g., Ragotte (2020), whose entire work is incorporated herein by reference).

[0319] Ripr sequences are publicly known (see, for example, UniProt accession numbers A0A193PDI9, A0A193PDK3, A0A193PDK8, A0A193PDL3, A0A193PDL9, A0A193PDP4, A0A193PDQ8, A0A193PE01, A0A193PE05, A0A193PE07, O97302, and A0A193PE17). Exemplary Ripr amino acid sequences are provided in Table 1.

[0320] Cysteine-rich protective antigen (CyRPA) Cysteine-rich protective antigen (CyRPA) is a 43 kDa protein with a predicted N-terminal secretory signal. CyRPA is a key mediator in the invasive multiprotein complex known as RCR, which consists of Rh5 and Ripr, and improves the Rh5 affinity to the basidine receptor on erythrocytes. The ability of CyRPA to make limited contact with Ripr and Rh5 is thought to help further stabilize the contact between parasites and erythrocytes by enabling the dissociation of Rh5 and Ripr from this complex, allowing them to be inserted into the erythrocyte membrane. CyRPA also plays a role in the PCRCR complex, which further increases the Rh5 affinity to basidine. CyRPA-mediated Rh5 binding to basidine is linked to Ca2 in the erythrocyte cytosol. +It is important for stabilizing parasite-erythrocyte membrane interactions, which enable downstream invasion events such as increased activity and establishment of tight / mobile junctions. CyRPA is highly conserved, with only a single SNP present in over 5% of the general population. CyRPA is important for invasion (conditional knockdown causes loss of invasion activity), and CyRPA has low serological reactivity from natural exposure (see, e.g., Ragotte (2020) and Cowman (2018), each of which is incorporated herein by reference in whole).

[0321] CyRPA sequences are publicly known (see, for example, Uniprot accession numbers A0A2S1Q7P0, A0A2S1Q7P5, A0A2S1Q7Q4, Q8IFM8, each of which is incorporated herein by reference in its entirety). Exemplary CyRPA amino acid sequences are provided in Table 1.

[0322] Plasmodium thrombospondin-associated apical merozoite protein (PTRAMP) Plasmodium thrombospondin-associated apical merozoite protein (PTRAMP) is a 352-amino acid protein that localizes to developing micronomes and relocalizes to the merozoite surface upon invasion, and is understood to be crucial for Plasmodium falciparum's invasion of host cells. Orthologues of the gene encoding PTRAMP exist in all malaria parasite species investigated, demonstrating a conserved role in host invasion and making it a convenient target for therapeutic intervention. Important for a vaccine approach, PTRAMP appears to exhibit both linear and conformational epitopes, which gives the immune system the opportunity to utilize both B-cell and T-cell receptors for recognition and activation upon exposure to PTRAMP (Alvaro et al., Malaria Jou., 2010 Oct. 13; 10.1186, the entire text is incorporated herein by reference). PTRAMP can interact with cysteine-rich microsecretory (CSS) proteins to form a heterodimer known as PTRAMP-CSS, which, through CSS, can interact with Ripr and CyRPA to form a tetrameric complex called PCRC, and finally interact with Rh5 to form a pentameric complex PCRCR, which enhances Rh5 binding to host basidine, crucial for Plasmodium falciparum entry into RBCs. Nanobodies of PTRAMP inhibit merozoites, significantly reducing merozoite entry and providing evidence of the potential efficacy that therapeutically targeting PTRAMP may offer (Scally et al., Nat Microb., 2022 May 4, the whole is incorporated herein by reference).

[0323] The PTRAMP sequence is publicly known (see, for example, Uniprot accession number [Q8I5M8]). An exemplary PTRAMP amino acid sequence is provided in Table 1.

[0324] Cysteine-rich microsecretory protein (CSS) Cysteine-rich microsecretory proteins (CSS) are 290-amino acid latent 6-Cys proteins composed of two degenerate 6-Cys domains. Proteins belonging to this family generally mediate extracellular protein-protein interactions, a role that has been confirmed to occur in Plasmodium falciparum. CSS can bind to both PTRAMP and Ripr. CSS interactions with PTRAMP occur in the endoplasmic reticulum, while CSS interactions with Ripr occur after transport to the micronome. CSS interactions with PTRAMP and Ripr are involved in entry and the formation of the PCRCR complex, which can complex with Rh5 and enhance the interaction of Rh5 with basidine. Nanobodies binding to CSS, similar to PTRAMP, have been shown to inhibit entry, suggesting that its role in the entry process is likely its involvement in the PCRCR complex. Nanobodies inhibiting CSS binding to PTRAMP or Ripr do not inhibit entry, confirming the interaction of these proteins before exposure to the merozoite surface. The inhibitory activity of nanobodies against CSS is thought to arise by potentially blocking the insertion of Ripr and / or Rh5 into the RBC membrane and / or disrupting the conformational changes of the PCRCR complex necessary for entry, although the exact mechanism of inhibition is not fully understood. In contrast to this observation and its immunological significance, polyclonal antibodies against CSS and PTRAMP exhibit no neutralizing activity, indicating immunodominance of non-neutralizing epitopes of CSS and PTRAMP, which biases antibody selection from neutralizing epitopes (in the natural environment). This observation provides a molecular basis for identifying entry-inhibiting epitopes on CSS and PTRAMP that are highly conserved in Plasmodium falciparum for use in vaccine platforms. See Cowman (2022).

[0325] The CSS sequence is publicly known (see, for example, Uniprot accession number [Q8IM47]). An exemplary CSS amino acid sequence is provided in Table 1.

[0326] In some embodiments, the plasmodium CSS polypeptide or its antigenic moiety contains serine at position 30, numbered according to SEQ ID NO: 5. Exemplary amino acid sequences of CSS including the C30S mutation are provided in Table 1.

[0327] P113 P113 is a glycosylphosphatidylinositol (GPI)-binding protein that directly interacts with the N-terminus of raw Rh5, and it has been thought that this provides a mechanism for the Rh5 entry complex to tether to the merozoite surface. However, recent findings suggest that P113 may not be essential for Plasmodium falciparum growth, as other proteins and complexes exist that can perform P113's Rh5 membrane tethering function. See, for example, Ragotte (2020) and Scally (2022). In addition to its canonical role, P113 has been shown to be present in the paristopherous vacuole (PV) and interact with parasitic secretory complexes such as PTEX and EPIC, which are important for the secretion of parasitic effector proteins across the paristopherous vacuole membrane (PVM). P113 has also been shown to play a role in the PVM architecture, as deletion of this gene results in disruption of PVM morphology. P113 orthologues have been found in all Plasmodium species sequenced to date, suggesting a common conserved function (Bullen et al. (2022) Molecular Microbiology 117:1245-1262, the whole is incorporated herein by reference). Despite this fact, in mouse malaria parasites, P113 knockout parasites were viable, demonstrating that this protein is not essential for growth and entry during the asexual reproductive hematopoietic phase. However, the knockout parasites exhibit defects in natural sporozoite transmission, resulting in delayed manifestation in infected mice (Offeddu et al. (2014) Mol. Biochem. Parasitology 193:101-109, the whole is incorporated herein by reference).

[0328] The plasmodium P113 sequence is publicly known (see, for example, Uniprot accession number Q8ILP3). An exemplary P113 amino acid sequence is provided in Table 1.

[0329] [Table 1]

[0330] [Table 2]

[0331] D. Further plasmodium proteins Plasmodium parasites are known to express various proteins at different stages of their life cycle. Exemplary malaria proteins are described below, and their exemplary amino acid sequences are provided in Table 2.

[0332] Circum sporozoite protein (CSP) Circum sporozoite proteins (CSPs) are multifunctional proteins involved in the life cycle of Plasmodium, as they are required for sporozoite formation in the midgut of mosquitoes, sporozoite release from oocysts, entry into salivary glands, attachment of sporozoites to hepatocytes in the liver, and sporozoite entry into hepatocytes (see, for example, Zhao et al. (2016) PLoS ONE11(8):e0161607, which is incorporated herein by reference in its entirety). CSPs are present in all Plasmodium species, and while there are amino acid sequence variations between species, the overall domain structure of the central repeat region and non-repeat facies is well conserved (see, for example, Zhao et al. (2016) PLoS ONE11(8):e0161607; Wahl et al. (2022) J.Exp.Med.219:e20201313, each of which is incorporated herein by reference in its entirety). CSP sequences are publicly known (see, for example, UniProt accession numbers A0A2L1CF52, A0A2L, 1CF88, C6FGZ3, C6FH2,7 C6FHG7, M1V060, M1V0A3, M1V0B0, M1V0C4, M1V0E0, M1V9I4, M1VFN9, M1VKZ2, P02893, Q5EIJ9, Q5EIK2, Q5EIK8, Q5EIL3, Q5EIL5, Q5EIL8, Q5R2L2, Q7K740, Q8I9G5, Q8I9J3, Q8I9J4). Exemplary CSP amino acid sequences are provided in Table 2.

[0333] E140 E140 is found in all Plasmodium species whose genome sequences are available, are well-conserved, and exhibit interspecies amino acid identity ranging from 34% to 92%. See, for example, Smith, et al. PLoS one 15.5(2020):e0232234;http: / / doi:10.1371 / journal.pone.023223; and U.S. Patent Application Publication 2019 / 0117752, each of which is incorporated herein by reference in whole. E140 is also highly conserved (95–99%) in Plasmodium falciparum strains isolated from different locations around the world and exhibits a low mutation frequency. E140 is expressed at different life stages of malaria parasites (specifically, E140 has been detected in sporozoite, hepatic, and hematopoietic stage parasites).

[0334] Protein structure algorithms predict that the E140 protein has five transmembrane domains that are presumed to extend to parasite or host-derived membranes. E140 exhibits different patterns of protein expression in mature sporozoites, late hepatic, and late schizontic stages. It migrates around the anterior and posterior ends of sporozoites, the parasitic vesicle space in the late hepatic stage, and the developing merozoites in the late schizontic stage. It is also known to be expressed in mature salivary gland sporozoites and oocyst-derived sporozoites and oocysts.

[0335] The E140 sequence is publicly known (see, for example, UniProt accession numbers A0A650D649, A0A650D653, A0A650D672, A0A650D687, A0A650D690, A0A650D694, A0A650D6A3, A0A650D6B8, A0A650D6L3, A0A650D6L7, Q8I299, each of which is incorporated herein by reference in its entirety), and exemplary E140 amino acid sequences are provided in Table 2.

[0336] CelTOS CelTOS is required for sporozoite crossing through Kupffer cells during the liver invasion process. CelTOS forms pores from within the cell, enabling sporozoite escape into the liver. Antibody epitopes have been characterized from immunized mice and infected human populations (Pf and Pv). Mouse studies have shown that immunization with CelTOS provides protection and defense against antigenic stimulation. Vaccination with CelTOS may generate antibodies that can bind to the extracellular domain of the pore-forming complex, blocking complete pore formation and preventing sporozoite crossing into the liver. See, for example, Jimah et al., Elife 2016 Dec 1;5:e20621.doi:10.7554 / eLife.20621, which is incorporated herein by reference in its entirety.

[0337] The CelTOS sequence is publicly known (see, for example, Uniprot accession numbers M1ETJ8, Q53UB7, A0A2R4QLA5, A0A2R4QLI0, A0A2R4QLI5, A0A2R4QLJ1, A0A2R4QLJ4, M1ETJ8, Q53UB8, Q8I5P1, each of which is incorporated herein by reference in its entirety). Exemplary CelTOS amino acid sequences are provided in Table 2.

[0338] SPECT1 and SPECT2 SPECT1 and SPECT2 (the latter sometimes referred to as perforin-like protein 1 (PLP1)) are important plasmozium proteins that may play a role in cell translocation. See Yang et al., Cell Rep. 2017 Mar 28;18(13):3105-3116.doi:10.1016 / j.celrep.2017.03.017, which is incorporated herein by reference in its entirety. Targeted disruption of Plasmodium falciparum SPECT1 or SPECT2 has been shown to reduce sporozoite infectivity during hepatic development in humanized mice. However, the cell translocation mechanisms of these two proteins have not yet been defined in Plasmodium falciparum. See Yang et al.

[0339] SPECT1 and SPECT2 are considered attractive pre-erythrocyte immune targets due to their important roles thought to play in the traverse of malaria parasites across the dermis and hepatic sinusoidal walls before hepatocyte invasion. Recombinant Plasmodium falciparum SPECT2, like the MACPF / CDC domain of PfSPECT2, has Ca 2+ It has been shown to induce erythrocyte lysis in a dependent manner. PfSPECT2 is also involved in the Ca2+-dependent extrusion of Plasmodium falciparum merozoites from erythrocytes.

[0340] The SPECT1 and SPECT2 sequences are publicly known (see, for example, UniProt accession numbers Q8IDR4 and Q9U0J9, each of which is incorporated herein by reference in its entirety), and exemplary amino acid sequences of SPECT1 and SPECT2 are provided in Table 2.

[0341] Secretory protein 1 (EXP1) Secretory protein 1 (EXP1) is a single-pass transmembrane protein with an N-terminal signal peptide that is expressed during the intraerythrocyte and hepatic phases (see, for example, Spielmann et al., Int J Med Microbiol. 2012 Oct;302(4-5):179-86, the entire report of which is incorporated herein by reference). EXP1 has been shown to initially localize to high-density granules in merozoites and then be secreted to the parasitic spore membrane (PVM) after entry (see, for example, Iriko et al., Parasitol Int. 2018 Oct;67(5):637-639, the entire report of which is incorporated herein by reference). Upon localization to the PVM, EXP1 forms a homooligomer having an N-terminus exposed to the parasitic sac lumen and a C-terminus exposed to the erythrocyte cytoplasm (see, for example, Mesen-Ramirez et al., PLoS Biol. 2019 Sep 30;17(9):e3000473, the whole of which is incorporated herein by reference).

[0342] EXP1 has been demonstrated to possess glutathione S-transferase (GST) activity that can protect plasmodium from oxidative damage (see, e.g., Mesen-Ramirez et al., PLoS Biol 17(9) 2019 Sep 30;17(9):e3000473, the entire work of which is incorporated herein by reference). More recently, EXP1 has been demonstrated to be important for plasmodium survival by maintaining the precise localization of EXP2, a nutrient permeability channel in PVMs (see, e.g., Mesen-Ramirez et al., PLoS Biol. 2019 Sep 30;17(9):e3000473, the entire work of which is incorporated herein by reference).

[0343] The EXP1 polypeptide sequence of Plasmodium falciparum is publicly known (see, for example, UniProt accession numbers Q8IIF0, W7JTD3, Q25840, Q548U2, Q5VKK2, Q5VKK5, Q5WRH8, Q6V9G4, Q6V9G6, Q6V9G9, Q6V9H1, Q6V9H2, Q9U590, P04923, P04926, each of which is incorporated herein by reference in whole). Exemplary EXP1 amino acid sequences are provided in Table 2.

[0344] Gene 3 (UIS3) is upregulated in infectious sporozoites. Gene 3 (UIS3), which is upregulated in infectious sporozoites, is a membrane-bound protein localized to the sporozoite parasitic spore membrane (PVM) of infected hepatocytes. UIS3 has been shown to interact with hepatic fatty acid-binding protein (L-FABP) and be involved in fatty acid and / or lipid uptake during the plasmodium growth phase (see, for example, Sharma et al., J Biol Chem. 2008 Aug 29;283(35):24077-24088; Mikolajczak et al., Int J Parasitol. 2007 Apr;37(5):483-9, each of which is incorporated herein by reference in whole).

[0345] Following sporozoite infiltration of host hepatocytes, important plasmodium structural features (e.g., parasitic spore membrane) are synthesized. During the hepatocyte phase, plasmodium depends on host fatty acids for the rapid synthesis of its membrane (see, e.g., Sharma et al., J Biol Chem. 2008 Aug 29;283(35):24077-24088, which is incorporated herein by reference in its entirety). UIS3 insertion into the PVM provides a method for introducing essential fatty acids and / or lipids to plasmodium during the rapid sporozoite growth phase (see, e.g., Sharma et al., J Biol Chem. 2008 Aug 29;283(35):24077-24088, which is incorporated herein by reference in its entirety).

[0346] In rodent malaria models, immunization with malaria-protected UIS3-deficient mouse malaria parasite sporozoites is achieved (see, e.g., Mueller et al., Nature. 2005 Jan 13;433(7022):164-7, the entire article of which is incorporated herein by reference). UIS3-deficient mouse malaria parasites can initiate the transformation process in the liver; however, they exhibit severe defects during transformation into trophozoites (see, e.g., Mueller et al., Nature. 2005 Jan 13;433(7022):164-7, the entire article of which is incorporated herein by reference). UIS3-deficient mouse malaria parasites also fail to develop into mature hepatic schizonts and thus prevent malaria infection within the liver itself (see, e.g., Mueller et al., Nature. 2005 Jan 13;433(7022):164-7, the entire article of which is incorporated herein by reference). Furthermore, it has been previously demonstrated that UIS3 derived from *Platypleura malariae* and UIS3 derived from *Platypleura falciparum* exhibit low (i.e., 34%) amino acid sequence identity (see, for example, Mueller et al., Nature. 2005 Jan 13;433(7022):164-7, the entire report of which is incorporated herein by reference).

[0347] Plasmodium UIS3 sequences are publicly known (see, for example, UniProt accession numbers A0A509ARS3, A0A1C6YLP3, Q8IEU1, A0A384KLI1, A0A1G4H423, A0A077YB01, Q9NFU4, each of which is incorporated herein by reference in its entirety). Exemplary UIS3 amino acid sequences are provided in Table 2.

[0348] The early transcription membrane protein 10.3 (ETRAMP10.3) and gene 4 (UIS4) which is upregulated in infectious sporozoites. Gene 4 (UIS4), which is upregulated in infectious sporozoites, contains a single transmembrane domain and is localized to the secretory organelles and hepatic sporozoite membranes (PVMs) of sporozoites. UIS4 is not expressed in hematopoietic or early sporozoites produced in oocysts (see, for example, Mackellar et al., Eukaryot Cell. 2010 May;9(5):784-794, which is incorporated herein by reference in its entirety).

[0349] Deletion of the UIS4 gene is associated with the arrest of early liver development (see, e.g., Vaughan and Kappe, Cold Spring Harb Perspect Med. 2017 Jun 1;7(6):a025486, the entire article of which is incorporated herein by reference). Recently, UIS4 has been demonstrated to be involved in the survival of mouse malaria parasites by evading host actin structures deployed as part of host cytosolic defense (see, e.g., Bana et al., iScience. 2022 Apr 22;25(5):104281.doi:10.1016 / j.isci.2022.104281.eCollection 2022 May 20, the entire article of which is incorporated herein by reference). Plasmodium falciparum has an ortholog for UIS4 named ETRAMP10.3, which is unable to function as a functional complement to Plasmodium yoreris UIS4 and is likely to play a different role in the life cycle of Plasmodium falciparum (see Mackellar et al., Eukaryot. Cell 9:784-94 (2010), which is incorporated herein by reference in its entirety).

[0350] The Plasmodium falciparum early transcription membrane protein 10.3 (ETRAMP10.3) is a protein of approximately 10 kDa and is a member of the Plasmodium falciparum multigene family (a family conserved across Plasmodium species and including proteins located in the parasitic vesicle). Some ETRAMP proteins are specific to Plasmodium falciparum and are not found in Plasmodium species that infect other organisms. ETRAMP10.3 is one such example, expressed in Plasmodium falciparum parasites in both the hepatic and hematopoietic phases. ETRAMP10.3 transcription is known to peak during the transition from the cyclic to the trophozoite phase of Plasmodium falciparum hematopoietic infection in human hosts. ETRAMP10.3 is localized in the parasitic vesicle and secreted into host erythrocytes during hematopoietic infection. ETRAMP10.3 is sometimes described as being upregulated in the infectious sporozoite gene 4 (UIS4), but ETRAMP10.3 is understood to be an ortholog of UIS4 based on synteny and structural similarity. However, ETRAMP10.3 is not a functional ortholog of UIS4 and may have a different biological role. The biological function of ETRAMP10.3 is not yet fully understood, but its localization to vesicular structures in host erythrocytes suggests a role in host-parasite interactions or remodeling of infected erythrocytes. ETRAMP10.3 appears to play an important role in the life cycle of Plasmodium. When ETRAMP10.3 is deleted, the deletion can result in impaired hepatic phase development and progression of the asexual hematopoietic phase in mice.

[0351] While the terms "UIS4" and "ETRAMP10.3" are sometimes used in the literature to refer to different proteins, in the context of this disclosure, the terms "UIS4" and "ETRAMP10.3" are used interchangeably to refer to ETRAMP10.3.

[0352] The plasmodium UIS4 sequence is publicly known (see, for example, UniProt accession number Q8IJM9, which is incorporated herein by reference in its entirety). An exemplary UIS4 amino acid sequence is provided in Table 2.

[0353] Liver-specific protein 1 (LISP-1) Liver-specific protein 1 (LISP-1) is expressed during plasmodium development in hepatocytes and localizes to the parasitic spore membrane (PVM) (see, e.g., Ishino et al., Cell Microbiol. 2009 Sep;11(9):1329-1339, the entire paper of which is incorporated herein by reference). LISP-1 is expressed at high levels during late hepatic development and has been shown to be involved in PVM disruption and subsequent merozoite release (see, e.g., Ishino et al., Cell Microbiol. 2009 Sep;11(9):1329-1339, the entire paper of which is incorporated herein by reference).

[0354] Intracellular plasmodium lacking LISP-1 develops in hepatic merozoites and exhibits normal infectivity to erythrocytes (see, e.g., Ishino et al., Cell Microbiol. 2009 Sep;11(9):1329-1339, the entire article of which is incorporated herein by reference). However, LISP1-deficient hepatic plasmodium does not rupture PVMs and remains trapped within hepatocytes (see, e.g., Ishino et al., Cell Microbiol. 2009 Sep;11(9):1329-1339, the entire article of which is incorporated herein by reference).

[0355] The plasmodium LISP-1 sequence is publicly known (see, for example, UniProt accession numbers A0A2I0C2X6, Q8ILR5, each of which is incorporated herein by reference in its entirety). Exemplary LISP-1 amino acid sequences are provided in Table 2.

[0356] Liver-specific protein 2 (LISP-2) Liver-specific protein 2 (LISP-2) contains a modified 6-cys domain and is expressed during plasmodium development in hepatocytes (see, e.g., Orito et al., Mol Microbiol. 2013 Jan;87(1):66-79, the entire report of which is incorporated herein by reference). LISP-2 has been shown to be expressed by hepatic-phase plasmodium, secreted into hepatocytes, and distributed throughout the host cell, including the nucleus (see, e.g., Orito et al., Mol Microbiol. 2013 Jan;87(1):66-79, the entire report of which is incorporated herein by reference).

[0357] Intracellular plasmodium lacking LISP2 does not mature effectively during merozoite development (see, for example, Orito et al., Mol Microbiol. 2013 Jan;87(1):66-79, which is incorporated herein by reference in its entirety).

[0358] Plasmodium LISP-2 sequences are publicly known (see, for example, UniProt accession numbers A0A2I0BZR4, Q8I1X6, Q9U0D4, each of which is incorporated herein by reference in its entirety). Exemplary LISP-2 amino acid sequences are provided in Table 2.

[0359] Thrombospongin-related adhesion protein (TRAP) Thrombospondin-associated adhesion proteins (TRAPs) contain an N-terminal domain commonly referred to as the von Willebrand factor A domain, but they also contain the binding magnesium necessary for sporozoite motility in vitro and infection in vivo. 2+It is most similar to an integrin I domain because it contains a metal ion-dependent adhesion site (MIDAS) containing ions (see, for example, Lu et al., PLoS One. 2020;15(1):e0216260, which is incorporated herein by reference in its entirety). The I domain is inserted into an extensible β ribbon, followed by a thrombospongin repeat (TSR) domain, a C-terminal proline-rich segment, a single-pass transmembrane domain, and a cytoplasmic domain (see, for example, Lu et al., PLoS One. 2020;15(1):e0216260, the entire article of which is incorporated herein by reference). Sequence analysis of the proline-rich segment revealed the presence of an SH3 domain-binding PxxP motif in the plasmodium TRAP (see, for example, Akhouri et al., Malar J. 2008 Apr 22;7:63.doi:10.1186 / 1475-2875-7-63, the entire article of which is incorporated herein by reference).

[0360] When the parasite comes into contact with the host cell, the TRAP is stored within the microname and becomes surface-exposed at the anterior tip of the sporozoite (see Akhouri et al., Malar J. 2008 Apr 22;7:63.doi:10.1186 / 1475-2875-7-63, the whole of which is incorporated herein by reference). The TRAP also plays an important role in sporozoite entry into hepatocytes by assisting sporozoites in gliding motility and in the recognition of host receptors on mosquito salivary glands and hepatocytes (see Akhouri et al., Malar J. 2008 Apr 22;7:63.doi:10.1186 / 1475-2875-7-63, the whole of which is incorporated herein by reference).

[0361] Plasmodium TRAP sequences are publicly known (see, for example, UniProt accession numbers A0A5Q2EXK8, A0A5Q2EZD7, A0A5Q2F1F6, A0A5Q2F2B8, A0A5Q2F2H6, A0A5Q2F4G9, O76110, P16893, Q01507, Q26020, Q76NM2, W8VNB6, each of which is incorporated herein by reference in its entirety), and exemplary TRAP amino acid sequences are provided in Table 2.

[0362] Liver phase-related protein-1 (LSAP-1) Hepatic phase-associated protein (LSAP-1) is found primarily around intracellular hepatic parasites throughout development, but not in hematopoietic parasites, and is likely found in small amounts in salivary gland sporozoites (see, e.g., Siau et al., PLoS Pathog. 2008 Aug 8;4(8):e1000121, the entire work of which is incorporated herein by reference). Although LSAP-1 is one of the most abundant transcripts in the salivary gland transcriptome, it has not been detected in proteomic studies of sporozoites. Rather, its expression has been detected only in the hepatic phase (see, e.g., Siau et al., PLoS Pathog. 2008 Aug 8;4(8):e1000121, the entire work of which is incorporated herein by reference).

[0363] The plasmodium LSAP-1 sequence is publicly known (see, for example, UniProt accession numbers Q8I632, W7JR53, each of which is incorporated herein by reference in its entirety). Exemplary LSAP-1 amino acid sequences are provided in Table 2.

[0364] Liver phase-related protein-2 (LSAP-2) Like LSAP-1, LSAP-2 is one of the most abundant transcripts in the salivary gland transcriptome, but it has not been detected in sporozoite proteomic studies. LSAP-2 has shown some efficacy as a vaccine when combined with other antigens. See, for example, Halbroth et al., Infect Immun. 2020 Jan 22;88(2):e00573-19.doi:10.1128 / IAI.00573-19.Print 2020 Jan 22 (the entire work is incorporated herein by reference).

[0365] The plasmodium LSAP-2 sequence is publicly known (see, for example, UniProt accession numbers Q8I632, W7JR53, each of which is incorporated herein by reference in its entirety). Exemplary LSAP-2 amino acid sequences are provided in Table 2.

[0366] Liver phase antigen 1 (LSA-1) Hepatic phase antigen 1 (LSA-1) is expressed after plasmodium enters hepatocytes and the antigen accumulates within the parasitic vesicles (see, for example, Tucker, K. et al., 2016, "Pre-Erythrocytic Vaccine Candidates in Malaria," in AJRodriguez-Morales (ed.), Current Topics in Malaria, IntechOpen, London. 10.5772 / 65592, each of which is incorporated herein by reference in whole). The function of LSA-1 remains unknown (see, for example, Tucker, K. et al., 2016, "Pre-Erythrocytic Vaccine Candidates in Malaria," in AJRodriguez-Morales (ed.), Current Topics in Malaria, IntechOpen, London. 10.5772 / 65592, each of which is incorporated herein by reference in whole).

[0367] LSA-1 is a 230 kDa proerythrocyte-phase protein containing a large central region consisting of more than 80 17-amino acid repeat units adjacent to highly conserved C and N-terminal regions (Richie, TLand Parekh, FK (2009) Malaria, the entire text is incorporated herein by reference). (In Vaccines for Biodefense and Emerging and Neglected Diseases (Barrett, ADT and Stanberry LR, eds), pp. 1309-1364, Elsevier, the entire text is incorporated herein by reference). LSA1 is expressed only by hepatic-phase plasmodium, not by sporozoites (Richie, TLand Parekh, FK (2009) Malaria, the entire text is incorporated herein by reference). In Vaccines for Biodefense and Emerging and Neglected Diseases (Barrett, ADT and Stanberry LR, eds), pp. 1309-1364, Elsevier (the entire article is incorporated herein by reference). Repeated regions result in significant protein variability among strains of Plasmodium falciparum (see, for example, Tucker, K. et al., 2016, "Pre-Erythrocytic Vaccine Candidates in Malaria," in AJ Rodriguez-Morales (ed.), Current Topics in Malaria, IntechOpen, London. 10.5772 / 65592 (the entire article is incorporated herein by reference)).

[0368] The plasmodium LSA-1 sequence is publicly known (see, for example, UniProt accession numbers Q25886, Q25887, Q25893, Q26028, Q9GTX5, O96125, each of which is incorporated herein by reference in its entirety). Exemplary LSA-1 amino acid sequences are provided in Table 2.

[0369] Liver phase antigen 3 (LSA-3) Hepatic phase antigen 3 (LSA-3) is a 200-kDa protein composed of two short repeat regions and one long repeat region flanked by three non-repeat regions (NR-A, NR-B, and NR-C) (see, for example, Tucker, K. et al., 2016, "Pre-Erythrocytic Vaccine Candidates in Malaria", in AJ Rodriguez-Morales (ed.), the entire protein of which is incorporated herein by reference, and Current Topics in Malaria, IntechOpen, London. 10.5772 / 65592, the entire protein of which is incorporated herein by reference). Non-repeating regions are well conserved across geographically diverse strains of Plasmodium falciparum (see, e.g., Tucker, K. et al., 2016, "Pre-Erythrocytic Vaccine Candidates in Malaria," in AJRodriguez-Morales (ed.), Current Topics in Malaria, IntechOpen, London. 10.5772 / 65592, which is incorporated herein by reference in its entirety). The most notable variability is in the repeating regions, resulting not from the composition of the repeating regions themselves, but from the composition and number of repeating subunits (see, e.g., Tucker, K. et al., 2016, "Pre-Erythrocytic Vaccine Candidates in Malaria," in AJRodriguez-Morales (ed.), Current Topics in Malaria, IntechOpen, London. 10.5772 / 65592, which is incorporated herein by reference in its entirety).

[0370] Recent in vitro data have shown that antibodies against LSA-3 (particularly the C-terminal portion of LSA-3) may provide some protection (see, for example, Morita et al, Sci Rep. 2017 Apr 5;7:46086.doi:10.1038 / srep46086, the entire report of which is incorporated herein by reference).

[0371] Plasmodium LSA-3 sequences are publicly known (e.g., UniProt accession numbers C7DU21, C7DU22, C7DU23, C7DU24, C7DU25, C7DU26, C7DU27, C7DU28, C7DU29, C7DU32, C7DU33, C7DU34, C7DU36, C7DU37, C7DU38, C7DU39, C7DU40, Q8I042, Q8I0A5, Q8I0D0, Q8IFR1, Q8IFR2, Q8IFR3, Q8IFR4, Q8I See FR5, Q8IFR6, Q8IFR7, Q8IFR8, Q8IFR9, Q8IFS0, Q8IFS1, Q8IFS2, Q8IFS3, Q8IFS4, Q8IFS5, Q8IFS6, Q8IFS7, Q8IFS8, Q8IFS9, Q8IFT0, Q8IFT1, Q8IFT2, Q8IFT3, Q8IFT4, Q9U0N9, Q9U0P0, A0A2I0BVD6, A0PFM9, and O96275 (each of which is incorporated herein by reference in whole). Exemplary LSA-3 amino acid sequences are provided in Table 2.

[0372] Glutamate-rich protein (GARP) Glutamate-rich protein (GARP) is an 80 kDA protein whose name is derived from its glutamate-rich amino acid sequence, which accounts for 24% of all residues. GARP is primarily expressed in the cyclic and trophic phases and is not essential for cell culture, although it has been shown to be highly immunogenic in animal models. While GARP is not essential for cell culture, its localization of infected erythrocytes to the periphery may indicate a role in the sequestration of infected erythrocytes. It has been proposed that GARP's involvement in sequestration occurs through binding with chloride / bicarbonate anion exchangers. Antibodies against GARP have been proposed to serve as a signature for protection against severe malaria, and their efficacy has been demonstrated in experimental studies in monkeys (see, e.g., Hon et al, Trends in Paras 2020 Aug;36(8):653-655.doi:10.1016 / j.pt.2020.05.012 and Lau et al, Plos Path.2014 10,e1004135, each of which is incorporated herein by reference in its entirety). The GARP sequence is publicly known (see, e.g., UniProt accession numbers, Q9GTW3, Q9U0N1, each of which is incorporated herein by reference in its entirety), and exemplary GARP amino acid sequences are provided in Table 2.

[0373] Parasite-infected red blood cell-specific protein 2 (PIESP2) Parasite-specific erythrocyte-specific protein 2 (PIESP2) (see, e.g., UniProt accession number Q8I488) is a highly immunogenic protein that is first expressed during the trophoblast stage and is thought to be important for the clinical progression of cerebral malaria. This protein is mainly found within red blood cells, but has been shown to be present on the surface of red blood cells, enabling it to adhere to endothelial cells of the cerebral vascular system. Antibodies against PIESP2 have been shown to prevent plasmodium vascular adhesion and may prove useful in preventing inflammatory responses in the brain and blood-brain barrier damage during cerebral malaria progression (see, e.g., Liu et al, Int J Biol Macromol. 2021 Apr 30;177:535-547.doi:10.1016 / j.ijbiomac.2021.02.145, the entire paper of which is incorporated herein by reference). The PIESP2 sequence is publicly known (see, for example, UniProt accession number Q8I488, which is incorporated herein by reference in its entirety), and an exemplary PIESP2 amino acid sequence is provided in Table 2.

[0374] Shizont exit antigen-1 (SEA1) Shizont exit antigen-1 (SEA1) is a large 244 kDA protein lacking a transmembrane domain or known targeting signal. The function of SEA1 is unknown; however, it has been shown to be effective in rodent vaccine studies and has even been proposed as a target for protective antibodies found in children. SEA1 received its name after it was reported that antibodies against this protein inhibited plasmodium merozoite escape. SEA1 localizes close to the centromere during nuclear fission, suggesting its role in the replication process. To date, various studies have proposed not only a role for SEA1 in escape but also a role in nuclear mitosis during replication. (Perrin et al. 2021, the entire work is incorporated herein by reference) (see, for example, Perrin et al, mBio. 2021 Mar 9;12(2):e03377-20.doi:10.1128 / mBio.03377-20, the entire work is incorporated herein by reference). The SEA1 sequence is publicly known (see, for example, UniProt accession number A0A143ZXM2, the entire work is incorporated herein by reference), and an exemplary SEA1 amino acid sequence is provided in Table 2.

[0375] [Table 3]

[0376] [Table 4]

[0377] [Table 5]

[0378] [Table 6]

[0379] [Table 7]

[0380] [Table 8]

[0381] [Table 9]

[0382] II. Plasmodium polypeptide constructs This disclosure, in particular, utilizes RNA technology as a modality for expressing one or more plasmodium polypeptide constructs comprising one or more malaria proteins or one or more portions thereof as described herein. For example, in some embodiments, the plasmodium polypeptide construct comprises one or more plasmodium Rh5 entry complex polypeptides or portions thereof (e.g., antigenic portions of one or more plasmodium Rh5 entry complex polypeptides). In some embodiments, one or more polypeptides or portions of the plasmodium Rh5 entry complex may comprise one or more polypeptides or portions of plasmodium reticulocyte-binding protein homolog 5 (Rh5), plasmodium cysteine-rich protective antigen (CyRPA), plasmodium Rh5 interacting protein (Ripr), plasmodium P113, plasmodium thrombospondin-associated apical merozoite protein (TRAMP), and / or plasmodium cysteine-rich microsecretory protein (CSS). In some embodiments, the portion of the Rh5 entry complex polypeptide may be a characteristic portion of the Rh5 entry complex polypeptide. In some embodiments, the plasmodium polypeptide construct further comprises one or more further amino acid sequences, such as a secretory signal (e.g., a heterologous secretory signal), a transmembrane region (e.g., a heterologous transmembrane region), a polymerizing region, and / or a linker, as described herein.

[0383] A. Specific plasmodium Rh5 entry complex polypeptide or its portion 1. Rh5 In some embodiments, the plasmodium polypeptide constructs described herein include one or more plasmodium Rh5 intrusion complex polypeptides or portions thereof (e.g., the antigenic moiety) comprising one or more plasmodium reticulocyte-binding protein homolog 5 (Rh5) polypeptides or their antigenic moieties.

[0384] In some embodiments, the plasmodium polypeptide constructs described herein include one or more polypeptides or antigenic moieties of a Rh5 polypeptide, e.g., Plasmodium Rh5 polypeptide, e.g., Plasmodium falciparum Rh5 (SEQ ID NO: 1) or a variant thereof (e.g., a glycosylated variant). In some embodiments, the Rh5 polypeptide may include a plasmodium Rh5 N-terminal disordered domain, a first plasmodium Rh5 ordered domain, a plasmodium Rh5 linked disordered domain, a second plasmodium Rh5 ordered domain (e.g., a C-terminal plasmodium Rh5 ordered domain), or a combination thereof. A portion of Rh5 (or a Rh5 polypeptide moiety) (e.g., an antigenic moiety of Rh5 or a Rh5 antigenic moiety) may refer to a portion of a Rh5 polypeptide domain or a portion spanning two or more Rh5 polypeptide domains.

[0385] In some embodiments, the plasmodium polypeptide construct comprises one or more antigenic moieties of Rh5. In some embodiments, one or more antigenic moieties of Rh5 comprises one or more Rh5 ordered domains. In some embodiments, the plasmodium Rh5 polypeptide or its antigenic moiety comprises 25, 30, 35, 40, or 45 consecutive amino acids of the Rh5 ordered domain or a variant thereof (e.g., a glycosylated variant).

[0386] In some embodiments, the plasmodium Rh5 antigenic moiety includes an Rh5 ordered domain corresponding to amino acids 140-247 or amino acids 146-247 of wild-type Rh5 (SEQ ID NO: 1) or its variant (e.g., a glycosylated variant). In some embodiments, the plasmodium Rh5 antigenic moiety includes an Rh5 ordered domain corresponding to amino acids 297-526 of wild-type Rh5 sequence (SEQ ID NO: 1) or its variant (e.g., a glycosylated variant). In some embodiments, one or more antigenic moieties of Rh5 include (i) an Rh5 ordered domain corresponding to amino acids 140-247 or amino acids 146-247 of wild-type Rh5 (SEQ ID NO: 1) or its variant, and / or (ii) an Rh5 ordered domain corresponding to amino acids 297-526 of wild-type Rh5 sequence (SEQ ID NO: 1) or its variant.

[0387] Rh5 ordered domains 140-247 (SEQ ID NO: 118) KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKINETYDKVKSKCNDIKNDLIATIKKLEHPYDINNK

[0388] Rh5 ΔPMX ordered domains 146-247 (SEQ ID NO: 119) QYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKINETYDKVKSKCNDIKNDLIATIKKLEHPYDINNK

[0389] Rh5 ordered domains 297-526 (SEQ ID NO: 120) NRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDMTNILQQSELLLTNLNKKMGSYIYIDTIK FIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDHLRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0390] In some embodiments, one or more plasmodium Rh5 antigenic moieties contain one or more unpaired cysteines. In some embodiments, one or more plasmodium Rh5 antigenic moieties contain two unpaired cysteines. In some embodiments, one or more plasmodium Rh5 antigenic moieties contain cysteine ​​at position 203, position 329, or both, as numbered according to SEQ ID NO: 1. In some embodiments, one or more plasmodium Rh5 antigenic moieties contain tyrosine at position 203, position 329, or both, as numbered according to SEQ ID NO: 1.

[0391] Rh5-C203Y ordered domains 140-247 (SEQ ID NO: 121) KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKINETYDKVKSKCNDIKNDLIATIKKLEHPYDINNK

[0392] Rh5 ΔPMX-C203Y ordered domains 146-247 (SEQ ID NO: 122) QYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKINETYDKVKSKCNDIKNDLIATIKKLEHPYDINNK

[0393] Rh5-C329Y ordered domains 297-526 (sequence number 123) NRTFKKMMDEYNTKKKKLIKCIKNHENDFNKIYMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDMTNILQQSELLLTNLNKKMGSYIYIDTIK FIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDHLRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0394] In some embodiments, one or more plasmodium Rh5 antigenic moieties include one, two, three, or four N-linked glycosylation sites. In some embodiments, one or more plasmodium Rh5 antigenic moieties include two N-linked glycosylation sites. In some embodiments, one or more plasmodium Rh5 antigenic moieties include one or more amino acid substitutions at one or more N-linked glycosylation sites, where the amino acid substitution prevents glycosylation. In some embodiments, the amino acid substitution preventing glycosylation includes a substitution from NX[T / S] to QX[T / S]. In some embodiments, the amino acid substitution preventing glycosylation includes a substitution from NX[T / S] to NXA. In some embodiments, one or more plasmodium Rh5 antigenic moieties include amino acid substitutions at all N-linked glycosylation sites, where the amino acid substitution prevents glycosylation.

[0395] In some embodiments, the plasmodium Rh5 antigenic moiety includes an Rh5 ordered domain corresponding to amino acids 140-247 of wild-type Rh5 (SEQ ID NO: 1) (or amino acids 140-247 of SEQ ID NO: 1 with 1, 2, 3, 4, or 5 amino acid substitutions). In some embodiments, the Rh5 ordered domain corresponds to amino acids 140-247 of wild-type Rh5 (SEQ ID NO: 1) and contains cysteine ​​at position 203. In some embodiments, the Rh5 ordered domain corresponds to amino acids 140-247 of wild-type Rh5 (SEQ ID NO: 1) and contains tyrosine at position 203. In some embodiments, the Rh5 ordered domain corresponds to amino acids 140-247 of wild-type Rh5 (SEQ ID NO: 1) and contains an N-linked glycosylation site at N214. In some embodiments, the Rh5 ordered domain corresponds to amino acids 140-247 of wild-type Rh5 (SEQ ID NO: 1) and contains an amino acid substitution that prevents glycosylation at position 214. In some embodiments, the amino acid substitution that prevents glycosylation at position 214 includes a substitution from NX[T / S] to QX[T / S]. In some embodiments, the amino acid substitution that prevents glycosylation at position 214 includes a substitution from NX[T / S] to NXA.

[0396] Rh5-N214Q ordered domains 140-247 (SEQ ID NO: 124) KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKIQETYDKVKSKCNDIKNDLIATIKKLEHPYDINNK

[0397] In some embodiments, the Rh5 ordered domain corresponds to amino acids 140-247 of wild-type Rh5 (SEQ ID NO: 1), and contains tyrosine at position 203 and glutamine at position 214.

[0398] Ordered domains 140-247 of Rh5-C203Y-N214Q (SEQ ID NO: 125) KNVNFLQYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKIQETYDKVKSKCNDIKNDLIATIKKLEHPYDINNK

[0399] In some embodiments, the Rh5 antigenic moiety (e.g., the Rh5 ordered domain) includes a plasmomepsin X (PMX) cleavage site. PMX is a conserved aspartate protease that is important for the proteolytic treatment and activation of proteins and other proteases involved in the progression and / or development of the plasmodium life cycle.

[0400] In some embodiments, the PMX cleavage site of the Rh5 ordered domain includes the sequence FLQY (SEQ ID NO: 36), and the cleavage occurs between L and Q. In some embodiments, the cleaved Rh5 ordered domain corresponds to amino acids 146-247 of wild-type Rh5 (SEQ ID NO: 1) or its variant. In some embodiments, the Rh5 antigenic moiety (e.g., the Rh5 ordered domain) does not contain a plasmmepsin X (PMX) cleavage site.

[0401] In some embodiments, the plasmodium Rh5 antigenic moiety includes an Rh5 ordered domain corresponding to amino acids 146-247 of wild-type Rh5 (SEQ ID NO: 1) (or amino acids 146-247 of SEQ ID NO: 1 with 1, 2, 3, 4, or 5 amino acid substitutions). In some embodiments, the Rh5 ordered domain corresponds to amino acids 146-247 of wild-type Rh5 (SEQ ID NO: 1) and contains cysteine ​​at position 203. In some embodiments, the Rh5 ordered domain corresponds to amino acids 146-247 of wild-type Rh5 (SEQ ID NO: 1) and contains tyrosine at position 203. In some embodiments, the Rh5 ordered domain corresponds to amino acids 146-247 of wild-type Rh5 (SEQ ID NO: 1) and contains an N-linked glycosylation site at N214. In some embodiments, the Rh5 ordered domain corresponds to amino acids 146-247 of wild-type Rh5 (SEQ ID NO: 1) and contains an amino acid substitution that prevents glycosylation at position 214. In some embodiments, the amino acid substitution that prevents glycosylation at position 214 includes a substitution from NX[T / S] to QX[T / S]. In some embodiments, the amino acid substitution that prevents glycosylation at position 214 includes a substitution from NX[T / S] to NXA.

[0402] Rh5 ΔPMX-N214Q ordered domains 146-247 (sequence number 126) QYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKCIAVDAFIKKIQETYDKVKSKCNDIKNDLIATIKKLEHPYDINNK

[0403] In some embodiments, the Rh5 ordered domain corresponds to amino acids 146-247 of wild-type Rh5 (SEQ ID NO: 1), and contains tyrosine at position 203 and glutamine at position 214.

[0404] Rh5 ΔPMX-C203Y-N214Q ordered domains 146-247 (sequence number 127) QYHFKELSNYNIANSIDILQEKEGHLDFVIIPHYTFLDYYKHLSYNSIYHKSSTYGKYIAVDAFIKKIQETYDKVKSKCNDIKNDLIATIKKLEHPYDINNK

[0405] In some embodiments, the plasmodium Rh5 antigenic moiety includes an Rh5 ordered domain corresponding to amino acids 297-526 of wild-type Rh5 (SEQ ID NO: 1) (or amino acids 297-526 of SEQ ID NO: 1 with 1, 2, 3, 4, or 5 amino acid substitutions). In some embodiments, the Rh5 ordered domain corresponds to amino acids 297-526 of wild-type Rh5 (SEQ ID NO: 1) and contains cysteine ​​at position 329. In some embodiments, the Rh5 ordered domain corresponds to amino acids 297-526 of wild-type Rh5 (SEQ ID NO: 1) and contains tyrosine at position 329.

[0406] In some embodiments, the Rh5 ordered domain corresponds to amino acids 297-526 of wild-type Rh5 (SEQ ID NO: 1) and includes an N-linked glycosylation site at N297. In some embodiments, the Rh5 ordered domain corresponds to amino acids 297-526 of wild-type Rh5 (SEQ ID NO: 1) and includes an amino acid substitution that prevents glycosylation at position 297. In some embodiments, the amino acid substitution that prevents glycosylation at position 297 includes a substitution from NX[T / S] to QX[T / S]. In some embodiments, the amino acid substitution that prevents glycosylation at position 297 includes a substitution from NX[T / S] to NXA.

[0407] Rh5 N297Q's ordered domains 297-526 (SEQ ID NO: 128) QRTFKKMMDEYNTKKKKLIKCIKNHENDFNKICMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDMTNILQQSELLLTNLNKKMGSYIYIDTIK FIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDHLRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0408] In some embodiments, the Rh5 ordered domain corresponds to amino acids 297-526 of wild-type Rh5 (SEQ ID NO: 1), and contains tyrosine at position 329 and / or glutamine at position 297.

[0409] In some embodiments, the Rh5 ordered domain corresponds to amino acids 297-526 of wild-type Rh5 (SEQ ID NO: 1), and contains cysteine ​​at position 329 and asparagine at position 297.

[0410] Rh5-N297Q-C329Y ordered domains 297-526 (sequence number 129) QRTFKKMMDEYNTKKKKLIKCIKNHENDFNKIYMDMKNYGTNLFEQLSCYNNNFCNTNGIRYHYDEYIHKLILSVKSKNLNKDLSDMTNILQQSELLLTNLNKKMGSYIYIDTIK FIHKEMKHIFNRIEYHTKIINDKTKIIQDKIKLNIWRTFQKDELLKRILDMSNEYSLFITSDHLRQMLYNTFYSKEKHLNNIFHHLIYVLQMKFNDVPIKMEYFQTYKKNKPLTQ

[0411] In some embodiments, the plasmodium Rh5 polypeptide or its antigenic moiety includes a Rh5 disordered domain or its antigenic moiety. In some embodiments, the Rh5 disordered domain is the Rh5 N-terminal disordered domain corresponding to amino acids 26-139 of wild-type Rh5 (SEQ ID NO: 1). In some embodiments, the Rh5 disordered domain is a Rh5 ligated disordered domain corresponding to amino acids 248-296 of wild-type Rh5 (SEQ ID NO: 1).

[0412] Rh5 disorder domains 26-139 (SEQ ID NO: 130) ENAIKKTKNQENNLTLLPIKSTEEEKDDIKNGKDIKKEIDNDKENIKTNNAKDHSTYIKSYLNTNVNDGLKYLFIPSHNSFIKKYSVFNQINDGMLLNEKNDVKNNEDYKNVDY

[0413] Rh5 disorder domains 248-296 (SEQ ID NO: 131) NDDSYRYDISEEIDDKSEETDDETEEVEDSIQDTDSNHTPSNKKKNDLM

[0414] In some embodiments, one or more plasmodium Rh5 antigenic moieties include two Rh5 ordered domains and one Rh5 disordered domain. In some embodiments, one or more plasmodium Rh5 antigenic moieties correspond to amino acids 140-526 of wild-type Rh5 (SEQ ID NO: 1). In some embodiments, one or more plasmodium Rh5 antigenic moieties correspond to amino acids 26-247 and 297-526 of wild-type Rh5 (SEQ ID NO: 1).

[0415] In some embodiments, the plasmodium polypeptide construct does not contain the Rh5 disordered domain. In some embodiments, the plasmodium Rh5 polypeptide or its antigenic moiety does not contain the Rh5 disordered domain corresponding to amino acids 26-139 or amino acids 248-296 of wild-type Rh5 (SEQ ID NO: 1). In some embodiments, the plasmodium polypeptide construct does not contain the Rh5 secretion signal, and does not include, for example, the sequence corresponding to amino acids 1-25 of wild-type Rh5 (SEQ ID NO: 1).

[0416] Rh5 secretion signal (SEQ ID NO: 132) MIRIKKKLILTIIYIHLFILNRLSF

[0417] In some embodiments, the plasmodium polypeptide construct comprises one or more plasmodium Rh5 polypeptides or their antigenic moieties, each containing two Rh5 ordered domains. In some embodiments, the plasmodium polypeptide construct comprises two plasmodium Rh5 antigenic moieties, each antigenic moiety containing an Rh5 ordered domain. In some embodiments, the plasmodium polypeptide construct comprises two Rh5 ordered domains corresponding to (i) amino acids 140-247 or 146-247 of wild-type Rh5 (SEQ ID NO: 1) or its variant, and (ii) amino acids 297-526 of wild-type Rh5 (SEQ ID NO: 1) or its variant. In some embodiments, the plasmodium polypeptide construct comprises two Rh5 ordered domains directly adjacent to each other.

[0418] In some embodiments, the plasmodium polypeptide construct comprises a Rh5 polypeptide containing exactly two antigenic moieties of Rh5, wherein the two antigenic moieties contain or consist of two Rh5 ordered domains. In some embodiments, the two Rh5 ordered domains correspond to (i) amino acids 140-247 or amino acids 146-247 of wild-type Rh5 (SEQ ID NO: 1) or its variant, and (ii) amino acids 297-526 of wild-type Rh5 (SEQ ID NO: 1) or its variant.

[0419] In some embodiments, the plasmodium polypeptide construct comprises two Rh5 ordered domains, the first Rh5 ordered domain corresponding to amino acids 140-247 or 146-247 of wild-type Rh5 (SEQ ID NO: 1), containing tyrosine at position 203 and glutamine at position 214, and the second Rh5 ordered domain corresponding to amino acids 297-526 of wild-type Rh5 (SEQ ID NO: 1), containing tyrosine at position 329 and / or glutamine at position 297.

[0420] In some embodiments, the plasmodium polypeptide construct comprises two Rh5 ordered domains, the first Rh5 ordered domain corresponding to amino acids 140-247 or 146-247 of wild-type Rh5 (SEQ ID NO: 1), containing tyrosine at position 203 and glutamine at position 214, and the second Rh5 ordered domain corresponding to amino acids 297-526 of wild-type Rh5 (SEQ ID NO: 1), containing cysteine ​​at position 329 and asparagine at position 297.

[0421] In some embodiments, the plasmodium polypeptide construct does not contain the Rh5 disorder domain (e.g., does not contain amino acids 26-139 or 248-296 of wild-type Rh5 (SEQ ID NO: 1)). In some embodiments, the plasmodium polypeptide construct does not contain the Rh5 secretion signal and does not contain the sequence corresponding to amino acids 1-25 of wild-type Rh5 (SEQ ID NO: 1).

[0422] In some embodiments, the plasmodium polypeptide construct comprises one or more antigenic moieties of plasmodium Rh5 containing a PMX cleavage site. In some embodiments, the plasmodium polypeptide construct comprises one or more antigenic moieties of plasmodium Rh5 containing or comprising the amino acid sequence of NFLQ (SEQ ID NO: 189) or a PMX cleavage site thereof.

[0423] 2. CyRPA In some embodiments, the plasmodium polypeptide constructs described herein include one or more plasmodium Rh5 intrusion complex polypeptides or portions thereof (e.g., the antigenic moiety) comprising one or more plasmodium cysteine-rich protective antigen (CyRPA) polypeptides or their antigenic moieties.

[0424] In some embodiments, the plasmodium polypeptide constructs described herein include one or more regions or parts (e.g., antigenic parts) of CyRPA, e.g., Plasmodium CyRPA, e.g., Plasmodium falciparum CyRPA (SEQ ID NO: 3) or its variants (e.g., one or more antigenic parts of CyRPA, e.g., Plasmodium CyRPA, e.g., Plasmodium falciparum CyRPA).

[0425] In some embodiments, the plasmodium polypeptide construct includes an antigenic moiety of CyRPA. In some embodiments, the plasmodium CyRPA polypeptide or its antigenic moiety includes 25, 30, 35, 40, or 45 consecutive amino acids of CyRPA or its variant (e.g., a glycosylated variant).

[0426] In some embodiments, the plasmodium CyRPA antigenic moiety does not include an endogenous signal sequence. In some embodiments, the plasmodium CyRPA antigenic moiety is operably linked to a heterologous signal sequence, as further described herein. In some embodiments, the plasmodium CyRPA antigenic moiety comprises or consists of amino acids 30-362 of wild-type CyRPA (SEQ ID NO: 3) or a variant thereof (e.g., a glycosylated variant). In some embodiments, the plasmodium CyRPA antigenic moiety comprises or consists of the sequence according to SEQ ID NO: 133 or a variant thereof (e.g., a glycosylated variant). In some embodiments, the plasmodium CyRPA antigenic moiety comprises or consists of a sequence having at least 85% sequence identity with SEQ ID NO: 133 or a variant thereof (e.g., a glycosylated variant).

[0427] CyRPA's 30-362 portion (sequence number 133) SRHVFIRTELSFIKNNVPCIRDMFFIYKRELYNICLDDLKGEEDETHIYVQKKVKDSWITLNDLFKETDLTGRPHIFAYVDVEEIIILLCEDEEFSNRKKDMTCHRFYSNDGKEYNNSEITISDYILKDKLLSSYVSLPLKIENREYFLICGVSPYKFKDDNKKDD ILCMASHDKGETWGTKIVIKYDNYKLGVQYFFLRPYISKNDLSFHFYVGDNINNVKNVNFIECTHEKDLEFVCSNRDFLKDNKVLQDVSTLNDEYIVSYGNDNNFAECYIFFNNENSILIKPEKYGNTTAGCYGGTFVKIDENRTLFIYSSSQGIYNIHTIYYANYE

[0428] In some embodiments, the plasmodium CyRPA antigenic moiety comprises or consists of amino acids 29-362 of wild-type CyRPA (SEQ ID NO: 3) or a variant thereof (e.g., a glycosylated variant). In some embodiments, the plasmodium CyRPA antigenic moiety comprises or consists of the sequence of SEQ ID NO: 169 or a variant thereof (e.g., a glycosylated variant). In some embodiments, the plasmodium CyRPA antigenic moiety comprises or consists of a sequence having at least 85% sequence identity with SEQ ID NO: 169 or a variant thereof (e.g., a glycosylated variant).

[0429] CyRPA's part 29-362 (sequence number 169) DSRHVFIRTELSFIKNNVPCIRDMFFIYKRELYNICLDDLKGEEDETHIYVQKKVKDSWITLNDLFKETDLTGRPHIFAYVDVEEIIILLCEDEEFSNRKKDMTCHRFYSNDGKEYNNSEITISDYILKDKLLSSYVSLPLKIENREYFLICGVSPYKFKDDNKKDD ILCMASHDKGETWGTKIVIKYDNYKLGVQYFFLRPYISKNDLSFHFYVGDNINNVKNVNFIECTHEKDLEFVCSNRDFLKDNKVLQDVSTLNDEYIVSYGNDNNFAECYIFFNNENSILIKPEKYGNTTAGCYGGTFVKIDENRTLFIYSSSQGIYNIHTIYYANYE

[0430] In some embodiments, one or more plasmodium CyRPA antigenic moieties include one or more N-linked glycosylation sites. In some embodiments, the plasmodium CyRPA antigenic moieties include one, two, or three N-linked glycosylation sites. In some embodiments, the plasmodium CyRPA antigenic moieties include amino acid substitutions at one or more N-linked glycosylation sites, where the amino acid substitutions prevent glycosylation. In some embodiments, the amino acid substitutions that prevent glycosylation include substitutions from NX[T / S] to QX[T / S]. In some embodiments, the amino acid substitutions that prevent glycosylation include substitutions from NX[T / S] to NXA. In some embodiments, the plasmodium CyRPA antigenic moieties include two amino acid substitutions at the N-linked glycosylation sites that prevent glycosylation. In some embodiments, the plasmodium CyRPA antigenic moieties include amino acid substitutions at all N-linked glycosylation sites, where the amino acid substitutions prevent glycosylation.

[0431] In some embodiments, the plasmodium CyRPA antigenic moiety contains asparagine at positions 145, 322, 338, or a combination thereof, as numbered according to SEQ ID NO: 133. In some embodiments, the plasmodium CyRPA antigenic moiety contains glutamine at positions 145, 322, 338, or a combination thereof, as numbered according to SEQ ID NO: 3. In some embodiments, the plasmodium CyRPA antigenic moiety contains glutamine at positions 145, 322, and 338, as numbered according to SEQ ID NO: 3. In some embodiments, the plasmodium CyRPA antigenic moiety contains or consists of the amino acid sequence of SEQ ID NO: 134.

[0432] In some embodiments, the plasmodium CyRPA antigenic moiety contains asparagine at positions 146, 323, 339, or a combination thereof, as numbered according to SEQ ID NO: 3. In some embodiments, the plasmodium CyRPA antigenic moiety contains glutamine at positions 146, 323, 339, or a combination thereof, as numbered according to SEQ ID NO: 3. In some embodiments, the plasmodium CyRPA antigenic moiety contains glutamine at positions 146, 323, and 339, as numbered according to SEQ ID NO: 3. In some embodiments, the plasmodium CyRPA antigenic moiety contains or consists of the amino acid sequence of SEQ ID NO: 170.

[0433] CyRPA Δglycan segment 30-362 (SEQ ID NO: 134) SRHVFIRTELSFIKNNVPCIRDMFFIYKRELYNICLDDLKGEEDETHIYVQKKVKDSWITLNDLFKETDLTGRPHIFAYVDVEEIIILLCEDEEFSNRKKDMTCHRFYSNDGKEY QNSEITISDYILKDKLLSSYVSLPLKIENREYFLICGVSPYKFKDDNKKDDILCMASHDKGETWGTKIVIKYDNYKLGVQYFFLRPYISKNDLSFHFYVGDNINNVKNVNFIECTHEKDLEFVCSNRDFLKDNKVLQDVSTLNDEYIVSYGNDNNFAECYIFFNNENSILIKPEKYG QTT AGCYGGTFVKIDE QRT LFIYSSSQGIYNIHTIYYANYE

[0434] CyRPA Δglycan segment 29-362 (SEQ ID NO: 170) DSRHVFIRTELSFIKNNVPCIRDMFFIYKRELYNICLDDLKGEEDETHIYVQKKVKDSWITLNDLFKETDLTGRPHIFAYVDVEEIIILLCEDEEFSNRKKDMTCHRFYSNDGKEY QNS EITISDYILKDKLLSSYVSLPLKIENREYFLICGVSPYKFKDDNKKDDILCMASHDKGETWGTKIVIKYDNYKLGVQYFFLRPYISKNDLSFHFYVGDNINNVKNVNFIECTHEKDLEFVCSNRDFLKDNKVLQDVSTLNDEYIVSYGNDNNFAECYIFFNNENSILIKPEKYG QTT AGCYGGTFVKIDE QRT LFIYSSSQGIYNIHTIYYANYE

[0435] 3. P113 In some embodiments, the plasmodium polypeptide constructs described herein comprise one or more plasmodium Rh5 intrusion complex polypeptides or portions thereof (e.g., the antigenic moiety) that include one or more plasmodium P113 polypeptides or their antigenic moieties.

[0436] In some embodiments, the plasmodium polypeptide constructs described herein include one or more regions or parts (e.g., antigenic parts) of P113, e.g., Plasmodium p113, e.g., Plasmodium falciparum P113 (SEQ ID NO: 6) or its variants (e.g., one or more antigenic parts of P113, e.g., Plasmodium p113, e.g., Plasmodium falciparum P113).

[0437] In some embodiments, the plasmodium polypeptide construct comprises one or more antigenic moieties of P113. In some embodiments, the plasmodium P113 polypeptide or its antigenic moiety comprises 25, 30, 35, 40, or 45 consecutive amino acids of the P113 polypeptide or its variant (e.g., a glycosylated variant). In some embodiments, the plasmodium polypeptide construct comprises the antigenic moiety of P113.

[0438] In some embodiments, the plasmodium P113 antigenic moiety does not include an endogenous signal sequence. In some embodiments, the plasmodium P113 antigenic moiety is operably linked to a heterologous signal sequence, as further described herein. In some embodiments, the plasmodium P113 antigenic moiety does not include a GPI anchor site. In some embodiments, the plasmodium P113 antigenic moiety includes or comprises amino acids 23-939 of wild-type P113 (SEQ ID NO: 6) or a variant (e.g., a glycosylated variant). In some embodiments, the plasmodium P113 antigenic moiety includes or comprises the sequence according to SEQ ID NO: 135. In some embodiments, the plasmodium P113 antigenic moiety includes or comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence according to SEQ ID NO: 135.

[0439] Sections 23-939 on page 113 (sequence number 135) YVHNDVIKFGEENSLKCSQGNLYVLHCEVQCLNGNNEIIHKRCNDDIEKKCNGNNKCIYFFEYELRKKTQSFRNKNSIEISECVESEQNEVKTSTTCLLSNSFILDEAFIQYFFFIKNKNEEPVICKDGNINIKSALLHSPFCEIKLKDISEYIRKKCDNNKECLIDPLDVQKNLLNEEDPCYINNSYVSVNVVCNKEEIGDESTDSSSMEIQDSTSNEQDENVKGMSSSQEMNSNNDENKNQDNESDDVNNNNNNNNDQDEQGNDGDVTSSMNKNEDNKDLEHGSSNDVNNNTDTLVNNKENKEFVLKEKSSLTSKINKELAHRTALFNKLADNISLLLNKKYDSFEIKDVLEDRYNEMKRDANPDVYYIYLMDTLDIEKIEDINLEEVKMSLLASLKETMNKIDTIEKKIEEEFKNKYISLYNKVKTTMPLELFDLNEDLVLLYNDFFDNGMIS SDIFFKYNPSENIMDHQEMVKKGSITEDELRIVNDLEPLDNYRRKRITELRKILVEKLRILILYKNNLFNTQASCIKSYCYKNPLNLKTLEVLLKKNYYRLKENKDYDVVSSIIQHLDNVDANKKKKWLTHERILKKLQVLIAEGYKRINEKEKDIDRRMAVYNALYEKAQSYNLQKLFNDSNDFLKKYAIMGNSFDDGDEVFGSQSSNFNIFDSNNTDQNNEQEQPKQDDQLLNNNNNDDVLSESNNENKEKTSDATHKETQEKSDQEPSQNIQEDNSDEKHAENEENVEQIETDSNVSEEANDENKDNMQTTDEGTEELQQNDEDAESLTKENSKSEEQENEDSTDAEAIDKEEVETEEKGKDEQKKDEQKEQDEEDGEKENKHKSSETTNETVTDIEENKNEVKGEEHLQGSEQSIEASESSQKDETKETEDKEEYVNANDDESSEEDTTPNE

[0440] In some embodiments, the plasmodium P113 antigenic moiety includes one or more N-linked glycosylation sites. In some embodiments, the plasmodium P113 antigenic moiety includes 1, 2, 3, 4, 5, 6, 7, or 8 N-linked glycosylation sites. In some embodiments, the plasmodium P113 antigenic moiety includes an amino acid substitution at one or more N-linked glycosylation sites, the amino acid substitution preventing glycosylation. In some embodiments, the amino acid substitution preventing glycosylation includes a substitution from NX[T / S] to QX[T / S]. In some embodiments, the amino acid substitution preventing glycosylation includes a substitution from NX[T / S] to NXA. In some embodiments, the plasmodium P113 antigenic moiety includes 2, 3, 4, 5, 6, or 7 amino acid substitutions at the N-linked glycosylation site that prevent glycosylation. In some embodiments, the plasmodium P113 antigenic moiety includes amino acid substitutions in all N-linked glycosylation sites, and the amino acid substitutions prevent glycosylation.

[0441] In some embodiments, the plasmodium P113 antigenic moiety contains asparagine at positions 207, 268, 317, 360, 661, 697, 779, 876, or combinations thereof, as numbered according to SEQ ID NO: 6. In some embodiments, the plasmodium P113 antigenic moiety contains glutamine at positions 207, 268, 317, 360, 661, 697, 779, 876, or combinations thereof, as numbered according to SEQ ID NO: 6. In some embodiments, the plasmodium P113 antigenic moiety contains glutamine at positions 207, 268, 317, 360, 661, 697, 779, and 854, as numbered according to SEQ ID NO: 6. In some embodiments, the plasmodium P113 antigenic moiety contains or consists of the amino acid sequence of SEQ ID NO: 136.

[0442] P113 Δglycan segment 23-939 (Sequence ID 136) YVHNDVIKFGEENSLKCSQGNLYVLHCEVQCLNGNNEIIHKRCNDDIEKKCNGNNKCIYFFEYELRKKTQSFRNKNSIEISECVESEQNEVKTSTTCLLSNSFILDEAFIQYFFFIKNKNEEPVICKDGNINIKSALLHSPFCEIKLKDISEYIRKKCDNNKECLIDPLDVQKNLLNEEDPCYI QNS YVSVNVVCNKEEEIGDESTDSSSMEIQDSTSNEQDENVKGMSSSQEMNSNNDENKNQD QES DDDVNNNNNNDDQDEQGNDGDVTSSMNKNEDNKDLEHGSSNDVN QNT DTLVNNKENKEFVLKEKSSLTSKINKELAHRTALFNKLAD QIS LLLNKKYDSFEIKDVLEDRYNEMKRDANPDVYYIYLMDTLDIEKIEDINLEEVKMSLASLKETMNKIDTIEKKIEEFKNKKYISLYNKVKTTMPELFDLNEDLVLYNDFPFDNGMISSDIFFKYNPSENIMDHQEMVKKGSITEDELR IVNDLEPLDNYRRRKRITELRKILVEKLRILYEKNNLFNTQASCIKSYCYKNPLNLKTLEVLLKKNYYRLKENKDYDVVSSIIQHLDNVDANKKKKWLTHERILKKLQVLIAEGYKRINEKEKDIDRRMAVYNALYEKAQSYNLQKLF QDS NDFLKKYAIMGNSFDDDGEVFGSQSSNFNIFDS QNT DQNNEQEQPKQDDQLLNNNNDDVLSENNENKEKTSDDATHKETQEKSDQEPSQNIQEDNSDEKHAENEENVEQIETDS QVS EEANDENKDNMQTTDEGTEELQQNDEDAESLTKENSKSEEQENEDSTDAEAIDKEEVETEEKGKDEQKKDEQKEQDEEEDGEKENKHKSSETT QET VTDIEENKNEVKGEEHLQGSEQSIEASESSQKDETKETEDKEEYVNANDDESSEEDTTPNE

[0443] 4. Ripple In some embodiments, the plasmodium polypeptide constructs described herein comprise one or more plasmodium Rh5 intrusion complex polypeptides or portions thereof (e.g., the antigenic moiety) comprising one or more plasmodium Rh5 interacting protein (Ripr) polypeptides or their antigenic moieties.

[0444] In some embodiments, the plasmodium polypeptide constructs described herein include one or more regions or portions (e.g., antigenic portions) of Ripr, e.g., Plasmodium Ripr, e.g., Plasmodium falciparum Ripr (SEQ ID NO: 2) or its variants (e.g., one or more antigenic portions of Ripr, e.g., Plasmodium Ripr, e.g., Plasmodium falciparum Ripr). In some embodiments, the antigenic portion of the plasmodium Ripr polypeptide includes or comprises the amino acid sequence given by SEQ ID NO: 178. In some embodiments, the antigenic portion of the plasmodium Ripr polypeptide includes or comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence given by SEQ ID NO: 178. In some embodiments, the antigenic portion of the plasmodium Ripr polypeptide includes or comprises the amino acid sequence given by SEQ ID NO: 177. In some embodiments, the antigenic portion of the plasmodium Ripr polypeptide includes or comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence given by SEQ ID NO: 177. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide includes or consists of the amino acid sequence of SEQ ID NO: 176. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide includes or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 176. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide includes or consists of the amino acid sequence of SEQ ID NO: 175. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide includes or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 175. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide includes or consists of the amino acid sequence of SEQ ID NO: 174. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide includes or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 174.In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide includes or consists of the amino acid sequence of SEQ ID NO: 173. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide includes or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 173. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide includes or consists of the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide includes or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 172. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide includes or consists of the amino acid sequence of SEQ ID NO: 171. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide includes or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 171.

[0445] Ripr part 20-1086 (sequence number 171)

[0446] Ripr Δglycan segment 20-1086 (SEQ ID NO: 172)

[0447] Ripr Δglycan segment 530-1086 (SEQ ID NO: 173) LEIQNEISSHNSNQFSTDPHTNSNNINNMNIKKVEIFRSRFSSKLQCQGGKINIDKAILKGGEGCNDLLLTNSLKSYCNDLSECDIGLIYHFDTYCINDQYLFVSYSCSNLCNKCHQNSTCYGNRFNYDCFCDNPYISK YGNKLCERPNDCESVLCSQNQVCQILPNDKLICQCEEGYKNVKGKCVPDNKCDLSCPSNKVCVIENGKQTCKCSERFVLENGVCICANDYKMEDGINCIAKNKCKRKEYENICTNPNEMCAYNEETDIVKCECKEHYYR SSRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCLINEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLEDKCVHNKKCSENSICVNVMNKEPICVCTYNYYKKDGVCLIQNPCLKD NGGCSRNSECTFKYSKIQCTCKENYKNKDDSCVPNTNEYDESFTFQYNDDASIILGACGMIEFSYIYNQIIWKIQNSKESYVFYYDYPTAGNIEVQIKNEIFHTIIYLKKKIGNSVIYDDFQVDHQTCIYENVFYYYSNQN

[0448] Ripr part 720-934 (sequence number 174) DLSCPSNKVCVIENGKQTCKCSERFVLENGVCICANDYKMEDGINCIAKNKCKRKEYENICTNPNEMCAYNEETDIVKCECKEHYYRSSRGECILNDYCKDINCKENEEC SIVNFKPECVCKENLKKNNKGECIYENSCLINEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLEDKCVHNKKCSENSICVNVMNKEPICVCTYNYYKKDGVCLIQ

[0449] Ripr part 769-900 (sequence number 175) KNKCKRKEYENICTNPNEMCAYNEETDIVKCECKEHYYRSSRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCLINEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLED

[0450] Ripr part 769-856 (sequence number 176) KNKCKRKEYENICTNPNEMCAYNEETDIVKCECKEHYYRSSRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYE

[0451] Ripr's 817-900 portion (sequence number 177) DYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCLINEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLED

[0452] Ripr part 817-856 (sequence number 178) DYCKDINCKENEECSIIVNFKPECVCKENLKKNNKGECIYE

[0453] In some embodiments, the plasmodium polypeptide construct comprises one or more antigenic moieties of Ripr. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide comprises 25, 30, 35, 40, or 45 consecutive amino acids of the Ripr polypeptide or its variant (e.g., a glycosylated variant). In some embodiments, the plasmodium polypeptide construct comprises one or more antigenic moieties of Ripr.

[0454] In some embodiments, the plasmodium Ripr antigenic moiety includes one or more N-linked glycosylation sites. In some embodiments, the plasmodium Ripr antigenic moiety includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 N-linked glycosylation sites. In some embodiments, the plasmodium Ripr antigenic moiety includes amino acid substitutions at one or more N-linked glycosylation sites, where the amino acid substitutions prevent glycosylation. In some embodiments, the amino acid substitutions that prevent glycosylation include substitutions from NX[T / S] to QX[T / S]. In some embodiments, the amino acid substitutions that prevent glycosylation include substitutions from NX[T / S] to NXA. In some embodiments, the plasmodium Ripr antigenic moiety includes amino acid substitutions at all N-linked glycosylation sites, where the amino acid substitutions prevent glycosylation.

[0455] In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide contains asparagine at positions 646, 964, 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide contains glutamine at positions 646, 964, 1021, or a combination thereof, as numbered according to SEQ ID NO: 2. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide contains glutamine at positions 646, 964, and 1021, as numbered according to SEQ ID NO: 2. In some embodiments, the antigenic moiety of plasmodium Ripr contains or consists of the amino acid sequence according to SEQ ID NO: 173.

[0456] Ripr part 530-1086 (sequence number 190) LEIQNEISSHNSNQFSTDPHTNSNNINNMNIKKVEIFRSRFSSKLQCQGGKINIDKAILKGGEGCNDLLLTNSLKSYCNDLSECDIGLIYHFDTYCINDQYLFVSYSCSNLCNKCHNNSTCYGNRFNYDCFCDNPYISKYGNKLCERPNDCESVLCSQNQVCQILPNDKLICQCEEGYKNVKGKCVPDNKCDLSCPSNKVCVIENGKQTCKCSERFVLENGVCICANDYKMEDGINCIAKNKCKRKEYENICTNPNEMCAYNEETDIVKCECKEHYYRSSRGECILNDYCKDINCKENEECSIVNFKPECVCKENLKKNNKGECIYENSCLINEGNCPKDSKCIYREYKPHECVCNKQGHVAVNGKCVLEDKCVHNKKCSENSICVNVMNKEPICVCTYNYYKKDGVCLIQNPCLKDNGGCSRNSECTFKYSKINCTCKENYKNKDDSCVPNTNEYDESFTFQYNDDASIILGACGMIEFSYIYNQIIWKINNSKESYVFYYDYPTAGNIEVQIKNEIFHTIIYLKKKIGNSVIYDDFQVDHQTCIYENVFYYSNQN

[0457] In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide contains asparagine at positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, 1021, or combinations thereof, as numbered according to SEQ ID NO: 2. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide contains glutamine at positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, 1021, or combinations thereof, as numbered according to SEQ ID NO: 2. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide contains glutamine at positions 103, 144, 228, 303, 334, 480, 498, 506, 526, 646, 964, and 1021, numbered according to SEQ ID NO: 2. In some embodiments, the antigenic moiety of the plasmodium Ripr polypeptide contains or consists of SEQ ID NO: 172.

[0458] In some embodiments, the plasmodium Ripr antigenic moiety includes a PMX cleavage site. In some embodiments, the PMX cleavage site of the Ripr antigenic moiety includes the sequence GNISMLEIQNEE (SEQ ID NO: 37). In some embodiments, the plasmodium Ripr antigenic moiety does not include a PMX cleavage site. In some embodiments, the PMX cleavage site of the Ripr antigenic moiety includes or consists of the sequence SMLE (SEQ ID NO: 191).

[0459] 5. TRAMP In some embodiments, the plasmodium polypeptide constructs described herein include one or more plasmodium Rh5 entry complex polypeptides or portions thereof (e.g., the antigenic moiety) comprising one or more plasmodium thrombospondin-associated apical merozoite protein (TRAMP) polypeptides or their antigenic moieties.

[0460] In some embodiments, the plasmodium polypeptide constructs described herein include one or more regions or portions of TRAMP, e.g., plasmodium TRAMP, e.g., Plasmodium falciparum TRAMP (SEQ ID NO: 4) or its variants (e.g., one or more antigenic portions of TRAMP, e.g., plasmodium TRAMP, e.g., Plasmodium falciparum TRAMP). In some embodiments, the antigenic portion of the plasmodium TRAMP polypeptide includes or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 179. In some embodiments, the antigenic portion of the plasmodium TRAMP polypeptide includes or consists of the amino acid sequence of SEQ ID NO: 179.

[0461] TRAMP section 42-352 (sequence number 179) LQVYQNNGNFKMEILEPECTIQKNLPSEEEKEEFIDFDRKELLLHDFVVIKNSQFTDEKTLELYLSKEENNKSYLMLTFYLGDLKLMIGHNSPYEISLILNVATVNKNMSNCKNNSYNIVLLKTTDVFNTSDLEILEGPIQFSLGKSSGAFRINV TNFFLNNTWKAFTKDKISFLIKPEENCYTILENKINKPQLLIEKKTTFYSEWGEWSNCSMDCDHPDNVQIRERECIHPSGDCFKGDLKESRPCIIPLPPCNELFSHKDNSTFKILMIILPIVLVISIITILYHIFYKRKGAEKELYENVAGRYMYD

[0462] In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide comprises 25, 30, 35, 40, or 45 consecutive amino acids of the TRAMP polypeptide or a variant thereof (e.g., a glycosylated variant). In some embodiments, the plasmodium polypeptide construct comprises the antigenic moiety of TRAMP.

[0463] In some embodiments, the plasmodium TRAMP antigenic moiety includes one or more N-linked glycosylation sites. In some embodiments, the plasmodium TRAMP antigenic moiety includes 1, 2, 3, 4, 5, 6, 7, or 8 N-linked glycosylation sites. In some embodiments, the plasmodium TRAMP antigenic moiety includes amino acid substitutions at one or more N-linked glycosylation sites, where the amino acid substitutions prevent glycosylation. In some embodiments, the amino acid substitutions that prevent glycosylation include substitutions from NX[T / S] to QX[T / S]. In some embodiments, the amino acid substitutions that prevent glycosylation include substitutions from NX[T / S] to NXA. In some embodiments, the plasmodium TRAMP antigenic moiety includes amino acid substitutions at all N-linked glycosylation sites, where the amino acid substitutions prevent glycosylation.

[0464] In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide contains asparagine at positions 149, 195, 202, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide contains glutamine at positions 149, 195, 202, or a combination thereof, as numbered according to SEQ ID NO: 4. In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide contains glutamine at positions 149, 195, and 202, as numbered according to SEQ ID NO: 4. In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide contains or consists of the amino acid sequence according to SEQ ID NO: 181.

[0465] TRAMP Δglycan segment 42-352 (SEQ ID NO: 181) LQVYQNNGNFKMEILEPECTIQKNLPSEEEKEEFIDFDRKELLLHDFVVIKNSQFTDEKTLELYLSKEENNKSYLMLTFYLGDLKLMIGHNSPYEISLILNVATVNKQMSNCKNNSYNIVLLKTTDVFNTSDLEILEGPIQFSLGKSSGAFRIQV TNFFLQNTWKAFTKDKISFLIKPEENCYTILENKINKPQLLIEKKTTFYSEWGEWSNCSMDCDHPDNVQIRERECIHPSGDCFKGDLKESRPCIIPLPPCNELFSHKDNSTFKILMIILPIVLVISIITILYHIFYKRKGAEKELYENVAGRYMYD

[0466] In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide contains asparagine at positions 112, 149, 155, 170, 195, 202, 253, 305, or combinations thereof, as numbered according to SEQ ID NO: 4. In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide contains glutamine at positions 112, 149, 155, 170, 195, 202, 253, 305, or combinations thereof, as numbered according to SEQ ID NO: 4. In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide contains glutamine at positions 112, 149, 155, 170, 195, 202, 253, and 305, as numbered according to SEQ ID NO: 4. In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide contains or consists of the amino acid sequence according to SEQ ID NO: 180.

[0467] TRAMP Δglycan segment 42-352 (SEQ ID NO: 180) LQVYQNNGNFKMEILEPECTIQKNLPSEEEKEEFIDFDRKELLLHDFVVIKNSQFTDEKTLELYLSKEENQKSYLMLTFYLGDLKLMIGHNSPYEISLILNVATVNKQMSNCKQNSYNIVLLKTTDVFQTSDLEILEGPIQFSLGKSSGAFRIQV TNFFLQNTWKAFTKDKISFLIKPEENCYTILENKINKPQLLIEKKTTFYSEWGEWSQCSMDDHPDNVQIRERECIHPSGDCFKGDLKESRPCIIPLPPCNELFSHKDQSTFKILMIILPIVLVISITILYHIFYKRKGAEKELYENVAGRYMYD

[0468] In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide includes a PMX cleavage site. In some embodiments, the PMX cleavage site of the TRAMP antigenic moiety includes or consists of the sequence HFLQ (SEQ ID NO: 192). In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide includes a SUB2 cleavage site. In some embodiments, the SUB2 cleavage site of the TRAMP antigenic moiety includes or consists of ILMIILPIVLVISIITILYHIFY (SEQ ID NO: 193).

[0469] In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide includes a transmembrane region (also referred to as a "transmembrane domain"). In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide includes a transmembrane region at its C-terminus. In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide includes an HSV transmembrane region, e.g., an HSV-1 or HSV-2 transmembrane region. In some embodiments, the antigenic moiety of the plasmodium TRAMP polypeptide includes or comprises an HSV gD transmembrane region, e.g., a transmembrane region including or comprising the amino acid sequence of SEQ ID NO: 75. In some embodiments, the plasmodium TRAMP polypeptide does not include a transmembrane region.

[0470] 6. CSS In some embodiments, the plasmodium polypeptide constructs described herein include one or more plasmodium Rh5 intrusion complex polypeptides or portions thereof (e.g., the antigenic moiety) comprising one or more plasmodium cysteine-rich microsecretory protein (CSS) polypeptides or their antigenic moieties.

[0471] In some embodiments, the plasmodium polypeptide constructs described herein include one or more regions or parts of CSS, e.g., Plasmodium CSS, e.g., Plasmodium falciparum CSS (SEQ ID NO: 5 or 214), or variants thereof (e.g., one or more antigenic parts of CSS, e.g., Plasmodium CSS, e.g., Plasmodium falciparum CSS).

[0472] In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains a C30S mutation numbered according to SEQ ID NO: 5 or 214. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains serine at position 30, numbered according to SEQ ID NO: 5 or 214.

[0473] In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide includes or comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 194. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide includes or comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 195. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide includes or comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 182. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide includes or comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO: 182.

[0474] CSS section 21-152 (array element 194) QDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQYGEDIKIIANKEYEINCMNNSKVFSPLKDTFINNTNIKLYSPKLHFEIKDITHKGKNAALYYLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKK

[0475] CSS code, lines 153-290 (array number 195) HINEEYSIFFEEDVHVCDFSKGNLDITPSAGFYLKNSRNVSCIYRVIPNKLFLIKLPKLDIVTEKLLPSIVNCLSEFSFINFTLKHVQEGDNYISFNVIFGEFKKHFNLTCSLDLSDFQQEPSNLGKTANITFIFSK

[0476] CSS code sections 21-290 (array number 182) QDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQYGEDIKIIANKEYEINCMNNSKVFCPLKDTFINNTNIKLYSPKLHFEIKDITHKGKNAALYYLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKKHIN EEYSIFNEEEDVHVCDFSKGNLDITPSAGFYLKNSRNVSCIYRVIPNKLFLIKLPKLDIVTEKLLPSIVNCLSEFSFINFTLKHVQEGDNYISFNVIFGEFKKHFNLTCSLDLSDFQQEPCNLGKTANITFIFSK

[0477] In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide comprises 25, 30, 35, 40, or 45 consecutive amino acids of the CSS polypeptide or a variant thereof (e.g., a glycosylated variant). In some embodiments, the plasmodium polypeptide construct comprises one or more antigenic moieties of CSS.

[0478] In some embodiments, the plasmodium CSS antigenic moiety includes one or more N-linked glycosylation sites. In some embodiments, the plasmodium CSS antigenic moiety includes one, two, three, four, five, or six N-linked glycosylation sites. In some embodiments, the plasmodium CSS antigenic moiety includes amino acid substitutions at one or more N-linked glycosylation sites, where the amino acid substitutions prevent glycosylation. In some embodiments, the amino acid substitutions that prevent glycosylation include substitutions from NX[T / S] to QX[T / S]. In some embodiments, the amino acid substitutions that prevent glycosylation include substitutions from NX[T / S] to NXA. In some embodiments, the plasmodium CSS antigenic moiety includes amino acid substitutions at all N-linked glycosylation sites, where the amino acid substitutions prevent glycosylation.

[0479] In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains serine at position 80, numbered according to SEQ ID NO: 5 or 214.

[0480] In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains asparagine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains glutamine at position 74, position 88, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains glutamine at positions 74 and 88, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains or consists of the amino acid sequence according to SEQ ID NO: 184.

[0481] CSS Δglycan part 21-152 (sequence number 184) QDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQYGEDIKIIANKEYEINCMQNSKVFSPLKDTFIQNTNIKLYSPKLHFEIKDITHKGKNAALYYLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKK

[0482] In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains serine at position 276, numbered according to SEQ ID NO: 5 or 214. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains asparagine at positions 192, 234, 261, 283, or a combination thereof, numbered according to SEQ ID NO: 5 or 214. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains glutamine at positions 192, 234, 261, 283, or a combination thereof, numbered according to SEQ ID NO: 5 or 214. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains glutamine at positions 192, 234, 261, and 283, numbered according to SEQ ID NO: 5 or 214. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains or consists of the amino acid sequence according to SEQ ID NO: 185.

[0483] CSS Δglycan part 153-290 (sequence number 185) HINEEYSIFFEEDVHVCDFSKGNLDITPSAGFYLKNSRQVSCIYRVIPNKLFLIKLPKLDIVTEKLLPSIVNCLSEFSFIQFTLKHVQEGDNYISFNVIFGEFKKHFQLTCSLDLSDFQQEPSNLGKTAQITFIFSK

[0484] In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains asparagine at positions 74, 88, 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains glutamine at positions 74, 88, 192, 234, 261, 283, or a combination thereof, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains glutamine at positions 74, 88, 192, 234, 261, and 283, as numbered according to SEQ ID NO: 5 or 214. In some embodiments, the antigenic moiety of the plasmodium CSS polypeptide contains or consists of the amino acid sequence according to SEQ ID NO: 183.

[0485] CSS Δglycan part 21-290 (sequence number 183) QDEKSVKNISVCDFTDKLNFLPLEKTKILCELKPQYGEDIKIIANKEYEINCMQNSKVFCPLKDTFIQNTNIKLYSPKLHFEIKDITHKGKNAALYYLKIDEEASDIFFSCSIKPKQVSGLLEGEVRVNLKKHIN EEYSIFNEEEDVHVCDFSKGNLDITPSAGFYLKNSRQVSCIYRVIPNKLFLIKLPKLDIVTEKLLPSIVNCLSEFSFIQFTLKHVQEGDNYISFNVIFGEFKKHFQLTCSLDLSDFQQEPCNLGKTAQITFIFSK

[0486] B. Secretory signals In some embodiments, the plasmodium polypeptide constructs described herein include a secretory signal that functions in, for example, mammalian cells. In some embodiments, the secretory signal utilized is a heterologous secretory signal. In some embodiments, the heterologous secretory signal includes or consists of a non-human secretory signal. In some embodiments, the heterologous secretory signal includes or consists of a viral secretory signal. In some embodiments, the viral secretory signal includes or consists of an HSV secretory signal (e.g., an HSV-1 or HSV-2 secretory signal). In some embodiments, the HSV secretory signal includes or consists of an HSV glycoprotein D (gD) secretory signal.

[0487] In some embodiments, the secretion signal includes or consists of an Ebola virus secretion signal. In some embodiments, the Ebola virus secretion signal includes or consists of an Ebola virus spike glycoprotein (SGP) secretion signal.

[0488] In some embodiments, the secretory signal is characterized by a length of approximately 15–30 amino acids.

[0489] In many embodiments, the secretory signal is located at the N-terminus of the plasmodium polypeptide construct described herein. In some embodiments, the secretory signal preferably enables the secretion of the plasmodium polypeptide construct it associates with into a defined cellular compartment, preferably the cell surface, endoplasmic reticulum (ER), or endosomal-lysosome compartment.

[0490] In some embodiments, the secretion signal includes or consists of a plasmodium secretion signal. In some embodiments, the plasmodium secretion signal includes or consists of a plasmodium CSP secretion signal (e.g., derived from Plasmodium falciparum, e.g., Plasmodium falciparum isolate 3D7 (SEQ ID NO: 7)). In some embodiments, the plasmodium secretion signal includes a plasmodium Rh5 secretion signal. In some embodiments, the plasmodium secretion signal includes a plasmodium CyRPA secretion signal. In some embodiments, the plasmodium secretion signal includes or consists of the amino acid sequence according to SEQ ID NO: 187. In some embodiments, the plasmodium secretion signal includes a plasmodium P113 secretion signal. In some embodiments, the plasmodium secretion signal includes a plasmodium Ripr secretion signal. In some embodiments, the plasmodium secretion signal includes or consists of the amino acid sequence according to SEQ ID NO: 186. In some embodiments, the plasmodium secretion signal includes a plasmodium TRAMP secretion signal. In some embodiments, the plasmodium secretion signal includes a plasmodium CSS secretion signal. In some embodiments, the plasmodium secretion signal includes or consists of the amino acid sequence given by SEQ ID NO: 188.

[0491] In some embodiments, the secretion signal is selected from S1S2 secretion signals (aa1-19), immunoglobulin secretion signals (aa1-22), human SPARC secretion signal, human insulin isoform 1 secretion signal, human albumin secretion signal, and the like. Those skilled in the art will recognize other secretion signals, for example, those disclosed in International Publication No. 2017 / 081082, which is incorporated herein by reference in whole (e.g., SEQ ID NOs: 1-1115 and 1728, or fragmentary variants thereof). In some embodiments, the plasmodium polypeptide constructs described herein do not contain a secretion signal.

[0492] In some embodiments, the secretory signal is one of those listed in Table 3, or a secretory signal having a difference of 1, 2, 3, 4, or 5 amino acids thereto. In some embodiments, the signal sequence is selected from those included in Table 3 below and / or encoded by the sequences in Table 4 below. [Table 10]

[0493] [Table 11]

[0494] C. Transmembrane region In some embodiments, the plasmodium polypeptide constructs described herein include a transmembrane region (also referred to as a “transmembrane domain”). In some embodiments, the transmembrane region is located at the N-terminus of the plasmodium polypeptide construct. In some embodiments, the transmembrane region is located at the C-terminus of the plasmodium polypeptide construct. In some embodiments, the transmembrane region is not located at the N-terminus or C-terminus of the plasmodium polypeptide construct.

[0495] Transmembrane regions are known in the art and any of them can be used in the plasmodium polypeptide constructs described herein. In some embodiments, the transmembrane regions include or are those of the hemagglutinin (HA) of influenza virus, Env of HIV-1, the G protein of equine infectious anemia virus (EIAV), mouse leukemia virus (MLV), mouse mammary tumor virus, vesicular stomatitis virus (VSV), rabies virus, or seven-transmembrane domain receptors.

[0496] In some embodiments, the heterologous transmembrane region does not include a hemagglutin transmembrane region. In some embodiments, the heterologous transmembrane region includes or consists of a non-human transmembrane region. In some embodiments, the heterologous transmembrane region includes or consists of a viral transmembrane region. In some embodiments, the heterologous transmembrane region includes or consists of an HSV transmembrane region, e.g., an HSV-1 or HSV-2 transmembrane region. In some embodiments, the HSV transmembrane region includes or consists of an HSV gD transmembrane region, e.g., the amino acid sequence GLIAGAVGGSLLAALVICGIVYWMRRHTQKAPKRIRLPHIR (SEQ ID NO: 75).

[0497] In some embodiments, the heterologous transmembrane region includes or comprises a human transmembrane region. In some embodiments, the human transmembrane region includes or comprises a human phacosylphosphatidylinositol (hDAF-GPI) anchor region. In some embodiments, the hDAF-GPI anchor region includes or comprises the amino acid sequence PNKGSGTTSGTTRLLSGHTCFTLTGLLGTLVTMGLLT (SEQ ID NO: 76).

[0498] In some embodiments, the transmembrane region includes or comprises a plasmodium transmembrane region. In some embodiments, the transmembrane region utilized is typically associated with CSP in nature. In some embodiments, the plasmodium transmembrane region includes or comprises a plasmodium CSP glycosylphosphatidylinositol (GPI) anchor region. In some embodiments, the plasmodium CSP GPI anchor region is derived from Plasmodium falciparum. In some embodiments, the plasmodium CSP GPI anchor region is derived from Plasmodium falciparum isolate 3D7 (SEQ ID NO: 7), for example, amino acids 374-397 of SEQ ID NO: 7.

[0499] In some embodiments, the transmembrane region includes a plasmodium TRAP transmembrane region. In some embodiments, the transmembrane region includes a plasmodium P113 transmembrane region.

[0500] In some embodiments, the transmembrane region used is a heterogeneous transmembrane region.

[0501] In some embodiments, the plasmodium polypeptide constructs described herein do not include transmembrane regions.

[0502] Exemplary transmembrane structures are provided in Table 5 below:

[0503] [Table 12]

[0504] D. Multimerization region In some embodiments, the plasmodium polypeptide constructs described herein include one or more multimerization regions (e.g., heteropolymerization regions). In some embodiments, the heteropolymerization regions include dimerization, trimerization, or tetramerization regions.

[0505] In some embodiments, the multimerization regions are those described in International Publication No. 2017 / 081082, which are incorporated herein by reference in their entirety (e.g., SEQ ID NOs: 1116-1167, or fragments or variants thereof). Exemplary trimerization and tetramerization regions include, but are not limited to, the engineered leucine zipper, the fibrintin foldon domain derived from Enterobacterium phage T4, GCN4pll, GCN4-pll, and p53.

[0506] In some embodiments, the plasmodium polypeptide constructs provided herein can form trimer complexes. For example, the provided plasmodium polypeptide constructs may include a polymerizing region that enables the formation of a multimer complex, such as the trimer complex of the plasmodium polypeptide construct described herein. In some embodiments, the polymerizing region that enables the formation of a multimer complex includes a trimerizing region, for example, the trimerizing region described herein. In some embodiments, the plasmodium polypeptide construct includes a T4-fibrintin-derived "Foldon" trimerizing region, for example, to increase its immunogenicity. In some embodiments, the plasmodium polypeptide construct includes a polymerizing region comprising or consisting of the amino acid sequence GYIPEAPRDGQAYVRKDGEWVLLSTFL (SEQ ID NO: 78).

[0507] E. Linker In some embodiments, the plasmodium polypeptide constructs described herein include one or more linkers. In some embodiments, the linker is or includes 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids. In some embodiments, the linker is or includes about 30, 25, 20, 15, 10 or fewer amino acids. The linker may include any amino acid sequence and is not limited to any specific amino acids. In some embodiments, the linker includes one or more glycine (G) amino acids. In some embodiments, the linker includes one or more serine (S) amino acids. In some embodiments, the linker includes a glycine-serine linker. As used herein, “glycine-serine linker” refers to a linker that primarily (e.g., 80% or more) contains glycine and serine amino acids. In some embodiments, the linker includes amino acids selected based on cleavage predictors to produce a highly cleavable linker.

[0508] In some embodiments, the linker is or includes S-G4-S-G4-S. In some embodiments, the linker is or includes GSPGSGSGS (Sequence ID 79). In some embodiments, the linker is or includes GGSGGGGSGG (Sequence ID 80). In some embodiments, the linker is one shown in Table 6. In some embodiments, the linker is or includes the sequence described in whole in International Publication No. 2017 / 081082, which is incorporated herein by reference (see Sequence IDs 1509-1565, or fragments or variants thereof).

[0509] In some embodiments, the plasmodium polypeptide constructs described herein include a linker between the C-terminal region or a portion thereof and the transmembrane region. In some embodiments, the plasmodium polypeptide constructs described herein include a linker after a minor repeat sequence.

[0510] An exemplary linker is provided in Table 6 below:

[0511] [Table 13]

[0512] F. Self-assembly area In some embodiments, the plasmodium polypeptide constructs described herein include one or more self-assembling regions (e.g., self-assembling nanoparticle regions, e.g., heterogeneous self-assembling nanoparticle regions). In some embodiments, the self-assembling nanoparticle regions are ferritin regions. In some embodiments, the ferritin regions are derived from H. pylori. In some embodiments, the ferritin regions are, The sequence contains or consists of DIIKLLNEQVNKEMQSSNLYMSMSSWCYTHSLDGAGLFLFDHAAEEYEHAKKLIIFLNENNVPVQLTSISAPEHKFEGLTQIFQKAYEHEQHISESINNIVDHAIKSKDHATFNFLQWYVAEQHEEEVLFKDILDKIELIGNENHGLYLADQYVKGIAKSRKS (sequence number 88).

[0513] G. Tag Those skilled in the art will understand, upon reading this disclosure, that in some embodiments, one or more tags may be used when designing and testing constructs (e.g., plasmodium polypeptide constructs as described herein) in specific circumstances (e.g., in vitro, ex vivo, etc.). In some embodiments, one or more tags may be directly attached to the plasmodium polypeptide construct through peptide bonds (e.g., at the 5' or 3' end of the construct). In some embodiments, one or more tags may be attached to the plasmodium polypeptide construct through one or more linkers (e.g., one or more linkers as described herein). In some embodiments, one or more tags may be embedded internally within the plasmodium polypeptide construct, and one or more tags may be directly attached to the construct through peptide bonds (e.g., at the 5' and 3' ends of one or more tags). In some embodiments, one or more tags may be embedded internally within the plasmodium polypeptide construct, and one or more tags may be attached to the construct through one or more linkers (e.g., one or more linkers as described herein).

[0514] In some embodiments, the plasmodium polypeptide constructs described herein include one or more tags. In some embodiments, the plasmodium polypeptide constructs described herein include one or more detection tags (e.g., HiBit tags, HA tags, etc.). In some embodiments, the plasmodium polypeptide constructs described herein include or consist of a HiBit tag. In some embodiments, the HiBit tag has the amino acid sequence according to SEQ ID NO: 139.

[0515] Embodiments of H. plasmodium polypeptide constructs In some embodiments, the plasmodium polypeptide constructs described herein include one or more plasmodium Rh5 intrusion complex polypeptide regions or portions thereof. Exemplary combinations of regions are described below.

[0516] In some embodiments, the plasmodium polypeptide constructs described herein comprise one or more regions or portions of a Rh5 invasion complex polypeptide derived from Plasmodium falciparum, preferably Plasmodium falciparum isolate 3D7. In some embodiments, the Rh5 invasion complex polypeptide is selected from Plasmodium Rh5, Plasmodium CyRPA, Plasmodium Ripr, Plasmodium P113, Plasmodium TRAMP, and Plasmodium CSS.

[0517] 1. Rh5 structures In some embodiments, the plasmodium polypeptide constructs described herein include an ordered Rh5 domain derived from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.

[0518] In some embodiments, the Rh5 ordered domain includes the amino acid sequence at 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 with the amino acid sequence at positions 140-247 of SEQ ID NO: 1. In some embodiments, the Rh5 ordered domain includes the amino acid sequence at positions 140-247 of SEQ ID NO: 1, or an amino acid sequence at positions 140-247 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0519] In some embodiments, the Rh5 ordered domain includes the amino acid sequence at positions 146-247 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity with the amino acid sequence at positions 146-247 of SEQ ID NO: 1. In some embodiments, the Rh5 ordered domain includes the amino acid sequence at positions 146-247 of SEQ ID NO: 1, or an amino acid sequence at positions 146-247 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0520] In some embodiments, the Rh5 ordered domain includes the amino acid sequence at 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 with the amino acid sequence at positions 297-526 of SEQ ID NO: 1. In some embodiments, the Rh5 ordered domain includes the amino acid sequence at positions 297-526 of SEQ ID NO: 1, or an amino acid sequence at positions 297-526 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.

[0521] In some embodiments, the ordered domain of Rh5 can have the following structure: The sequence corresponding to amino acids 140-247 of sequence number 1 (ordered domain 140-247); The sequence corresponding to amino a...

Claims

1. A polyribonucleotide encoding a polypeptide, wherein the polypeptide comprises one or more plasmodium Rh5-invading complex polypeptides or their antigenic moieties.

2. The one or more plasmodium Rh5 entry complex polypeptides or their antigenic moieties (i) One or more plasmodium reticulocyte-binding protein homolog 5 (Rh5) polypeptides or their antigenic moieties, (ii) One or more plasmodium cysteine-rich protective antigen (CyRPA) polypeptides or their antigenic moieties, (iii) One or more plasmodium Rh5 interacting protein (Ripr) polypeptides or their antigenic moieties, (iv) One or more plasmodium P113 polypeptides or their antigenic moieties, (v) One or more plasmodium thrombospondin-associated apical merozoite protein (TRAMP) polypeptides or their antigenic moieties, (vi) One or more plasmodium cysteine-rich small secretory protein (CSS) polypeptides or their antigenic moieties The polyribonucleotide according to claim 1, comprising:

3. The one or more plasmodium Rh5 antigenic moieties described above (i) Amino acid sequence according to SEQ ID NOs: 118, 119, 124, or 126, (ii) The amino acid sequence according to SEQ ID NO: 120 or 128, or (iii) those combinations The polyribonucleotide according to claim 2, comprising:

4. The one or more plasmodium Rh5 antigenic moieties described above (i) Amino acid sequence according to SEQ ID NOs: 121, 122, 125, or 127, (ii) The amino acid sequence according to SEQ ID NO: 123 or 129, or (iii) those combinations The polyribonucleotide according to claim 2, comprising:

5. The one or more plasmodium Rh5 antigenic moieties described above (i) Amino acid sequence according to SEQ ID NOs: 124, 125, 126, or 127, (ii) The amino acid sequence according to SEQ ID NO: 128 or 129, or (iii) those combinations A polyribonucleotide according to claim 1 or 2, comprising:

6. The one or more plasmodium Rh5 antigenic moieties described above (i) an amino acid sequence having at least 85% sequence identity with the amino acid sequence of Sequence ID No. 130, (ii) an amino acid sequence having at least 85% sequence identity with the amino acid sequence of sequence numbers 118, 119, 121, 122, 124, 125, 126, or 127, (iii) An amino acid sequence having at least 85% sequence identity with the amino acid sequence of Sequence ID No. 131, (iv) an amino acid sequence having at least 85% sequence identity with the amino acid sequence of sequence number 120, 123, 128, or 129, or (v) combinations of those The polyribonucleotide according to claim 2, comprising:

7. The polyribonucleotide according to claim 1 or 2, wherein the polypeptide comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence from position 26 to 363 of SEQ ID NO: 94, 95, or 99.

8. The polyribonucleotide according to claim 1 or 2, wherein the polypeptide comprises an amino acid sequence having at least 85% sequence identity with the amino acid sequence from positions 26 to 526 of SEQ ID NO:

89.

9. The polyribonucleotide according to claim 1 or 2, wherein one or more plasmodium Rh5-invading complex polypeptides or their antigenic moieties comprise a plasmodium CyRPA polypeptide or its antigenic moiety.

10. The polyribonucleotide according to claim 1 or 2, wherein one or more plasmodium Rh5-invading complex polypeptides or their antigenic moieties comprise a plasmodium P113 polypeptide or its antigenic moiety.

11. The polyribonucleotide according to claim 1 or 2, wherein one or more plasmodium Rh5 intrusion complex polypeptides or their antigenic moieties comprise a plasmodium Ripr polypeptide or its antigenic moiety.

12. The polyribonucleotide according to claim 1 or 2, wherein one or more plasmodium Rh5-invading complex polypeptides or their antigenic moieties comprise a plasmodium TRAMP polypeptide or its antigenic moiety.

13. The polyribonucleotide according to claim 1 or 2, wherein one or more plasmodium Rh5-invading complex polypeptides or their antigenic moieties comprise a plasmodium CSS polypeptide or its antigenic moiety.

14. The polypeptide described above, (i) secretory signals, and (ii) Two plasmodium Rh5 ordered domains A polyribonucleotide according to claim 1 or 2, comprising:

15. The polypeptide described above, (i) secretory signals, (ii) Two plasmodium Rh5 ordered domains, (iii) Linker, and (iv) Multimerization domain A polyribonucleotide according to claim 1 or 2, comprising:

16. The polypeptide described above, (i) secretory signals, (ii) Two plasmodium Rh5 ordered domains, (iii) Linker, and (iv) Transmembrane region A polyribonucleotide according to claim 1 or 2, comprising:

17. The polypeptide described above, (i) secretory signals, (ii) Two plasmodium Rh5 ordered domains, (iii) Linker, and (iv) Self-assembling nanoparticle domains A polyribonucleotide according to claim 1 or 2, comprising:

18. The polyribonucleotide according to any one of claims 14 to 17, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO:

94.

19. The polyribonucleotide according to any one of claims 14 to 17, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO:

90.

20. The polyribonucleotide according to any one of claims 14 to 17, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO:

92.

21. The polyribonucleotide according to any one of claims 14 to 17, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of Sequence ID No.

96.

22. The polyribonucleotide according to any one of claims 14 to 17, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of Sequence ID No.

99.

23. The polyribonucleotide according to any one of claims 14 to 17, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of Sequence ID No.

98.

24. The polyribonucleotide according to any one of claims 14 to 17, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO:

101.

25. The polyribonucleotide according to any one of claims 14 to 17, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO:

100.

26. The polyribonucleotide according to any one of claims 14 to 17, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of Sequence ID No.

95.

27. The polyribonucleotide according to any one of claims 14 to 17, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of SEQ ID NO:

91.

28. The polyribonucleotide according to any one of claims 14 to 17, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of Sequence ID No.

93.

29. The polyribonucleotide according to any one of claims 14 to 17, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of Sequence ID No.

97.

30. The polypeptide described above, (i) secretory signals, (ii) Plasmodium Rh5 N-terminal disordered domain, (iii) Plasmodium Rh5 ordered domain, (iv) Plasmodium Rh5 linked disordered domain, and (v) C-terminal ordered domain of plasmodium Rh5 A polyribonucleotide according to claim 1 or 2, comprising:

31. The polyribonucleotide according to claim 30, wherein the polypeptide comprises or consists of an amino acid sequence having at least 85% sequence identity with the amino acid sequence of Sequence ID No.

89.

32. An RNA construct, in the order from 5' to 3', (i) 5' UTR, (ii) The polyribonucleotide according to any one of claims 1 to 532, (iii) 3'UTR, and (iv) PolyA tail arrangement RNA constructs that include this.

33. A composition comprising one or more polyribonucleotides as described in any one of claims 1 to 31.

34. A composition comprising one or more RNA constructs as described in claim 32.

35. Further comprising lipid nanoparticles, polyplexes (PLX), lipid-modified polyplexes (LPLX), or liposomes, The one or more polyribonucleotides are completely or partially encapsulated within the lipid nanoparticles, polyplexes (PLX), lipid-modified polyplexes (LPLX), or liposomes. The composition according to claim 33 or 34.

36. A pharmaceutical composition comprising the composition according to claim 35 and at least one pharmaceutically acceptable excipient.

37. It is a combination, (i) A first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more plasmodium Rh5 polypeptides or their antigenic moieties, and (ii) A second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, and the second polypeptide is One or more CyRPA polypeptides or their antigenic moieties, One or more Ripr polypeptides or their antigenic moieties, One or more P113 polypeptides or their antigenic moieties, One or more TRAMP polypeptides or their antigenic moieties, One or more CSS polypeptides or their antigenic moieties, those combinations A second pharmaceutical composition comprising one or more plasmodium Rh5-invading complex polypeptides selected from A combination that includes this.

38. It is a combination, (i) A first pharmaceutical composition comprising a first polyribonucleotide according to any one of claims 1 to 31, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more plasmodium Rh5-invading complex polypeptides or their antigenic moieties, and (ii) A second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, and the second polypeptide comprises one or more plasmodium T cell antigens. A combination that includes this.

39. It is a combination, (i) A first pharmaceutical composition comprising a first polyribonucleotide according to any one of claims 1 to 31, wherein the first polyribonucleotide encodes a first polypeptide, and the first polypeptide comprises one or more plasmodium Rh5-invading complex polypeptides or their antigenic moieties, and (ii) A second pharmaceutical composition comprising a second polyribonucleotide, wherein the second polyribonucleotide encodes a second polypeptide, and the second polypeptide comprises one or more plasmodium CSP polypeptides or their antigenic moieties. A combination that includes this.

40. A method comprising administering the polyribonucleotide described in any one of claims 1 to 31.

41. A method comprising administering the RNA construct described in claim 32 to a target.

42. A method comprising administering the composition described in any one of claims 33 to 35 to a subject.

43. A method comprising administering one or more doses of the pharmaceutical composition described in claim 36 to a target.

44. The pharmaceutical composition according to claim 36, for use in the treatment of malaria infection, comprising administering one or more doses of the pharmaceutical composition to the target.

45. The pharmaceutical composition according to claim 36, for use in the prevention of malaria infection, comprising administering one or more doses of the pharmaceutical composition to a target.

46. A method comprising administering the combination described in any one of claims 37 to 39 to a target.

47. Use of the pharmaceutical composition according to claim 36 in the treatment of malaria infection.

48. Use of the pharmaceutical composition according to claim 36 in the prevention of malaria infection.

49. Use of the pharmaceutical composition according to claim 36 in inducing an antimalarial immune response in a subject.

50. A polypeptide encoded by a polyribonucleotide according to any one of claims 1 to 31.

51. A polypeptide encoded by the RNA construct according to claim 32.

52. A host cell containing polyribonucleotides according to any one of claims 1 to 31.

53. A host cell comprising the RNA construct described in claim 32.

54. A host cell comprising the polypeptide according to claim 50 or 51.

Images