EP4750488A1 - Compositions pour l'administration d'antigènes de plasmodium et méthodes associées - Google Patents

Compositions pour l'administration d'antigènes de plasmodium et méthodes associées

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Publication number
EP4750488A1
EP4750488A1 EP24752314.5A EP24752314A EP4750488A1 EP 4750488 A1 EP4750488 A1 EP 4750488A1 EP 24752314 A EP24752314 A EP 24752314A EP 4750488 A1 EP4750488 A1 EP 4750488A1
Authority
EP
European Patent Office
Prior art keywords
plasmodium
amino acid
polypeptide
acid sequence
csp
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP24752314.5A
Other languages
German (de)
English (en)
Inventor
Ugur Sahin
Asaf PORAN
Daniel Abram Rothenberg
Stephanie ERBAR
Annette VOGEL
Patricia DOS SANTOS MEIRELES
John SROUJI
Anja DOKIC
Thorsten Klamp
Charles JENNISON
Theresa ADDONA
Raquel Mariska FURTADO
Thomas ZIEGENHALS
Charles Lefco DULBERGER
Alexandra WALLS
Gavin PALOWITCH
Lynda Maria STUART
Karla-Gerlinde SCHRAUT
Svenja GROBE
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BioNTech SE
Original Assignee
BioNTech SE
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Filing date
Publication date
Application filed by BioNTech SE filed Critical BioNTech SE
Publication of EP4750488A1 publication Critical patent/EP4750488A1/fr
Pending legal-status Critical Current

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/0003—Invertebrate antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/002—Protozoa antigens
    • A61K39/015—Hemosporidia antigens, e.g. Plasmodium antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
    • A61P33/06—Antimalarials
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/44—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from protozoa
    • C07K14/445—Plasmodium
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/53—DNA (RNA) vaccination
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55555—Liposomes; Vesicles, e.g. nanoparticles; Spheres, e.g. nanospheres; Polymers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
    • A61K2039/572—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 cytotoxic response
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
    • A61K2039/575—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 humoral response
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/64—Medicinal preparations containing antigens or antibodies characterised by the architecture of the carrier-antigen complex, e.g. repetition of carrier-antigen units
    • A61K2039/645—Dendrimers; Multiple antigen peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/70—Multivalent vaccine
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present disclosure provides technologies (e.g., compositions, methods, etc.) for delivery of Plasmodium antigens.
  • the present disclosure provides pharmaceutical compositions (e.g., immunogenic compositions, e.g., vaccines) for delivering particular Plasmodium polypeptide constructs to a subject (e.g., a patient) and related technologies (e.g., methods).
  • compositions e.g., immunogenic compositions, e.g., vaccines
  • Plasmodium polypeptide constructs can comprise one or more Plasmodium polypeptide or antigenic portions thereof.
  • the present disclosure provides Plasmodium vaccine compositions and related technologies (e.g., methods).
  • the present disclosure provides particular pharmaceutical composition (e.g., immunogenic composition, e.g., vaccine) formats including, for example, polyribonucleotides comprising particular elements and/or sequences useful for delivery of Plasmodium antigens.
  • the present disclosure provides a variety of insights and technologies related to such Plasmodium antigen constructs and vaccine (e.g., RNA vaccine) compositions.
  • the present disclosure provides technologies for preventing, characterizing, treating, and/or monitoring malaria outbreaks and/or infections including, as noted, various nucleic acid constructs and encoded proteins, as well as agents (e.g., antibodies) that bind to such proteins, and compositions that comprise and/or deliver them.
  • technologies e.g., compositions and methods for augmenting, inducing, promoting, enhancing and/or improving an immune response against a Plasmodium parasite.
  • technologies described herein are designed to act as immunological boost to a primary vaccine, such as a vaccine directed to antigen(s) and/or epitope(s) of a Plasmodium parasite.
  • the present disclosure further recognizes that an approach comprising the delivery of one or more Plasmodium polypeptides or antigenic portions thereof that are present and/or exposed at various time points in a Plasmodium life cycle may elicit a more robust immune response.
  • Plasmodium polypeptides or antigenic portions thereof that will generate an immune response that targets Plasmodium parasites at an early life cycle stage, e.g., (1) in the asymptomatic infection stage from the deposition of the parasites in the skin until the infection of hepatocytes (see FIG.1, 1 – Sporozoite), and/or (2) after infection of a subject’s hepatocytes (see FIG.1, 2 – Liver Stage), and/or (3) after egress from hepatocytes but before infecting a subject’s erythrocytes (see FIG.1, 3 – Pre-invasion).
  • an early life cycle stage e.g., (1) in the asymptomatic infection stage from the deposition of the parasites in the skin until the infection of hepatocytes (see FIG.1, 1 – Sporozoite), and/or (2) after infection of a subject’s hepatocytes (see FIG.1, 2 – Liver Stage), and/or (3) after egress from
  • Preventing the symptomatic phase of a Plasmodium parasite e.g., killing the parasites prior to infecting erythrocytes, can help prevent disease and death of a subject, as well as mitigate onward transmission.
  • the approach may enable interruption of transmission and aid in malaria elimination.
  • the present disclosure provides, among other things, combinations comprising more than one Plasmodium polypeptide construct that each include one or more Plasmodium polypeptides or antigenic fragments thereof, wherein the more than one Plasmodium polypeptide constructs comprise Plasmodium polypeptides or antigenic fragments thereof that are expressed at different stages of the asymptomatic phase of infection, such as the initial sporozoite stage and the liver stage of infection.
  • Plasmodium polypeptide constructs that include Plasmodium polypeptides or antigenic fragments thereof that are expressed at different stages within the asymptomatic phase (such as sporozoite and liver stage) may prevent erythrocyte infection by parasites and thereby prevent symptomatic malaria disease and disrupting onwards transmission.
  • Plasmodium polypeptide constructs described herein provide a combination that acts as a pre-erythrocytic vaccine.
  • targeting two or more stages of the parasitic infection can provide additional layers of immune protection for a subject, e.g., fewer Plasmodium sporozoites invading the liver, leading to fewer infected hepatocytes that need to be eliminated.
  • a combination as described herein includes two or more Plasmodium polypeptide constructs, wherein the Plasmodium polypeptide constructs comprise Plasmodium polypeptides or antigenic fragments thereof that are expressed at different stages within the asymptomatic phase (such as sporozoite and liver stage).
  • the two or more Plasmodium polypeptide constructs do not interfere with each other (e.g., the elicitation of an immune response by one Plasmodium polypeptide construct does not interfere with the elicitation of an immune response by another of the Plasmodium polypeptide constructs.
  • the presence of two or more Plasmodium polypeptide constructs in a combination does not abolish an immunogenic effect of one or more antigens on a first Plasmodium polypeptide constructs that is otherwise present in the absence of a second Plasmodium polypeptide constructs.
  • the lack of interference by one Plasmodium polypeptide construct with another is observed regardless of whether there are three or more (e.g., three, four, five, six, etc.) present in a combination.
  • a combination of two or more Plasmodium polypeptide constructs elicits a humoral and/or cellular immune response.
  • each Plasmodium polypeptide construct elicits a humoral and/or cellular immune response.
  • the present disclosure provides the insight that antigens or epitopes that are present on the surface of cells, particularly the Plasmodium cell surface, may be able to induce a B cell response, while antigens or epitopes that are not surface exposed or are minimally exposed may still be able to induce a T cell response.
  • the resulting combined B cell and T cell response may be important for developing strong immune protection against current and future Plasmodium infections.
  • the present disclosure provides compositions for delivering antigens or epitopes that can be useful prophylactically or therapeutically.
  • the present disclosure also provides the insight that certain, multi-prong approaches can be useful for providing protection against Plasmodium infections (e.g., malaria).
  • a combination as described herein can comprise a first pharmaceutical composition comprising a first polyribonucleotide and a second pharmaceutical composition comprising a second polyribonucleotide.
  • a first polyribonucleotide encodes a first polypeptide that comprises one or more Plasmodium T-cell antigens.
  • a second polyribonucleotide encodes a second polypeptide that comprises one or more Plasmodium polypeptides or antigenic portions thereof.
  • a first polypeptide comprises an amino acid sequence with at least 85% identity to an amino acid sequence according to any one of SEQ ID NOs 167, 170, 173, 176, 179, 182, 185, 188, 191, 194, 197, 200, 203, 206, 209, 212, 215, 218, and 221; and a second polyribonucleotide comprises an amino acid sequence with at least 85% identity to an amino acid sequence according to any one of SEQ ID NOs: 5, 8, 10, 12, 15, 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, 60, 63, 66, 69, 72, 75, 78, 81, 84, 87, 90, 93, 96, 99, 102, 105 , 107-112, 117, 122, 125, 130, 135, 138, and 141.
  • a first polypeptide comprises (i) an antigenic Plasmodium CSP polypeptide fragment, (ii) an antigenic Plasmodium TRAP polypeptide fragment, (iii) an antigenic Plasmodium UIS3 polypeptide fragment, (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment, and (v) an antigenic Plasmodium LSAP2 polypeptide fragment.
  • a second polyribonucleotide comprises (i) a secretory signal, (ii) a Plasmodium CSP N-terminal end region, (iii) a Plasmodium CSP junction region, (iv) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (v) a Plasmodium CSP C-terminal region, a Plasmodium CSP C-terminal region variant, or an antigenic portion thereof, (vi) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region, a Plasmodium CSP C-terminal region variant, or an antigenic portion thereof, (vii) a linker, and (viii) a transmembrane region, and wherein the second polypeptide does not comprise any of (a) a Plasmodium CSP N-terminal region or portion thereof, and (b) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polyribonucleotide comprises (i) a secretory signal, (ii) a Plasmodium CSP N-terminal end region, (iii) a Plasmodium CSP junction region, (iv) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (v) a Plasmodium CSP C-terminal region, (vi) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region, (vii) a linker, and (viii) a transmembrane region, and wherein the second polypeptide does not comprise any of (a) a Plasmodium CSP N-terminal region or portion thereof, and (b) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a first polypeptide comprises (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment, (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment, (iii) an antigenic Plasmodium LISP-2 polypeptide fragment, and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a second polyribonucleotide comprises (i) a secretory signal, (ii) a Plasmodium CSP N-terminal region, (iii) a Plasmodium CSP N-terminal end region, (iv) a Plasmodium CSP junction region, (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (vi) a Plasmodium CSP major repeat region, (vii) a Plasmodium CSP C- terminal region, a Plasmodium CSP C-terminal region variant, or an antigenic portion thereof, and a transmembrane region.
  • a first polypeptide comprises (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment, (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment, (iii) an antigenic Plasmodium LISP-2 polypeptide fragment, and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a second polyribonucleotide comprises (i) a secretory signal, (ii) a Plasmodium CSP N-terminal region, (iii) a Plasmodium CSP N-terminal end region, (iv) a Plasmodium CSP junction region, (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (vi) a Plasmodium CSP major repeat region, (vii) a Plasmodium CSP C- terminal region, and a transmembrane region.
  • a combination includes a first pharmaceutical composition and a second pharmaceutical composition.
  • a first pharmaceutical composition comprises a first polyribonucleotide. In some embodiments, a first polyribonucleotide encodes a first polypeptide. In some embodiments, a first polypeptide comprises one or more Plasmodium T-cell antigens. In some embodiments, a second pharmaceutical composition comprises a second polyribonucleotide. In some embodiments, a second polyribonucleotide encodes a second polypeptide. In some embodiments, a second polypeptide comprises one or more Plasmodium polypeptide or antigenic portions thereof.
  • a combination includes: (i) a first pharmaceutical composition comprising a first polyribonucleotide, wherein the first polyribonucleotide encodes a first polypeptide, and a first polypeptide comprises one or more Plasmodium T-cell antigens 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 polypeptide or antigenic portions thereof.
  • a first polypeptide comprises at least 10 amino acids and at most 1100 amino acids.
  • a first polypeptide comprises at least 10 amino acids and at most 500 amino acids.
  • one or more Plasmodium T-cell antigens comprised in a first polypeptide comprise at least 2 Plasmodium T-cell antigens.
  • one or more Plasmodium T-cell antigens comprised in a first polypeptide comprise at most 10 Plasmodium T-cell antigens.
  • one or more Plasmodium T-cell antigens comprised in a first polypeptide comprise at least 2 and at most 10 Plasmodium T-cell antigens.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium CSP polypeptide fragment. In some embodiments, one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LSA-1 polypeptide fragment. In some embodiments, one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium TRAP polypeptide fragment. In some embodiments, one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LSAP2 polypeptide fragment.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium UIS3 polypeptide fragment. In some embodiments, one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium ETRAMP10.3 polypeptide fragment. In some embodiments, one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LISP-1 polypeptide fragment. In some embodiments, one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LISP-2 polypeptide fragment.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LSA-3 polypeptide fragment. In some embodiments, one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LSA-1(a) polypeptide fragment. In some embodiments, one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LSA-1(b) polypeptide fragment.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise two or more of: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium LSA-1 polypeptide fragment; (iii) an antigenic Plasmodium TRAP polypeptide fragment; (iv) an antigenic Plasmodium LSAP2 polypeptide fragment; (v) an antigenic Plasmodium UIS3 polypeptide fragment; (vi) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (vii) an antigenic Plasmodium LISP-1 polypeptide fragment; (viii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (ix) an antigenic Plasmodium LSA-3 polypeptide fragment.
  • a first polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 179.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA-3 polypeptide fragment; (vii) an antigenic Plasmodium LSA-1(a) polypeptide fragment; and (viii) an antigenic Plasmodium LSA-1(b) polypeptide fragment.
  • a first polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 182.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA- 1(a) polypeptide fragment; (vii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (viii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (ix) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a first polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 188.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA- 1(a) polypeptide fragment; (vii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; and (viii) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a first polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 191.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LISP- 2 polypeptide fragment; and (vii) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a first polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 194.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA- 1(b) polypeptide fragment; and (vii) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a first polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 197.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA- 1(a) polypeptide fragment; (vii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (viii) an antigenic Plasmodium LISP-2 polypeptide fragment; (ix) an antigenic Plasmodium LISP-1 polypeptide fragment; and (x) an antigenic Plasmodium LSA-3 polypeptide fragment.
  • a first polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 200.
  • one or more Plasmodium T cell antigens comprised in the first polypeptide comprise: (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (iii) an antigenic Plasmodium LISP-2 polypeptide fragment; (iv) an antigenic Plasmodium LISP- 1 polypeptide fragment; and (v) an antigenic Plasmodium LSA-3 polypeptide fragment.
  • a first polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 212.
  • one or more Plasmodium T cell antigens comprised in the first polypeptide comprise: (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (iii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a first polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 209.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium CSP polypeptide fragment, and wherein the antigenic Plasmodium CSP polypeptide fragment comprises a Plasmodium CSP N-terminal region.
  • an antigenic Plasmodium CSP polypeptide fragment further comprises a Plasmodium CSP N-terminal end region. In some embodiments, an antigenic Plasmodium CSP polypeptide fragment further comprises a Plasmodium CSP junction region. In some embodiments, an antigenic Plasmodium CSP polypeptide fragment comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 437.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide do not comprise an antigenic Plasmodium berghei CSP polypeptide fragment.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LSA-1(a) polypeptide fragment, and wherein the antigenic Plasmodium LSA-1(a) polypeptide fragment comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 447.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LSA-1(b) polypeptide fragment, and wherein the antigenic Plasmodium LSA-1(b) polypeptide fragment comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 457.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium TRAP polypeptide fragment, and wherein the antigenic Plasmodium TRAP polypeptide fragment comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 472.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LSAP2 polypeptide fragment, and wherein the antigenic Plasmodium LSAP2 polypeptide fragment comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 497.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium UIS3 polypeptide fragment, and wherein the antigenic Plasmodium UIS3 polypeptide fragment comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 510.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium ETRAMP10.3 polypeptide fragment, and wherein the antigenic Plasmodium ETRAMP10.3 polypeptide fragment comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 516.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LISP-1 polypeptide fragment, and wherein the antigenic Plasmodium LISP-1 polypeptide fragment comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 252.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LISP-2 polypeptide fragment, and wherein the antigenic Plasmodium LISP-2 polypeptide fragment comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 533.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide comprise an antigenic Plasmodium LSA-3 polypeptide fragment, and wherein the antigenic Plasmodium LSA-3 polypeptide fragment comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 543.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide each comprise one or more T cell epitopes.
  • a first polypeptide further comprises an MHC class I trafficking signal (MITD).
  • MITD MHC class I trafficking signal
  • a MITD is located at the C-terminal end comprised in the first polypeptide.
  • a MITD comprises or consists of an amino acid sequence according to SEQ ID NO: 561.
  • a first polypeptide comprises a secretory signal.
  • a secretory signal comprises or consists of a Plasmodium secretory signal.
  • a Plasmodium secretory signal comprises or consists of a Plasmodium CSP secretory signal.
  • a Plasmodium CSP secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 332.
  • a secretory signal comprises or consists of a heterologous secretory signal.
  • a heterologous secretory signal comprises or consists of a non-human secretory signal.
  • a heterologous secretory signal comprises or consists of a viral secretory signal.
  • a viral secretory signal comprises or consists of an HSV secretory signal.
  • a HSV secretory signal comprises or consists of an HSV-1 or HSV-2 secretory signal.
  • a HSV secretory signal comprises or consists of an HSV glycoprotein D (gD) secretory signal.
  • a HSV gD secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 314.
  • a HSV gD secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 320.
  • a viral secretory signal comprises or consists of an Ebola virus secretory signal.
  • an Ebola virus secretory signal comprises or consists of an Ebola virus spike glycoprotein (SGP) secretory signal.
  • SGP Ebola virus spike glycoprotein
  • an Ebola virus SGP secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 335.
  • a secretory signal is located at the N-terminus of the polypeptide.
  • a first polypeptide does not comprise a secretory signal.
  • a polypeptide comprises one or more linkers.
  • one or more linkers comprise one or more glycine-serine linkers.
  • one or more linkers comprise at least one linker comprising an amino acid sequence according to SEQ ID NO: 404.
  • one or more linkers comprise at least one linker comprising an amino acid sequence according to SEQ ID NO: 411.
  • one or more linkers comprise at least one linker comprising an amino acid sequence according to SEQ ID NO: 408. In some embodiments, one or more linkers comprise at least one linker comprising an amino acid sequence according to SEQ ID NO: 412. In some embodiments, a polypeptide comprises a linker between two Plasmodium T-cell antigens. [0042] In some embodiments, a polypeptide comprises a transmembrane region. In some embodiments, a transmembrane region comprises or consists of a Plasmodium transmembrane region.
  • a Plasmodium transmembrane region comprises or consists of a Plasmodium CSP glycosylphosphatidylinositol (GPI) anchor region.
  • a Plasmodium CSP GPI anchor region comprises or consists of an amino acid sequence according to SEQ ID NO: 385.
  • a transmembrane region comprises or consists of a heterologous transmembrane region.
  • a heterologous transmembrane region does not comprise a hemagglutinin transmembrane region.
  • a heterologous transmembrane region comprises or consists of a non-human transmembrane region.
  • a heterologous transmembrane region comprises or consists of a viral transmembrane region. In some embodiments, a heterologous transmembrane region comprises or consists of an HSV transmembrane region. In some embodiments, a HSV transmembrane region comprises or consists of an HSV-1 or HSV-2 transmembrane region. In some embodiments, a HSV transmembrane region comprises or consists of an HSV gD transmembrane region. In some embodiments, a HSV gD transmembrane region comprises or consists of an amino acid sequence according to SEQ ID NO: 379.
  • a transmembrane region comprises or consists of a human transmembrane region.
  • a human transmembrane region comprises or consists of a human decay accelerating factor glycosylphosphatidylinositol (hDAF-GPI) anchor region.
  • hDAF-GPI anchor region comprises or consists of an amino acid sequence according to SEQ ID NO: 382.
  • a polypeptide does not comprise a transmembrane region. [0043] In some embodiments, a polypeptide does not comprise an antigenic fragment of a bacterial polypeptide.
  • a polypeptide does not comprise an antigenic bacillus Calmette-Guérin (BCG) polypeptide fragment, optionally wherein the antigenic BCG polypeptide fragment comprises an amino acid sequence according to SEQ ID NO: 416.
  • a polypeptide does not comprise an antigenic tetanus toxin (TT) polypeptide fragment, optionally wherein the antigenic TT polypeptide fragment comprises an amino acid sequence according to SEQ ID NO: 417.
  • one or more Plasmodium T cell antigens comprised in a first polypeptide do not comprise an antigenic Plasmodium sporozoite threonine–asparagine-rich protein (STARP) polypeptide fragment, and optionally wherein the antigenic Plasmodium STARP polypeptide fragment comprises an amino acid sequence according to SEQ ID NO: 418.
  • one or more Plasmodium polypeptide or antigenic portions thereof comprised in a second polypeptide are one or more Plasmodium CSP polypeptide regions or antigenic portions thereof.
  • each of the one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise 10 or more contiguous amino acids of the amino acid sequence according to SEQ ID NO: 1.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise one or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise one or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), and wherein the second polypeptide does not comprise the amino acid sequence of NPNA (SEQ ID NO: 228). In some embodiments, one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise two or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise five or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223). In some embodiments, one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise between two and twelve repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223). In some embodiments, one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise exactly three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise between four and twelve repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise exactly eight repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise exactly nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • repeats of the amino acid sequence of NANPNVDP are all contiguous with each other. In some embodiments, repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223) are not all contiguous with each other.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise three portions of a Plasmodium CSP polypeptide, wherein each portion comprises three contiguous repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), and wherein each of the portions are not contiguous with each other.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise four portions of a Plasmodium CSP polypeptide, and wherein each portion comprises two contiguous repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • one or more Plasmodium CSP polypeptide regions or antigenic portions thereof comprise at least two repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • one or more Plasmodium CSP polypeptide regions or antigenic portions thereof comprise two to eighteen repeats of the amino acid sequence of NANP.
  • one or more Plasmodium CSP polypeptide regions or antigenic portions thereof comprise a Plasmodium CSP C-terminal region, a Plasmodium CSP C-terminal region variant, or an antigenic portion thereof.
  • one or more Plasmodium CSP polypeptide regions or antigenic portions thereof comprise: (i) at least two repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (ii) two to eighteen repeats of the amino acid sequence of NANP; and (iii) a Plasmodium CSP C-terminal region, a Plasmodium CSP C-terminal region variant, or an antigenic portion thereof.
  • one or more Plasmodium CSP polypeptide regions or antigenic portions thereof comprise, in N-terminal to C-terminal order: (i) at least two repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (ii) two to eighteen repeats of the amino acid sequence of NANP; and (iii) a Plasmodium CSP C-terminal region, a Plasmodium CSP C-terminal region variant, or an antigenic portion thereof.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise one or more Plasmodium CSP C-terminal regions or antigenic portions thereof.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise one or more Plasmodium CSP C-terminal region variants, or antigenic portions thereof.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise exactly one Plasmodium CSP C-terminal region.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise exactly one Plasmodium CSP C-terminal region, and wherein the Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 235.
  • one or more Plasmodium CSP polypeptide regions or antigenic portions thereof comprise an antigenic portion of a Plasmodium CSP C-terminal region.
  • one or more Plasmodium CSP polypeptide regions or antigenic portions thereof comprise a Plasmodium CSP C-terminal region variant. In some embodiments, one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise two or more antigenic portions of a Plasmodium CSP C-terminal region.
  • an antigenic portion of a Plasmodium CSP C-terminal region comprises an amino acid sequence according SEQ ID NO: 261, wherein X3 is N or K, X4 is K, I, or R, and X5 is N or Y.
  • an antigenic portion of a Plasmodium CSP C-terminal region comprises an amino acid sequence according SEQ ID NO: 262, wherein X3 is N or K, X4 is K, I, or R, and X5 is N or Y. In some embodiments, an antigenic portion of a Plasmodium CSP C-terminal region comprises an amino acid sequence according SEQ ID NO: 263, wherein X3 is N or K, X4 is K, I, or R, and X5 is N or Y.
  • an antigenic portion of a Plasmodium CSP C-terminal region comprises an amino acid sequence according SEQ ID NO: 264, wherein X1X2 is EK or KE, X3 is N or K, X4 is K, I, or R, and X5 is N or Y. In some embodiments, an antigenic portion of a Plasmodium CSP C-terminal region comprises an amino acid sequence according SEQ ID NO: 265, wherein X1X2 is EK or KE, X3 is N or K, X4 is K, I, or R, and X5 is N or Y.
  • an antigenic portion of a Plasmodium CSP C-terminal region comprises an amino acid sequence according SEQ ID NO: 266, wherein X1X2 is EK or KE, X3 is N or K, X4 is K, I, or R, and X5 is N or Y. In some embodiments, an antigenic portion of a Plasmodium CSP C-terminal region comprises an amino acid sequence according SEQ ID NO: 267. In some embodiments, an antigenic portion of a Plasmodium CSP C-terminal region or a Plasmodium CSP C-terminal region variant comprises an amino acid sequence according SEQ ID NO: 992.
  • an antigenic portion of a Plasmodium CSP C-terminal region or a Plasmodium CSP C-terminal region variant comprises an amino acid sequence according SEQ ID NO: 993.
  • one or more Plasmodium CSP polypeptide regions or antigenic portions thereof comprise a Plasmodium CSP C-terminal region variant or antigenic portion thereof.
  • a Plasmodium CSP C-terminal region variant or antigenic portion thereof comprises one or more amino acid substitutions, insertions, or deletions.
  • a Plasmodium CSP C-terminal region variant or antigenic portion thereof comprises one or more amino acid substitutions.
  • a Plasmodium CSP C-terminal region variant or antigenic portion thereof comprises one or more amino acid substitutions, wherein the one or more amino acid substitutions comprise S301N, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a Plasmodium CSP C-terminal region variant or antigenic portion thereof comprises one or more amino acid substitutions, wherein the one or more amino acid substitutions comprise K317E, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • a Plasmodium CSP C-terminal region variant or antigenic portion thereof comprises one or more amino acid substitutions, wherein the one or more amino acid substitutions comprise E318Q, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • a Plasmodium CSP C- terminal region variant or antigenic portion thereof comprises one or more amino acid substitutions, wherein the one or more amino acid substitutions comprise N321K, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • a Plasmodium CSP C-terminal region variant or antigenic portion thereof comprises one or more amino acid substitutions, wherein the one or more amino acid substitutions comprise E357Q, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, a Plasmodium CSP C-terminal region variant or antigenic portion thereof comprises one or more amino acid substitutions, wherein the one or more amino acid substitutions comprise A361E, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • a Plasmodium CSP C-terminal region variant or antigenic portion thereof comprises one or more amino acid substitutions, wherein the one or more amino acid substitutions comprise S301N K317E, E318Q, N321K, E357Q, A361E, or any combination thereof, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • a Plasmodium CSP C-terminal region variant or antigenic portion thereof comprises one or more amino acid substitutions, wherein the one or more amino acid substitutions comprise S301N, K317E, E318Q, N321K, E357Q, and A361E, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • a Plasmodium CSP C-terminal region variant comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 994.
  • a second polypeptide comprises one or more portions of the Plasmodium CSP C- terminal region, wherein each of the one or more portions comprise or consist of an amino acid sequence according to SEQ ID NO: 244.
  • a second polypeptide comprises one or more portions of the Plasmodium CSP C-terminal region, wherein each of the one or more portions comprise or consist of an amino acid sequence according to SEQ ID NO: 248. In some embodiments, a second polypeptide comprises one or more portions of the Plasmodium CSP C-terminal region, wherein each of the one or more portions comprise or consist of an amino acid sequence according to SEQ ID NO: 262. In some embodiments, a second polypeptide comprises one or more portions of the Plasmodium CSP C-terminal region, wherein each of the one or more portions comprise or consist of an amino acid sequence according to SEQ ID NO: 256.
  • a second polypeptide comprises one or more portions of the Plasmodium CSP C-terminal region, wherein each of the one or more portions comprise or consist of: (i) an amino acid sequence according to SEQ ID NO: 244; (ii) an amino acid sequence according to SEQ ID NO: 248; (iii) an amino acid sequence according to SEQ ID NO: 262; (iv) an amino acid sequence according to SEQ ID NO: 256; or (v) a combination thereof. [0058] In some embodiments, a second polypeptide comprises one portion of the Plasmodium CSP C-terminal region, wherein the portion comprises or consists of an amino acid sequence according to SEQ ID NO: 244.
  • a second polypeptide comprises one portion of the Plasmodium CSP C-terminal region, wherein the portion comprises or consists of an amino acid sequence according to SEQ ID NO: 248. In some embodiments, a second polypeptide comprises one portion of the Plasmodium CSP C-terminal region, wherein the portion comprises or consists of an amino acid sequence according to SEQ ID NO: 262. In some embodiments, a second polypeptide comprises one portion of the Plasmodium CSP C-terminal region, wherein the portion comprises or consists of an amino acid sequence according to SEQ ID NO: 256.
  • a second polypeptide comprises one portion of the Plasmodium CSP C-terminal region, wherein the portion comprises or consists of: (i) an amino acid sequence according to SEQ ID NO: 244; (ii) an amino acid sequence according to SEQ ID NO: 248; (iii) an amino acid sequence according to SEQ ID NO: 262; (iv) an amino acid sequence according to SEQ ID NO: 256; or (v) a combination thereof.
  • a second polypeptide comprises one or more portions of the Plasmodium CSP C-terminal region, wherein the one or more portions collectively comprise or consist of: (i) an amino acid sequence according to SEQ ID NO: 244; (ii) an amino acid sequence according to SEQ ID NO: 248; (iii) an amino acid sequence according to SEQ ID NO: 262; and (iv) an amino acid sequence according to SEQ ID NO: 256.
  • an antigenic portion of a Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 259.
  • a second polypeptide comprises a serine immediately following a Plasmodium CSP C-terminal region.
  • a second polypeptide comprises a serine-valine sequence immediately following a Plasmodium CSP C-terminal region.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise one or more Plasmodium CSP junction regions, portions thereof, or variants thereof. In some embodiments, one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise two or more Plasmodium CSP junction regions or portions thereof. In some embodiments, one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise exactly one Plasmodium CSP junction region. In some embodiments, one Plasmodium CSP junction region consists of an amino acid sequence according to SEQ ID NO: 272.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise one or more portions of a Plasmodium CSP junction region.
  • one or more portions of a Plasmodium CSP junction region comprise a deletion of K93, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • one or more portions of a Plasmodium CSP junction region comprise a deletion of L94, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • one or more portions of a Plasmodium CSP junction region comprise a deletion of K95, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • one or more portions of a Plasmodium CSP junction region comprise a deletion of Q96, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, one or more portions of a Plasmodium CSP junction region comprise a deletion of P97, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, one or more portions of a Plasmodium CSP junction region comprise a deletion of K93, L94, K95, Q96, P97, or a combination thereof, and wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • one or more portions of a Plasmodium CSP junction region comprise a deletion of K93, L94, K95, and Q96, and wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, one or more portions of a Plasmodium CSP junction region comprise a deletion of K93, L94, K95, Q96 and P97, and wherein the amino acid numbering is relative to SEQ ID NO: 1. [0063] In some embodiments, each portion of a Plasmodium CSP junction region comprises or consists of an amino acid sequence according to SEQ ID NO: 275. In some embodiments, each portion of a Plasmodium CSP junction region comprises or consists of an amino acid sequence according to SEQ ID NO: 277.
  • two or more Plasmodium CSP junction regions consist of an amino acid sequence according to SEQ ID NO: 272.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise two or more portions of a Plasmodium CSP junction region.
  • two or more portions of a Plasmodium CSP junction region comprise a deletion of K93, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • two or more portions of a Plasmodium CSP junction region comprise a deletion of L94, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • two or more portions of a Plasmodium CSP junction region comprise a deletion of K95, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, two or more portions of a Plasmodium CSP junction region comprise a deletion of Q96, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, two or more portions of a Plasmodium CSP junction region comprise a deletion of P97, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • two or more portions of a Plasmodium CSP junction region comprise a deletion of K93, L94, K95, Q96, P97, or a combination thereof, and wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • two or more portions of a Plasmodium CSP junction region comprise a deletion of K93, L94, K95, and Q96, and wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • two or more portions of a Plasmodium CSP junction region comprise a deletion of K93, L94, K95, Q96 and P97, and wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • each portion of a Plasmodium CSP junction region comprises or consists of an amino acid sequence according to SEQ ID NO: 275. In some embodiments, each portion of a Plasmodium CSP junction region comprises or consists of an amino acid sequence according to SEQ ID NO: 277. [0066] In some embodiments, one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise one or more Plasmodium CSP junction region variants. In some embodiments, a Plasmodium CSP junction region variant comprises one or more substitution mutations. In some embodiments, one or more substitution mutations comprise a K93A mutation, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • one or more substitution mutations comprise a L94A mutation, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, one or more substitution mutations comprise a K93A mutation, an L94A mutation, or both, wherein the amino acid numbering is relative to SEQ ID NO: 1. In some embodiments, each Plasmodium CSP junction region variant comprises the amino acid sequence of AAKQ. [0067] In some embodiments, one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise one or more Plasmodium CSP N-terminal end regions or portions thereof.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise two or more Plasmodium CSP N-terminal end regions or portions thereof.
  • each Plasmodium CSP N-terminal end region consists of an amino acid sequence according to SEQ ID NO: 285.
  • a second polypeptide does not comprise a Plasmodium CSP N-terminal end region or any portion thereof.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise one or more Plasmodium CSP N-terminal regions or portions thereof.
  • each Plasmodium CSP N-terminal region comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 288.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise an antigenic portion of a Plasmodium CSP N-terminal region.
  • an antigenic portion of a Plasmodium CSP N-terminal region is a Plasmodium CSP N-terminal start region.
  • a Plasmodium CSP N-terminal start region comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 1010.
  • a second polypeptide does not comprise a Plasmodium CSP N-terminal region or any portion thereof.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise one or more Plasmodium CSP major repeat regions or portions thereof.
  • one or more Plasmodium CSP major repeat regions or portions thereof comprise the amino acid sequence NANPNA or NPNANP. In some embodiments, one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise at least two Plasmodium CSP major repeat region portions. In some embodiments, one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise at least two Plasmodium CSP major repeat region portions, wherein each CSP major repeat region portion comprises at least 4 and at most 7 repeats of the sequence NANP.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise two or three Plasmodium CSP major repeat region portions, wherein each CSP major repeat region portion comprises 6 repeats of the sequence NANP (SEQ ID NO: 230). In some embodiments, one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise two or three Plasmodium CSP major repeat region portions, wherein each CSP major repeat region portion comprises 6 repeats of the sequence NANP (SEQ ID NO: 230).
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise exactly one Plasmodium CSP major repeat region or portion thereof, and the Plasmodium CSP major repeat region or portion thereof comprises a total of at least 2 and at most 35 repeats of the amino acid sequence NANP (SEQ ID NO: 230).
  • a Plasmodium CSP major repeat region or portion thereof comprises two contiguous stretches of repeats of the amino acid sequence NANP (SEQ ID NO: 230), and wherein the two contiguous stretches of repeats of the amino acid sequence NANP (SEQ ID NO: 230) flank an amino acid sequence of NVDP (SEQ ID NO:229).
  • a Plasmodium CSP major repeat region comprises, in N-terminus to C-terminus order, 17 repeats of the amino acid sequence NANP (SEQ ID NO: 230), an amino acid sequence of NVDP (SEQ ID NO:229), and 18 repeats of the amino acid sequence NANP (SEQ ID NO: 230).
  • a portion of a Plasmodium CSP major repeat region consists of at most 18 contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230).
  • a portion of the Plasmodium CSP major repeat region consists of 2 contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230).
  • a Plasmodium CSP major repeat region comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 305.
  • one or more Plasmodium CSP polypeptide regions or antigenic portions thereof comprises an antigenic portion of a Plasmodium CSP major repeat region.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises at least six repeats of the amino acid sequence of NANP.
  • an antigenic portion of a Plasmodium CSP major repeat region further comprises an asparagine-alanine positioned immediately following of the six repeats of the amino acid sequence of NANP (SEQ ID NO: 230).
  • an antigenic portion of a Plasmodium CSP major repeat region comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 303.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises eighteen repeats of the amino acid sequence of NANP (SEQ ID NO: 230). In some embodiments, an antigenic portion of a Plasmodium CSP major repeat region comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 308. [0079] In some embodiments, a second polypeptide does not comprise a Plasmodium CSP N-terminal region or any portion thereof.
  • a second polypeptide does not comprise a Plasmodium CSP C-terminal region or any portion thereof. In some embodiments, a second polypeptide does not comprise (i) a Plasmodium CSP N-terminal region or any portion thereof and/or (ii) a Plasmodium CSP C-terminal region or any portion thereof. [0080] In some embodiments, a second polypeptide does not comprise a Plasmodium CSP major repeat region or a portion of a Plasmodium CSP major repeat region comprising the amino acid sequence NPNA (SEQ ID NO: 228).
  • one or more Plasmodium CSP polypeptide regions or portions thereof, if present in a second polypeptide, are in the following N-terminus to C-terminus order: (i) one or more Plasmodium CSP N- terminal regions or portions thereof; (ii) one or more Plasmodium CSP N-terminal end regions or portions thereof; (iii) one or more Plasmodium CSP junction regions, portions thereof, or variants thereof; (iv) one or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (v) one or more Plasmodium CSP major repeat regions or portions thereof; and (vi) one or more Plasmodium CSP C-terminal regions or portions thereof.
  • one or more Plasmodium CSP polypeptide regions or portions thereof, if present in the second polypeptide are in the following N-terminus to C-terminus order: (i) one Plasmodium CSP N-terminal region or portion thereof; (ii) one Plasmodium CSP N-terminal end region or portion thereof; (iii) one Plasmodium CSP junction region, portion thereof, or variant thereof; (iv) one or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (v) one Plasmodium CSP major repeat region or portion thereof; and (vi) one Plasmodium CSP C-terminal region or portion thereof.
  • a second polypeptide comprises one or more helper antigens.
  • one or more helper antigens comprise a Plasmodium antigen.
  • one or more helper antigens are Plasmodium 2-phospho-D-glycerate hydro-lyase antigen, Plasmodium liver stage antigen 1(a), (LSA-1(a)), Plasmodium liver stage antigen 1(b) (LSA-1(b)), Plasmodium thrombospondin-related anonymous protein (TRAP), Plasmodium liver stage associated protein 1 (LSAP1), Plasmodium liver stage associated protein 2 (LSAP2), Plasmodium UIS3, Plasmodium ETRAMP10.3, Plasmodium liver specific protein 1 (LISP-1), Plasmodium liver specific protein 2 (LISP-2), Plasmodium liver stage antigen 3 (LSA-3), Plasmodium EXP1, Plasmodium E140, Plasmodium reticulocyte-binding protein homolog
  • one or more helper antigens comprise or consist of a P. falciparum 2-phospho- D-glycerate hydro-lyase antigen.
  • a P. falciparum 2-phospho-D-glycerate hydro-lyase antigen comprises or consists of an amino acid sequence according to SEQ ID NO: 388.
  • one or more helper antigens comprise or consist of a P. falciparum liver-stage antigen 3.
  • a P. falciparum liver-stage antigen 3 comprises or consists of an amino acid sequence according to SEQ ID NO: 391.
  • one or more helper antigens comprise an Anopheles antigen.
  • a helper antigen comprises or consists of an Anopheles gambiae TRIO.
  • an Anopheles gambiae TRIO comprises or consists of an amino acid sequence according to SEQ ID NO: 393.
  • a second polypeptide comprises a secretory signal and the helper antigen immediately follows the secretory signal.
  • a second polypeptide comprises a helper antigen at the C-terminus of the second polypeptide.
  • a second polypeptide comprises a multimerization region.
  • a multimerization region comprises or consists of a trimerization region.
  • a trimerization region comprises or consists of a fibritin region.
  • a fibritin region comprises or consists of an amino acid sequence according to SEQ ID NO: 399.
  • a second polypeptide comprises a multimerization region at the N-terminus of the second polypeptide.
  • a second polypeptide comprises a self-aggregation region.
  • a self-aggregation region comprises or consists of a ferritin region.
  • a ferritin region comprises or consists of an amino acid sequence according to SEQ ID NO: 402.
  • a polypeptide comprises a self-aggregation region at the N-terminus of the polypeptide.
  • a second polypeptide comprises a secretory signal.
  • a secretory signal comprises or consists of a Plasmodium secretory signal.
  • a Plasmodium secretory signal comprises or consists of a Plasmodium CSP secretory signal.
  • a Plasmodium CSP secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 332.
  • a secretory signal comprises or consists of a heterologous secretory signal.
  • a heterologous secretory signal comprises or consists of a non-human secretory signal. In some embodiments, a heterologous secretory signal comprises or consists of a viral secretory signal. In some embodiments, a viral secretory signal comprises or consists of an HSV secretory signal. In some embodiments, a HSV secretory signal comprises or consists of an HSV-1 or HSV-2 secretory signal. In some embodiments, a HSV secretory signal comprises or consists of an HSV glycoprotein D (gD) secretory signal. In some embodiments, a HSV gD secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 314.
  • a HSV gD secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 320.
  • a viral secretory signal comprises or consists of an Ebola virus secretory signal.
  • a Ebola virus secretory signal comprises or consists of an Ebola virus spike glycoprotein (SGP) secretory signal.
  • a Ebola virus SGP secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 335.
  • a secretory signal is located at the N-terminus of a second polypeptide. In some embodiments, a second polypeptide does not comprise a secretory signal.
  • a second polypeptide comprises a transmembrane region.
  • a transmembrane region comprises or consists of a Plasmodium transmembrane region.
  • a Plasmodium transmembrane region comprises or consists of a Plasmodium CSP glycosylphosphatidylinositol (GPI) anchor region.
  • a Plasmodium CSP GPI anchor region comprises or consists of an amino acid sequence according to SEQ ID NO: 385.
  • a transmembrane region comprises or consists of a heterologous transmembrane region.
  • a heterologous transmembrane region does not comprise a hemagglutinin transmembrane region.
  • a heterologous transmembrane region comprises or consists of a non-human transmembrane region.
  • a heterologous transmembrane region comprises or consists of a viral transmembrane region.
  • a viral transmembrane region comprises or consists of an HSV transmembrane region.
  • a HSV transmembrane region comprises or consists of an HSV-1 or HSV-2 transmembrane region.
  • a HSV transmembrane region comprises or consists of an HSV gD transmembrane region. In some embodiments, a HSV gD transmembrane region comprises or consists of an amino acid sequence according to SEQ ID NO: 379. In some embodiments, a heterologous transmembrane region comprises or consists of a human transmembrane region. In some embodiments, a human transmembrane region comprises or consists of a human decay accelerating factor glycosylphosphatidylinositol (hDAF-GPI) anchor region. In some embodiments, a hDAF-GPI anchor region comprises or consists of an amino acid sequence according to SEQ ID NO: 382.
  • a second polypeptide does not comprise a transmembrane region.
  • a second polypeptide comprises one or more linkers.
  • one or more linkers comprise one or more glycine-serine linkers.
  • one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 404.
  • one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 411.
  • one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 408.
  • one or more linkers comprise or consist of an amino acid sequence according to SEQ ID NO: 412.
  • a second polypeptide comprises a linker between the C-terminal region or portion thereof and the transmembrane region. In some embodiments, a second polypeptide comprises a linker after an amino acid sequence of NANPNVDP (SEQ ID NO: 223). In some embodiments, a second polypeptide comprises a secretory signal. In some embodiments, a second polypeptide comprises one or more Plasmodium CSP junction regions, portions thereof, or variants thereof. In some embodiments, a second polypeptide comprises one or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223). In some embodiments, a second polypeptide comprises one or more Plasmodium CSP C-terminal regions or portions thereof.
  • a second polypeptide comprises a transmembrane region. In some embodiments, a second polypeptide does not comprise an amino acid sequence of NPNA (SEQ ID NO: 228). In some embodiments, a second polypeptide does not comprise a Plasmodium CSP N-terminal region or portion thereof.
  • a second polypeptide comprises: (i) a secretory signal; (ii) one or more Plasmodium CSP junction regions, portions thereof, or variants thereof; (iii) one or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) one or more Plasmodium CSP C- terminal regions or portions thereof; and (v) a transmembrane region, and a second polypeptide does not comprise: (a) an amino acid sequence of NPNA (SEQ ID NO: 228); and (b) a Plasmodium CSP N-terminal region or portion thereof. In some embodiments, a second polypeptide does not comprise a Plasmodium CSP N-terminal end region.
  • a second polypeptide comprises one or more Plasmodium CSP N-terminal end regions or portions thereof.
  • a second polypeptide comprises: (i) a secretory signal; (ii) one or more Plasmodium CSP junction regions, portions thereof, or variants thereof; (iii) one or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); and (iv) one or more Plasmodium CSP C-terminal regions or portions thereof.
  • a second polypeptide comprises a secretory signal.
  • a second polypeptide comprises three or more Plasmodium CSP junction regions, portions thereof, or variants thereof.
  • a second polypeptide comprises three or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223). In some embodiments, a second polypeptide comprises two or more Plasmodium CSP major repeat region portions. In some embodiments, a second polypeptide does not comprise a Plasmodium CSP N- terminal region or portion thereof. In some embodiments, a second polypeptide does not comprise a Plasmodium CSP C-terminal region or portion thereof.
  • a second polypeptide comprises: (i) a secretory signal; (ii) three or more Plasmodium CSP junction regions, portions thereof, or variants thereof; (iii) three or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); and (iv) two or more Plasmodium CSP major repeat region portions, and a second polypeptide does not comprise: (a) a Plasmodium CSP N-terminal region or portion thereof; and (b) a Plasmodium CSP C-terminal region or portion thereof.
  • a second polypeptide comprises an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to a sequence of SEQ ID NO: 108.
  • a second polypeptide comprises: (i) a secretory signal; (ii) one or more Plasmodium CSP junction regions, portions thereof, or variants thereof; (iii) one or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) one or more Plasmodium CSP C-terminal regions or portions thereof; and (v) a transmembrane region.
  • a second polypeptide comprises: (i) a secretory signal; (ii) one or more Plasmodium CSP junction regions, portions thereof, or variants thereof; (iii) one or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) one or more Plasmodium CSP C-terminal regions or portions thereof; and (v) a transmembrane region.
  • a second polypeptide comprises: (i) a secretory signal; (ii) three or more Plasmodium CSP junction regions, portions thereof, or variants thereof; (iii) three or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) two or more Plasmodium CSP major repeat region portions; and (v) a transmembrane region, and a second polypeptide does not comprise: (a) a Plasmodium CSP N- terminal region or portion thereof; and (b) a Plasmodium CSP C-terminal region or portion thereof.
  • a second polypeptide comprises an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to a sequence of SEQ ID NO: 107 or 109.
  • a second polypeptide comprises: (i) a secretory signal; (ii) three or more Plasmodium CSP junction regions, portions thereof, or variants thereof; (iii) three or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); and (iv) two or more Plasmodium CSP major repeat region portions, wherein a second polypeptide does not comprise: (a) a Plasmodium CSP N-terminal region or portion thereof; and (b) a Plasmodium CSP C-terminal region or portion thereof.
  • a second polypeptide comprises an amino acid sequence that has at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a sequence of SEQ ID NO: 110 or 111.
  • a second polypeptide comprises one or more helper antigens.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal end region; (iii) a Plasmodium CSP junction region; (iv) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (v) a Plasmodium CSP C-terminal region; and (vi) five antigenic repeat regions, wherein each antigenic repeat region comprises: (A) a linker; and (B) a helper antigen, and a second polypeptide does not comprise any of: (a) an amino acid sequence of NPNA (SEQ ID NO: 228); (b) a Plasmodium CSP N-terminal region or portion thereof; and (c) a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 36.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a helper antigen; (iii) a linker; (iv) a Plasmodium CSP N-terminal end region; (v) a Plasmodium CSP junction region; (vi) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (vii) a Plasmodium CSP C-terminal region; (viii) a serine- valine sequence immediately following the Plasmodium CSP C-terminal region; (ix) a linker; and (x) a transmembrane region, and a second polypeptide does not comprise any of: (a) an amino acid sequence of NPNA (SEQ ID NO: 228); and (b) a Plasmodium CSP N-terminal region or portion thereof.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 39.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a portion of a Plasmodium CSP junction region; (iii) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) a Plasmodium CSP C-terminal region; (v) a serine-valine sequence immediately following the Plasmodium CSP C- terminal region; (vi) a linker; and (vii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; (b) a Plasmodium CSP N-terminal end region or portion thereof; and (c) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 57.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a portion of a Plasmodium CSP junction region; (iii) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) a Plasmodium CSP C-terminal region; (v) a serine-valine sequence immediately following the Plasmodium CSP C- terminal region; (vi) a linker; and (vii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; (b) a Plasmodium CSP N-terminal end region or portion thereof; (c) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 60.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP junction region; (iii) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) a Plasmodium CSP C-terminal region; (v) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region; (vi) a linker; and (vii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; (b) a Plasmodium CSP N-terminal end region or portion thereof; and (c) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 63.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP junction region; (iii) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) a Plasmodium CSP C-terminal region; (v) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region; (vi) a linker; and (vii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; (b) a Plasmodium CSP N-terminal end region or portion thereof; and (c) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 66.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP junction region variant; (iii) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) a Plasmodium CSP C-terminal region; (v) a serine-valine sequence immediately following the Plasmodium CSP C- terminal region; (vi) a linker; and (vii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; (b) a Plasmodium CSP N-terminal end region or portion thereof; and (c) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 69.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP junction region variant; (iii) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) a Plasmodium CSP C-terminal region; (v) a serine-valine sequence immediately following the Plasmodium CSP C- terminal region; (vi) a linker; and (vii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; (b) a Plasmodium CSP N-terminal end region or portion thereof; and (c) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 72.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a portion of a Plasmodium CSP junction region; (iii) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) a Plasmodium CSP C-terminal region; (v) a serine-valine sequence immediately following the Plasmodium CSP C- terminal region; (vi) a linker; and (vii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; (b) a Plasmodium CSP N-terminal end region or portion thereof; and (c) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 75.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a portion of a Plasmodium CSP junction region; (iii) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) a Plasmodium CSP C-terminal region; (v) a serine-valine sequence immediately following the Plasmodium CSP C- terminal region; (vi) a linker; and (vii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; (b) a Plasmodium CSP N-terminal end region or portion thereof; and (c) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 78.
  • second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal end region; (iii) a Plasmodium CSP junction region; (iv) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (v) a Plasmodium CSP C-terminal region; (vi) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region; (vii) a linker; and (viii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; (b) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 81.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal end region; (iii) a Plasmodium CSP junction region variant; (iv) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (v) a Plasmodium CSP C-terminal region; (vi) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region; (vii) a linker; and (viii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; and (b) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 84.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal end region; (iii) a Plasmodium CSP junction region; (iv) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (v) a Plasmodium CSP C-terminal region; and (vi) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; and (b) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 102.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal end region; (iii) a Plasmodium CSP junction region; (iv) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (v) a Plasmodium CSP C-terminal region; and (vi) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; (b) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 105.
  • a second polypeptide comprises: (i) a secretory signal; (ii) two or more Plasmodium CSP neutralizing region repeats, wherein each Plasmodium CSP neutralizing region repeat comprises or consists of: (a) a Plasmodium CSP N-terminal end region; (b) a Plasmodium CSP junction region; (c) two repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); and (d) a linker; (iii) a portion of a Plasmodium CSP major repeat region; (iv) a Plasmodium CSP C-terminal region; (v) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region; (vi) a linker; and (vii) a transmembrane region, and a second polypeptide does not comprise a Plasmodium CSP N-terminal region or portion thereof.
  • a second polypeptide comprises exactly four Plasmodium CSP neutralizing region repeats.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 87.
  • a second polypeptide comprises: (i) a secretory signal; (ii) one Plasmodium CSP junction region; (iii) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (iv) a Plasmodium CSP major repeat region; (v) one Plasmodium CSP C-terminal region; (vi) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region; (vii) a linker; and (viii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; and (b) a Plasmodium CSP N-terminal end region or portion thereof.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 30.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a portion of a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; and (viii) a serine immediately following the Plasmodium CSP C- terminal region, and a second polypeptide does not comprise a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 27.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; and (viii) a serine immediately following the Plasmodium CSP C-terminal region, and a second polypeptide does not comprise a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 8.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; and (viii) a serine immediately following the Plasmodium CSP C-terminal region, and a second polypeptide does not comprise a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 24.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; and (viii) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region, and a second polypeptide does not comprise a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 99.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; and (viii) a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 33.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; (viii) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region; (ix) a linker; and (x) a multimerization region, and a second polypeptide does not comprise a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 42.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; (viii) a serine immediately following the Plasmodium CSP C-terminal region; (ix) a linker; and (x) a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 48.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; (viii) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region; (ix) a linker; and (x) a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 90.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; (viii) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region; (ix) a linker; and (x) a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 90.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; (viii) a serine immediately following the Plasmodium CSP C-terminal region; and (ix) a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 21.
  • a second polypeptide comprises: (i) a secretory signal; (ii) an antigenic portion of a Plasmodium CSP major repeat region; (iii) a Plasmodium CSP C-terminal region; and (iv) a serine-valine immediately following the Plasmodium CSP C-terminal region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 51.
  • a second polypeptide comprises: (i) a secretory signal; (ii) an antigenic portion of a Plasmodium CSP major repeat region; (iii) a Plasmodium CSP C-terminal region; (iv) a serine immediately following the Plasmodium CSP C-terminal region; (v) a linker; and (vi) a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 54.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise in N-terminus to C-terminus order: (i) three contiguous repeats of the amino acid sequence NANPNVDP (SEQ ID NO: 223); (ii) six contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230); (iii) three contiguous repeats of the amino acid sequence NANPNVDP (SEQ ID NO: 223); (iv) six contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230); and (v) three contiguous repeats of the amino acid sequence NANPNVDP (SEQ ID NO: 223).
  • a second polypeptide further comprises: (i) one or more Plasmodium CSP N-terminal regions or portions thereof; (ii) one or more Plasmodium CSP N-terminal end regions or portions thereof; (iii) one or more Plasmodium CSP junction regions, portions thereof, or variants thereof; (iv) one or more Plasmodium CSP C-terminal regions or portions thereof; or (v) a combination thereof.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in a second polypeptide comprise in N-terminus to C-terminus order: (i) a Plasmodium CSP N-terminal end region or portion thereof; (ii) a Plasmodium CSP R1 region or portion thereof; (iii) a Plasmodium CSP junction region or portion thereof; (iv) three contiguous repeats of the amino acid sequence NANPNVDP (SEQ ID NO: 223); (v) six contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230); (vi) three contiguous repeats of the amino acid sequence NANPNVDP (SEQ ID NO: 223); (vii) six contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230); (viii) three contiguous repeats of the amino acid sequence NANPNVDP (SEQ ID NO: 223); and (ix) a Plasmodium CSP C-terminal
  • a second polypeptide further comprises a Plasmodium CSP GPI domain.
  • a Plasmodium CSP C-terminal region or portion thereof comprised in the second polypeptide comprises a substitution at a fucosylation site.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in the second polypeptide comprise in N-terminus to C-terminus order three repeating domains.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in the second polypeptide comprise in N-terminus to C-terminus order three repeating domains, wherein each repeating domain comprises one or more Plasmodium CSP junction regions, portions thereof, or variants thereof.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in the second polypeptide comprise in N-terminus to C-terminus order three repeating domains, wherein each repeating domain comprises three contiguous repeats of the amino acid sequence NANPNVDP (SEQ ID NO: 223).
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in the second polypeptide comprise in N-terminus to C-terminus order three repeating domains, wherein each repeating domain comprises six contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230).
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in the second polypeptide comprise in N- terminus to C-terminus order three repeating domains, wherein each repeating domain comprises: (i) one or more Plasmodium CSP junction regions, portions thereof, or variants thereof; (ii) three contiguous repeats of the amino acid sequence NANPNVDP (SEQ ID NO: 223); and (iii) six contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230).
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in the second polypeptide comprise in N-terminus to C-terminus order three repeating domains, wherein each repeating domain comprises one or more Plasmodium R1 regions or portions thereof.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in the second polypeptide comprise in N- terminus to C-terminus order three repeating domains, wherein each repeating domain comprises one or more Plasmodium CSP junction regions, portions thereof, or variants thereof.
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in the second polypeptide comprise in N-terminus to C-terminus order three repeating domains, wherein each repeating domain comprises three contiguous repeats of the amino acid sequence NANPNVDP (SEQ ID NO: 223).
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in the second polypeptide comprise in N-terminus to C-terminus order three repeating domains, wherein each repeating domain comprises six contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230).
  • one or more Plasmodium CSP polypeptide regions or portions thereof comprised in the second polypeptide comprise in N-terminus to C-terminus order three repeating domains, wherein each repeating domain comprises: (i) one or more Plasmodium R1 regions or portions thereof; (ii) one or more Plasmodium CSP junction regions, portions thereof, or variants thereof; (iii) three contiguous repeats of the amino acid sequence NANPNVDP (SEQ ID NO: 223); and (iv) six contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230).
  • a combination further comprises one or more linker sequences.
  • a linker is a glycine-serine linker.
  • a second polypeptide comprises a linker sequence following each six contiguous repeats of the amino acid sequence NANP. [0134] In some embodiments, a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junctional region; (v) a Plasmodium CSP minor repeat region; (vi) an antigenic portion of a Plasmodium CSP major repeat region; and (vii) a Plasmodium CSP C-terminal region.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises or consists of eighteen repeats of the amino acid sequence of NANP.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence according to SEQ ID NO: 117.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises of six repeats of the amino acid sequence of NANP (SEQ ID NO: 230).
  • an antigenic portion of a Plasmodium CSP major repeat region further comprises an asparagine-alanine positioned immediately following the six repeats of the amino acid sequence of NANP (SEQ ID NO: 230).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence according to SEQ ID NO: 122.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junctional region; (v) a Plasmodium CSP minor repeat region; (vi) an antigenic portion of a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; and (viii) a transmembrane region.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises or consists of eighteen repeats of the amino acid sequence of NANP (SEQ ID NO: 230).
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence according to SEQ ID NO: 112.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a linker; (v) a Plasmodium CSP junctional region; (vi) a Plasmodium CSP minor repeat region; (vii) an antigenic portion of a Plasmodium CSP major repeat region; (viii) a Plasmodium CSP C-terminal region; (ix) a linker; and (x) a transmembrane region.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises or consists of eighteen repeats of the amino acid sequence of NANP.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence according to SEQ ID NO: 125.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junctional region; (v) a Plasmodium CSP minor repeat region; (vi) a Plasmodium CSP major repeat region; (vii) a linker; and (viii) a transmembrane region.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence according to SEQ ID NO: 130.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP junctional region; (iii) a Plasmodium CSP minor repeat region; (iv) an antigenic portion of a Plasmodium CSP major repeat region; (v) a linker; (vi) an antigenic portion of a Plasmodium CSP C-terminal region; (vii) a serine-valine; (viii) a linker; and (ix) a multimerization region.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises of six repeats of the amino acid sequence of NANP.
  • an antigenic portion of a Plasmodium CSP major repeat region further comprises an asparagine-alanine positioned immediately following the six repeats of the amino acid sequence of NANP (SEQ ID NO: 230).
  • an antigenic portion of a Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 259.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence according to SEQ ID NO: 135.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP junctional region; (iii) a Plasmodium CSP minor repeat region; (iv) an antigenic portion of a Plasmodium CSP major repeat region; (v) a linker; (vi) an antigenic portion of a Plasmodium CSP C-terminal region; (vii) a serine-valine; (viii) a linker; and (ix) a self-aggregation region.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises of six repeats of the amino acid sequence of NANP (SEQ ID NO: 230).
  • an antigenic portion of a Plasmodium CSP major repeat region further comprises an asparagine- alanine positioned immediately following the six repeats of the amino acid sequence of NANP (SEQ ID NO: 230).
  • an antigenic portion of a Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, at least 90%, at least 95%, at least 98%, or 100% identical to the amino acid sequence of SEQ ID NO: 259.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence according to SEQ ID NO: 138.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP junctional region; (iii) a Plasmodium CSP minor repeat region; (iv) an antigenic portion of a Plasmodium CSP major repeat region; (v) a linker; (vi) an antigenic portion of a Plasmodium CSP C-terminal region; (vii) a serine-valine; (viii) a linker; and (ix) a transmembrane region.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises of six repeats of the amino acid sequence of NANP (SEQ ID NO: 230).
  • an antigenic portion of a Plasmodium CSP major repeat region further comprises an asparagine- alanine positioned immediately following the six repeats of the amino acid sequence of NANP (SEQ ID NO: 230).
  • an antigenic portion of a Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%, at least 90%, at least 95%, at least 98%, or 100% identical to the amino acid sequence of SEQ ID NO: 259.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the amino acid sequence according to SEQ ID NO: 141.
  • a second polypeptide comprises: (i) a secretory signal; (ii) a Plasmodium CSP N- terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junctional region; (v) a Plasmodium CSP minor repeat region; (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region variant; and (viii) a transmembrane region.
  • a secretory signal comprises or consists of a Plasmodium secretory signal.
  • a Plasmodium secretory signal comprises or consists of a Plasmodium CSP secretory signal. In some embodiments, a Plasmodium CSP secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 332. In some embodiments, a transmembrane region comprises or consists of a Plasmodium transmembrane region. In some embodiments, a Plasmodium transmembrane region comprises or consists of a Plasmodium CSP glycosylphosphatidylinositol (GPI) anchor region. In some embodiments, a Plasmodium CSP GPI anchor region comprises or consists of an amino acid sequence according to SEQ ID NO: 385.
  • a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to the amino acid sequence according to SEQ ID NO: 989. In some embodiments, a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 989. In some embodiments, a polyribonucleotide comprises or consists of a nucleic acid sequence with at least 85% sequence identity to the nucleic acid sequence according to SEQ ID NO: 991. In some embodiments, a polyribonucleotide comprises or consists of a nucleic acid sequence according to SEQ ID NO: 991.
  • a second polypeptide comprises: (i) a secretory signal; (ii) an antigenic portion of a Plasmodium CSP N-terminal region; (iii) a first linker; (iv) a Plasmodium CSP N-terminal end region; (v) a Plasmodium CSP junctional region; (vi) a Plasmodium CSP minor repeat region; (vii) an antigenic portion of a Plasmodium CSP major repeat region; (viii) a Plasmodium CSP C-terminal region; (ix) a serine-valine sequence; (x) a second linker; and (xi) a transmembrane region.
  • a secretory signal comprises or consists of a Plasmodium secretory signal.
  • a Plasmodium secretory signal comprises or consists of a Plasmodium CSP secretory signal.
  • a Plasmodium CSP secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 332.
  • a transmembrane region comprises or consists of an HSV transmembrane region.
  • a HSV transmembrane region comprises or consists of an HSV-1 or HSV-2 transmembrane region.
  • a HSV transmembrane region comprises or consists of an HSV gD transmembrane region.
  • a HSV gD transmembrane region comprises or consists of an amino acid sequence according to SEQ ID NO: 379.
  • an antigenic portion of a Plasmodium CSP N-terminal region comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 98%, or 100% identical to the amino acid sequence of according to SEQ ID NO: 1010.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises or consists of eighteen repeats of the amino acid sequence of NANP.
  • a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to the amino acid sequence according to SEQ ID NO: 997.
  • a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 997.
  • a polyribonucleotide comprises or consists of a nucleic acid sequence with at least 85% sequence identity to the nucleic acid sequence according to SEQ ID NO: 999.
  • a polyribonucleotide comprises or consists of a nucleic acid sequence according to SEQ ID NO: 999.
  • second polypeptide comprises: (i) a secretory signal; (ii) an antigenic portion of a Plasmodium CSP N-terminal region; (iii) a first linker; (iv) a Plasmodium CSP N-terminal end region; (v) a Plasmodium CSP junctional region; (vi) a Plasmodium CSP minor repeat region; (vii) an antigenic portion of a Plasmodium CSP major repeat region; (viii) a second linker; (ix) an antigenic portion of a Plasmodium CSP C-terminal region; (x) a serine-valine sequence; (xi) a third linker; and (xii) a transmembrane region.
  • a secretory signal comprises or consists of a Plasmodium secretory signal.
  • a Plasmodium secretory signal comprises or consists of a Plasmodium CSP secretory signal.
  • a Plasmodium CSP secretory signal comprises or consists of an amino acid sequence according to SEQ ID NO: 332.
  • a transmembrane region comprises or consists of an HSV transmembrane region.
  • a HSV transmembrane region comprises or consists of an HSV-1 or HSV-2 transmembrane region.
  • a HSV transmembrane region comprises or consists of an HSV gD transmembrane region.
  • a HSV gD transmembrane region comprises or consists of an amino acid sequence according to SEQ ID NO: 379.
  • an antigenic portion of a Plasmodium CSP N-terminal region comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 98%, or 100% identical to the amino acid sequence of according to SEQ ID NO: 1010.
  • an antigenic portion of a Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 98%, or 100% identical to the amino acid sequence of SEQ ID NO: 259.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises or consists of eighteen repeats of the amino acid sequence of NANP.
  • a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to the amino acid sequence according to SEQ ID NO: 1000.
  • a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 1000.
  • a polyribonucleotide comprises or consists of a nucleic acid sequence with at least 85% sequence identity to the nucleic acid sequence according to SEQ ID NO: 1002.
  • a polyribonucleotide comprises or consists of a nucleic acid sequence according to SEQ ID NO: 1002.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises six repeats of the amino acid sequence of NANP.
  • an antigenic portion of a Plasmodium CSP major repeat region further comprises an asparagine-alanine positioned immediately following the six repeats of the amino acid sequence of NANP.
  • a polypeptide comprises or consists of an amino acid sequence with at least 85% sequence identity to the amino acid sequence according to SEQ ID NO: 1003.
  • a polypeptide comprises or consists of an amino acid sequence according to SEQ ID NO: 1003.
  • a polyribonucleotide comprises or consists of a nucleic acid sequence with at least 85% sequence identity to the nucleic acid sequence according to SEQ ID NO: 1005. In some embodiments, a polyribonucleotide comprises or consists of a nucleic acid sequence according to SEQ ID NO: 1005. In some embodiments, when present, the features are in a second polypeptide in numerical order from the C-terminus to the N-terminus.
  • a combination comprises a first polypeptide that comprises: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; and (v) an antigenic Plasmodium LSAP2 polypeptide fragment.
  • a combination comprises a second polypeptide that comprises: (i) a secretory signal; (ii) a Plasmodium CSP N-terminal region; (iii) a Plasmodium CSP N- terminal end region; (iv) a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; and (viii) a transmembrane region.
  • a combination comprises a first polypeptide that comprises: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; and (v) an antigenic Plasmodium LSAP2 polypeptide fragment, and a second polypeptide that comprises: (i) a secretory signal; (ii) a Plasmodium CSP N-terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; and (viii) a transmembra first polypeptid
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 33.
  • a third polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 209.
  • a combination comprises a first polypeptide that comprises: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; and (v) an antigenic Plasmodium LSAP2 polypeptide fragment.
  • a combination comprises a second polypeptide that comprises: (i) a secretory signal; (ii) a Plasmodium CSP N-terminal end region; (iii) a Plasmodium CSP junction region; (iv) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (v) a Plasmodium CSP C-terminal region; (vi) a serine-valine sequence immediately following the Plasmodium CSP C- terminal region; (vii) a linker; and (viii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; and (b) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a first polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 203.
  • a combination comprises a first polypeptide that comprises: (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (iii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a combination comprises a first polypeptide that comprises: (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (iii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment, and a second polypeptide that comprises: (i) a secretory signal; (ii) a Plasmodium CSP N-terminal region; (iii) a Plasmodium CSP N-terminal end region; (iv) a Plasmodium CSP junction region; (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (vi) a Plasmodium CSP major repeat region; (vii) a Plasmodium CSP C-terminal region; and (viii) a transmembrane region.
  • a first polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 209.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 33.
  • a combination comprises a first polypeptide that comprises: (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (iii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a combination comprises a second polypeptide that comprises: (i) a secretory signal; (ii) a Plasmodium CSP N-terminal end region; (iii) a Plasmodium CSP junction region; (iv) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (v) a Plasmodium CSP C-terminal region; (vi) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region; (vii) a linker; and (viii) a transmembrane region, and a second polypeptide does not comprise any of: (a) a Plasmodium CSP N-terminal region or portion thereof; and (b) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a combination comprises a first polypeptide that comprises: (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (iii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment, and a second polypeptide that comprises: (i) a secretory signal; (ii) a Plasmodium CSP N-terminal end region; (iii) a Plasmodium CSP junction region; (iv) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223); (v) a Plasmodium CSP C-terminal region; (vi) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region; (vii) a linker; and (viii) a transmembrane region, and a second polypeptide does
  • a first polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 209.
  • a second polypeptide comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 81.
  • Plasmodium is Plasmodium falciparum.
  • one or more Plasmodium CSP polypeptide regions or portions thereof are one or more P. falciparum CSP polypeptide regions or portions thereof.
  • one or more Plasmodium T-cell antigens are one or more P.
  • a first polyribonucleotide and/or second polyribonucleotide is an isolated polyribonucleotide.
  • a third polyribonucleotide is an isolated polyribonucleotide.
  • a first polyribonucleotide and/or second polyribonucleotide is an engineered polyribonucleotide.
  • a third polyribonucleotide is an engineered polyribonucleotide.
  • a first polyribonucleotide and/or second polyribonucleotide is a codon-optimized polyribonucleotide.
  • a third polyribonucleotide is a codon-optimized polyribonucleotide.
  • the present disclosure further provides RNA constructs.
  • a first polyribonucleotide is comprised in a first RNA construct, wherein the first RNA construct comprises in 5' to 3' order: (i) a 5' UTR; (ii) the first polyribonucleotide; (iii) a 3' UTR; and (iv) a polyA tail sequence.
  • second polyribonucleotide is comprised in a second RNA construct, wherein the second RNA construct comprises in 5' to 3' order: (i) a 5' UTR; (ii) the second polyribonucleotide; (iii) a 3' UTR; and (iv) a polyA tail sequence.
  • a 5' UTR of a first and/or second RNA construct comprises or consists of a modified human alpha-globin 5'-UTR.
  • a 5' UTR of a first and/or second RNA construct consists of a ribonucleic acid sequence according to SEQ ID NO: 565.
  • a 3' UTR of a first and/or second RNA construct comprises or consists of a first sequence from the amino terminal enhancer of split (AES) messenger RNA and a second sequence from the mitochondrial encoded 12S ribosomal RNA.
  • a 3' UTR of a first and/or second RNA construct consists of a ribonucleic acid sequence according to SEQ ID NO: 567.
  • a 5' UTR of a first and/or second RNA construct comprises or consists of a modified human alpha-globin 5'-UTR and a 3' UTR of a first and/or second RNA construct comprises or consists of a first sequence from the amino terminal enhancer of split (AES) messenger RNA and a second sequence from the mitochondrial encoded 12S ribosomal RNA.
  • AES amino terminal enhancer of split
  • a polyA tail sequence of a first and/or second RNA construct is a split polyA tail sequence.
  • a split polyA tail sequence consists of a ribonucleic acid sequence according to SEQ ID NO: 569.
  • a first and/or second RNA construct further comprise a 5' cap.
  • a first and/or second RNA construct comprise a cap proximal sequence comprising positions +1, +2, +3, +4, and +5 of the polyribonucleotide.
  • a 5' cap comprises or consists of m7(3’OMeG)(5')ppp(5')(2'OMeA1)pG2, wherein A1 is position +1 of the polyribonucleotide, and G2 is position +2 of the polyribonucleotide.
  • a cap proximal sequence comprises A 1 and G 2 of the Cap1 structure, and a sequence comprising: A 3 A 4 U 5 (SEQ ID NO: 571) at positions +3, +4 and +5 respectively of the polyribonucleotide.
  • a first and/or second RNA construct includes modified uridines in place of all uridines.
  • modified uridines are each N1-methyl-pseudouridine.
  • a first and/or second pharmaceutical composition further comprises lipid nanoparticles, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes.
  • a first and/or second polyribonucleotide is fully or partially encapsulated within the lipid nanoparticles, polyplexes (PLX), lipidated polyplexes (LPLX), or liposomes.
  • a first and/or second pharmaceutical composition further comprises lipid nanoparticles.
  • a first polyribonucleotide is encapsulated within the lipid nanoparticles.
  • a second polyribonucleotide is encapsulated within the lipid nanoparticles.
  • a first and/or second pharmaceutical composition comprises at least one pharmaceutically acceptable excipient.
  • a combination is for use in the treatment of a malaria infection. In some embodiments, a combination is for use in the prevention of a malaria infection.
  • the present disclosure also provides methods. methods. In some embodiments, a method comprises administering a combination described herein to a subject. In some embodiments, a method described herein is a method of treating a malaria infection. In some embodiments, a method described herein is a method of preventing a malaria infection.
  • a subject has or is at risk of developing a malaria infection.
  • a subject is a human.
  • administration induces an anti-malaria immune response in a subject.
  • use of a combination as described herein can be for treatment of a malaria infection.
  • use of a combination as described herein can be for prevention of a malaria injection.
  • FIG.1 presents an overview of the Plasmodium life cycle.
  • FIGS.2A-2B show antigenic fragments of malarial proteins.
  • FIG.2A depicts epitopes observed from MAS1 and MAS2 strings mapped onto MAS3a and MAS4f.
  • FIG.2B shows antigenic fragments of malarial protein LSA-3.
  • FIG.3 presents a schematic representation of exemplary malarial T cell peptide string constructs containing antigens, as described herein.
  • FIG.4 depicts transfection of a combination including MAS3a and MAS4f into cells to generate detectable protein product. Relative protein expression 24 h after co-transfecting 2.5 ⁇ g each drug product into HEK293T cell lines are shown.
  • FIGS.5A-5F depict activation of T-cells, as assessed by secretion of IFN- ⁇ .
  • FIGS.5A-5D show an assessment of IFN- ⁇ secretion using isolated splenocytes (from mice immunized with different T cell peptide string constructs) incubated with construct specific antigen peptide pools (15mers, 11aa overlap across antigen).
  • FIG.5E depicts a comparison of isolated splenocytes (from mice in group 2 and 3, and splenocytes isolated from mice in group 4) response to specific antigen peptide pools.
  • FIG.5F depicts a comparison of isolated splenocytes (from mice in group 2 and splenocytes isolated from mice in group 1) response to specific antigen peptide pools.
  • FIGS.6A-6H depict activation of T-cells, as assessed by secretion of IFN- ⁇ .
  • FIGS.6A-6H show an assessment of IFN- ⁇ secretion using isolated splenocytes (from mice immunized with different T cell peptide string constructs) incubated with construct specific antigen peptide pools (15mers, 11aa overlap across antigen).
  • FIGS.7A-7B depict assessment of activation of T-cells, as assessed by secretion of IFN- ⁇ using isolated splenocytes (from mice immunized with T cell peptide string constructs individually or with T cell strings constructs in combination).
  • FIGS.8A-8B depict assessment of activation of T-cells, as assessed by secretion of IFN- ⁇ using isolated splenocytes (from mice immunized with shorter T cell peptide string constructs or a longer T cell peptide string with the same antigenic content).
  • FIG.9A shows transfection rate as measured by percentage of total HEK293T population that is positive for presence of expressed protein.
  • FIG.9B shows total expression as measured by median fluorescence intensity of the total HEK293T population for both transfected and non-transfected cells.
  • FIGS.10A-10C depict in vitro expression of formulated RNA constructs in HEK293T cells.
  • FIG.10A shows transfection rate as measured by percentage of total HEK293T population that is positive for presence of expressed protein.
  • FIG.10B shows total expression as measured by median fluorescence intensity of the total HEK293T population for both transfected and non-transfected cells.
  • FIGS.11A-11B depict immunogenicity induced in mice by formulated RNA constructs.
  • FIG.11A shows antibodies to Plasmodium falciparum (Pf) CSP full length protein (“PfCSP-FL”).
  • FIG.11B shows antibodies to PfCSP C-terminal domain (“PfCSP-C term”).
  • FIGS.12A-12B depict binding of antibodies generated from mice immunized with formulated RNA constructs to various epitopes.
  • FIG.12A shows a visual summary of the data in form of a heatmap.
  • FIG.12B shows bars that are representative of the area under the curve (AUC) created when plotting dilution steps versus ECL signal.
  • FIGS.13A-13B depict binding specificity of antibodies generated from mice immunized with formulated RNA constructs to CSP protein in Plasmodium falciparum sporozoite lysates.
  • FIG.13A shows binding between antibodies and CSP protein in the sporozoite (spz) lysates, represented as area under the curve (AUC), as assessed by luminescence.
  • FIG.13B shows binding of murine anti-Pfs25 mAb32F81 used as negative control and murine anti-CSP mAb3SP2 used as a positive control.
  • FIGS.14A-14B depict assessment of antibodies generated from mice immunized with formulated RNA constructs for ability to inhibit P. falciparum sporozoite traversal.
  • FIG.14A shows results as the dilution at which the % of traversal is reduced by 50% (mean with SEM).
  • FIG.14B shows the percentage of traversed cells for the negative control (serum from vehicle mice) and the percentage of inhibition of traversal for the positive control (mAb317, an antibody that binds to NANP (SEQ ID NO: 230) repeats of the major repeat region and is known to inhibit traversal), with 002, 003, 005, 012, 014, and 018 indicating different experimental runs.
  • FIGS.15A-15B depict assessment of antibodies generated from mice immunized with formulated RNA constructs for ability to inhibit P. falciparum sporozoite infection of primary human hepatocytes.
  • FIG.15A shows results as percentage of inhibition of infection activity (mean with SEM) in comparison to the average of the vehicle control, which was set as 0% inhibition for three different dilutions of the immune serum.
  • FIG.15B shows the percentage of infected cells from negative control (serum from vehicle mice) and the percentage of inhibition of the positive control (mAb317, an antibody known to inhibit hepatocyte infection).
  • FIG.16 depicts activation of T-cells, as assessed by secretion of IFN- ⁇ .
  • IFN- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (PfCSP_FL_pep), peptides of epitopes predicted to be presented on MHC-I, on MHC-II, or controls (e.g., negative control: gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]), 4 ⁇ g/mL; positive control: concanavalin A, 2 ⁇ g/mL).
  • negative control gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]
  • concanavalin A 2 ⁇ g/mL
  • FIG.17 depicts activation of T-cells, as assessed by secretion of TNF- ⁇ .
  • TNF- ⁇ secretion was assessed using isolated splenocytes (from mice immunized formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (PfCSP_FL_pep), peptides of epitopes predicted to be presented on MHC-I, on MHC-II, or controls (e.g., negative control: gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]), 4 ⁇ g/mL; positive control: concanavalin A, 2 ⁇ g/mL).
  • negative control gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]
  • concanavalin A 2 ⁇ g/mL
  • FIG.18 depicts activation of T-cells, as assessed by secretion of IL-2.
  • IL-2 secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (PfCSP_FL_pep), peptides of epitopes predicted to be presented on MHC-I, on MHC-II, or controls (e.g., negative control: gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]), 4 ⁇ g/mL; positive control: concanavalin A, 2 ⁇ g/mL).
  • negative control gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]
  • concanavalin A 2 ⁇ g/mL
  • FIG.19 depicts activation of T-cells, as assessed by secretion of IL-2 and IFN- ⁇ .
  • IL-2 and IFN- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (PfCSP_FL_pep), peptides of epitopes predicted to be presented on MHC-I, on MHC-II, or controls (e.g., negative control: gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]), 4 ⁇ g/mL; positive control: concanavalin A, 2 ⁇ g/mL).
  • negative control gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]
  • concanavalin A 2 ⁇ g/mL
  • FIG.20 depicts activation of T-cells, as assessed by secretion of TNF- ⁇ and IFN- ⁇ .
  • TNF- ⁇ and IFN- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (PfCSP_FL_pep), peptides of epitopes predicted to be presented on MHC-I, on MHC-II, or controls (e.g., negative control: gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]), 4 ⁇ g/mL; positive control: concanavalin A, 2 ⁇ g/mL).
  • negative control gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]
  • concanavalin A 2 ⁇ g/mL
  • FIG.21 depicts activation of T-cells, as assessed by secretion of TNF- ⁇ and IL-2.
  • TNF- ⁇ and IL-2 secretion was assessed using splenocytes (isolated from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (PfCSP_FL_pep), peptides of epitopes predicted to be presented on MHC-I, on MHC-II, or controls (e.g., negative control: gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]), 4 ⁇ g/mL; positive control: concanavalin A, 2 ⁇ g/mL).
  • splenocytes isolated from mice immunized with formulated RNA constructs
  • PfCSP_FL_pep full length CSP protein
  • FIG.22 depicts activation of T-cells, as assessed by secretion of TNF- ⁇ , IL-2 and IFN- ⁇ .
  • TNF- ⁇ , IL-2 and IFN- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (PfCSP_FL_pep), peptides of epitopes predicted to be presented on MHC-I, on MHC-II, or controls (e.g., negative control: gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]), 4 ⁇ g/mL; positive control: concanavalin A, 2 ⁇ g/mL).
  • negative control gp70-AH1 (SPSYVYHQF [SEQ ID NO: 595]
  • concanavalin A 2 ⁇ g/mL
  • FIGS.23A-23C depict protection of mice immunized with formulated RNA constructs against a challenge with PfCSP-expressing P. berghei sporozoites as well as immunogenicity induced by this immunization.
  • FIG.23A depicts percentage of protected mice up to 11 days after challenge with PfCSP-expressing P. berghei sporozoites, for mice immunized with formulated RNA constructs, vehicle, or positive control.
  • FIG.23B and FIG.23C depict endpoint titers against full length PfCSP two weeks after the boost (day 35, FIG.23B) and one day before the challenge (day 49, FIG.23C) for mice immunized with formulated RNA constructs and mice injected with the vehicle only. Mean ⁇ SEM and individual animal values are shown. [0200] FIGS.24A-24B depict protection of mice immunized with formulated RNA constructs against a challenge with PfCSP-expressing P.
  • FIG.24A depicts percentage of protected mice up to 11 days after challenge with PfCSP-expressing P. berghei sporozoites, for mice immunized with formulated RNA constructs, vehicle (saline), or positive control. Mice that received 100 ⁇ g of the 2A10 monoclonal antibody 24 h before the challenge were used as positive control in Experiment 1 and mice immunized with Mosquirix were used as positive control in Experiment 2 and 3.
  • FIG.24B depicts endpoint titers against full length PfCSP two weeks after the boost (day 35) and one day before the challenge (day 49) for mice immunized with formulated RNA constructs and mice injected with the vehicle only. Mean ⁇ SEM and individual animal values are shown. Mice immunized twice IM with 5 ⁇ g of Mosquirix were used as positive control in Experiments 2 and 3. Mice injected with the vehicle were used as negative controls in all experiments. [0201] FIGS.25A-25J depict assessment of antibodies generated from mice immunized with a formulated RNA construct for ability to recognize native PfCSP on sporozoites and inhibit sporozoite viability and motility. FIG.
  • 25A shows log of anti-sporozoite endpoint titers using fixed PfCSP-expressing P. berghei sporozoites. Symbols represent the mean ⁇ SEM using serum from individual mice.
  • FIG.25B shows inhibition of sporozoite gliding speed. Circles represent the mean ⁇ SEM of duplicates of pooled serum samples from each group.
  • FIG.25C shows estimated length of the circumsporozoite precipitation reaction (CSPR) elicited by serum samples from immunized mice as measured by flow cytometry (Forward Scatter Width (FSC-W)). Symbols represent the mean ⁇ SEM using serum from individual mice.
  • CSPR circumsporozoite precipitation reaction
  • FIG.25D shows cytotoxicity of serum samples from immunized mice against sporozoites in suspension (PBS). Symbols represent the mean ⁇ SEM using serum from individual mice.
  • FIG.25E shows cytotoxicity in 3D (Matrigel). Symbols represent the mean ⁇ SEM using serum from individual mice.
  • FIG.25F depicts 3 experiments and shows log of anti-sporozoite endpoint titers using fixed PfCSP-expressing P. berghei sporozoites. Bars represent the mean ⁇ SEM using serum from individual mice. 2A10, positive antibody control; Mos, Mosquirix® positive control; IFA, Immunofluorescence assay.
  • FIG.25G depicts assessment of antibodies generated from mice immunized with formulated RNA of 5 priority constructs for ability to inhibit sporozoite gliding motility. Each graph represents a different experiment and shows sporozoite gliding speed ( ⁇ m/s). Bars represent the mean ⁇ SEM of duplicates (in Experiment 1) or single replicates (Experiments 2 and 3) of pooled serum samples from each group. 2A10, positive antibody control; Mos, Mosquirix® positive control; Veh, vehicle.
  • FIG.25H depicts assessment of antibodies generated from mice immunized with formulated RNA for ability to bind and crosslink native PfCSP on the sporozoite surface.
  • FIG.25I depicts the cytotoxicity of 17% immune sera measured against PfCSP-expressing P. berghei sporozoites in suspension and is presented as percentage of viable sporozoites.
  • FIG.25J depicts cytotoxicity of 17% immune sera against PfCSP-expressing P.
  • FIGS.25F- 25J each graph represents an independent challenge experiment (designated Experiment 1, Experiment 2 and Experiment 3). Bars represent the mean ⁇ SEM using serum from individual mice. 2A10, positive antibody control; Mosquirix® positive control; Veh, vehicle; PBS, phosphate buffer saline.
  • FIGS.26A- 26B depict in vitro expression of non-formulated RNA constructs encoding different malarial peptide constructs in HEK293T cells.
  • FIG.26A shows transfection rate as measured by percentage of total HEK293T population that is positive for presence of expressed protein.
  • FIG.26B shows total expression as measured by median fluorescence intensity of the total HEK293T population for both transfected and non-transfected cells.
  • FIGS.27A-27B depict in vitro expression of formulated RNA constructs encoding different malarial peptide constructs in HEK293T cells.
  • FIG.27A shows transfection rate as measured by percentage of total HEK293T population that is positive for presence of expressed protein.
  • FIG.27B shows total expression as measured by median fluorescence intensity of the total HEK293T population for both transfected and non-transfected cells.
  • FIGS.28A-28B depict immunogenicity induced in mice by formulated RNA constructs at day 21 after immunization.
  • FIG.28A shows antibodies to an exemplary Plasmodium falciparum (Pf) CSP full length protein (“PfCSP-FL”).
  • FIG.28B shows antibodies to an exemplary PfCSP C-terminal domain (“PfCSP-C term (3D7)”). Each data point is representative of one mouse and the bar denotes mean with SEM. LDL, lower detection limit.
  • FIGS.29A-29B depict immunogenicity induced in mice by formulated RNA constructs at day 35 after immunization.
  • FIG.29A shows antibodies to an exemplary Plasmodium falciparum (Pf) CSP full length protein (“PfCSP-FL”).
  • FIG.29B shows antibodies to an exemplary PfCSP C-terminal domain (“PfCSP-C term (3D7)”). Each data point is representative of one mouse and the bar denotes mean with SEM. LDL, lower detection limit.
  • FIGS.30A-30K depict binding of antibodies generated from mice immunized with formulated RNA constructs to various epitopes.
  • FIG.30A shows a visual summary of the data in FIGS.30B-30K in the form of a heatmap.
  • FIGS.30B-30K show bars that are representative of the area under the curve (AUC) created when plotting dilution steps versus ECL signal.
  • AUC area under the curve
  • FIGS.31A-31B depict binding specificity of antibodies generated from mice immunized with formulated RNA constructs to CSP protein in Plasmodium falciparum sporozoite lysates.
  • FIG.31A shows binding between antibodies in the sera of immunized mice and CSP protein in the sporozoite (spz) lysates represented as area under the curve (AUC) created when plotting dilution steps versus luminescence signal.
  • FIG.31B shows binding of murine anti-CSP mAb3SP2 used as a positive control.
  • FIGS.32A-32C depict activation of T-cells, as assessed by secretion of IFN- ⁇ .
  • IFN- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering a full length CSP protein (FIG.32A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG. 32B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.32C)). Samples were measured in triplicate and negative control was measured in duplicate; each data point represents a single mouse and bars represent the group mean spot-forming units (SFU) ⁇ SD per 5x10 5 splenocytes. Each data point in the medium and ConA controls represents the mean of triplicates of a pool of splenocytes from all mice.
  • SFU group mean spot-forming units
  • FIGS.33A-33C depict activation of T-cells, as assessed by secretion of IL-2.
  • IL-2 secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering a full length CSP protein (FIG.33A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG. 33B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.33C)).
  • Samples were measured in triplicate and negative control was measured in duplicate; each data point represents a single mouse and bars represent the group mean spot-forming units (SFU) ⁇ SD per 5x10 5 splenocytes.
  • SFU spot-forming units
  • FIGS.34A-34C depict activation of T-cells, as assessed by secretion of TNF- ⁇ .
  • TNF- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering a full length CSP protein (FIG.34A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG. 34B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.34C)).
  • FIGS.35A-35C depict activation of T-cells, as assessed by secretion of both IFN- ⁇ and IL-2.
  • IFN- ⁇ +IL- 2 secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering a full length CSP protein (FIG.35A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.35B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.35C)). Samples were measured in triplicate and negative control was measured in duplicate; each data point represents a single mouse and bars represent the group mean spot-forming units (SFU) ⁇ SD per 5x10 5 splenocytes.
  • SFU spot-forming units
  • FIGS.36A-36C depict activation of T-cells, as assessed by secretion of both IFN- ⁇ and TNF- ⁇ .
  • IFN- ⁇ + TNF- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering a full length CSP protein (FIG.36A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.36B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.36C)).
  • FIGS.37A-37C depict activation of T-cells, as assessed by secretion of both IL-2 and TNF- ⁇ .
  • IL-2+ TNF- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering a full length CSP protein (FIG.37A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.37B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.37C)). Samples were measured in triplicate and negative control was measured in duplicate; each data point represents a single mouse and bars represent the group mean spot-forming units (SFU) ⁇ SD per 5x10 5 splenocytes.
  • SFU spot-forming units
  • FIGS.38A-38C depict activation of T-cells, as assessed by secretion of IFN- ⁇ , IL-2 and TNF- ⁇ .
  • IFN- ⁇ +IL-2+TNF- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering a full length CSP protein (FIG.38A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.38B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.38C)).
  • FIGS.39A-39D depict activation of CD4 T cells only, as assessed by secretion of IFN- ⁇ . IFN- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs).
  • FIGS.40A-40D depict activation of CD4 T cells only, as assessed by secretion of IL-2.
  • IL-2 secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering a full length CSP protein (FIG.40A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG. 40B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.40C); medium control (FIG.40D)).
  • negative control Trp1, 2 ⁇ g/mL
  • FIG.40C positive control
  • FOG.40D medium control
  • FIGS.41A-41D depict activation of CD4 T cells only, as assessed by secretion of TNF- ⁇ . TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs).
  • FIGS.42A-42D depict activation of CD4 T cells only, as assessed by secretion of both IFN- ⁇ and IL-2.
  • IFN- ⁇ +IL-2 secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering a full length CSP protein (FIG.42A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.42B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.42C); medium control (FIG.42D)).
  • FIGS.43A-43D depict activation of CD4 T cells only, as assessed by secretion of both IFN- ⁇ and TNF- ⁇ . IFN- ⁇ +TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs).
  • FIGS.44A-44D depict activation of CD4 T cells only, as assessed by secretion of both IL-2 and TNF- ⁇ .
  • IL-2+TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering a full length CSP protein (FIG.44A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.44B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.44C); medium control (FIG.44D)).
  • FIGS.45A-45D depict activation of CD4 T cells only, as assessed by secretion of IFN- ⁇ , IL-2 and TNF- ⁇ . IFN- ⁇ +IL-2+TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs).
  • FIGS.46A-46D depict activation of CD8 T cells only, as assessed by secretion of IFN- ⁇ .
  • IFN- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering a full length CSP protein (FIG.46A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.46B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.46C); medium control (FIG.46D)).
  • FIGS.47A-47D depict activation of CD8 T cells only, as assessed by secretion of IL-2.
  • IL-2 secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering a full length CSP protein (FIG.47A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.
  • FIGS.48A-48D depict activation of CD8 T cells only, as assessed by secretion of TNF- ⁇ . TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs).
  • FIGS.49A-49D depict activation of CD8 T cells only, as assessed by secretion of both IFN- ⁇ and IL-2.
  • IFN- ⁇ +IL-2 secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering a full length CSP protein (FIG.49A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.49B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.49C); medium control (FIG.49D)).
  • FIGS.50A-50D depict activation of CD8 T cells only, as assessed by secretion of both IFN- ⁇ and TNF- ⁇ . IFN- ⁇ +TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs).
  • FIGS.51A-51D depict activation of CD8 T cells only, as assessed by secretion of both IL-2 and TNF- ⁇ .
  • IL-2+TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering a full length CSP protein (FIG.51A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.51B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.51C); medium control (FIG.51D)).
  • negative control Trp1, 2 ⁇ g/mL
  • FIG.51C positive control
  • FOG.51D medium control
  • FIGS.52A-52D depict activation of CD8 T cells only, as assessed by secretion of IFN- ⁇ , IL-2 and TNF- ⁇ . IFN- ⁇ +IL-2+TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with each of the formulated RNA constructs).
  • FIGS.53A-53C depict in vitro expression of ERMA 23-7 RNA construct in HEK293T cells.
  • FIG.53A shows transfection rate as measured by percentage of total HEK293T population that is positive for presence of expressed protein.
  • FIG.53B shows total expression as measured by median fluorescence of the total HEK293T population for both transfected and non-transfected cells.
  • FIG.53C shows percentage of viable cells that are positive for presence of expressed protein, with non-transfected cells serving as a control.
  • FIGS.54A-54B depict titers of antibodies elicited against PfCSP after immunization of mice with a pharmaceutical composition comprising ERMA 23-7.
  • FIG.54A shows endpoint titers against full-length PfCSP on day 21, pre-boost.
  • FIG.54B shows endpoint titers against full-length PfCSP on day 35 after boost.
  • FIGS.55A-55B depict epitope specificity of antibodies elicited upon immunization of mice with a pharmaceutical composition comprising ERMA 23-7.
  • FIG.55A shows a diagram depicting localization of peptides using in multiplex assay in central region of PfCSP sequence.
  • FIG.55B shows binding to epitopes by calculating AUC of 8-point dilution of immune serum samples.
  • FIGS.56A-56C depict pro-inflammatory response from T cells after immunization of mice with a pharmaceutical composition comprising ERMA 23-7.
  • FIG.56A shows production of IFN ⁇ in mouse splenocytes after immunization with a pharmaceutical composition comprising ERMA 23-7 and stimulation with PfCSP peptides.
  • FIG. 56B shows production of IFN ⁇ in combination with IL-2 in mouse splenocytes after immunization with a pharmaceutical composition comprising ERMA 23-7 and stimulation with PfCSP peptides.
  • FIG.56C shows production of IFN ⁇ in combination with IL-2 and TNF ⁇ in mouse splenocytes after immunization with a pharmaceutical composition comprising ERMA 23-7 and stimulation with PfCSP peptides.
  • FIGS.57A-57B depict titers of antibodies elicited against PfCSP after immunization of mice with a pharmaceutical composition comprising ERMA 23-7 or with Composition 1, 2, and 3 comprising ERMA 23-7, MAS3a and MAS4f.
  • FIG.57A shows endpoint titers (reciprocal serum titer) against full-length PfCSP pre-boost on Day 21.
  • FIG.57B shows endpoint titers (reciprocal serum titer) against full-length PfCSP after boost on Day 35.
  • FIG.58 depicts epitope specificity of antibodies elicited upon immunization of mice with a pharmaceutical composition comprising ERMA 23-7 or with Compositions 1, 2 and 3 comprising ERMA 23-7, MAS3a and MAS4f.
  • FIG.59 depicts T-cell induction following immunization with Composition 3 comprising 1 ⁇ g ERMA 23- 7, 2 ⁇ g MAS3a and 2 ⁇ g MAS4f.
  • FIGS.60A-60B depict T-cell induction following immunization.
  • FIG.60A shows the results for a combination of two T-cell string constructs by comparing the effects of a pharmaceutical composition comprising 2 ⁇ g MAS3a and 2 ⁇ g MAS4f (black dots) and Composition 3 comprising 1 ⁇ g ERMA 23-7, 2 ⁇ g MAS3a and 2 ⁇ g MAS4f (red dots).
  • FIG.60B shows the results for a CSP construct by comparing the effects of a pharmaceutical composition comprising 1 ⁇ g ERMA 23-7 (black dots) and Composition 3 comprising 1 ⁇ g ERMA 23-7, 2 ⁇ g MAS3a and 2 ⁇ g MAS4f (red dots).
  • FIGS.61A-61K depict an assessment of antibodies generated from mice immunized with formulated RNA constructs for their ability to inhibit P. falciparum sporozoite infection of primary human hepatocytes. Percentage of inhibition of infection activity (mean with SEM) in comparison to a control (medium only) is shown for a 1:40 dilution (FIGS.61A and 61E), a 1:160 dilution (FIGS.61B and 61F), a 1:640 dilution (FIGS.61C and 61G), and a 1:2560 dilution (FIGS.61D and 61H).
  • FIGS. 62A-62F depict an assessment of binding and disassociation of antibodies generated from mice immunized with formulated RNA constructs and exposed to full length PfCSP, a peptide with a junction region and minor repeats (Junction + Minor repeats), or a peptide with major repeats (Major repeats).
  • FIGS.63A-63D depict T-cell induction following immunization with 1 ⁇ g of Mas3a or a codon- optimized version (Mas3a-2, Mas3a-3) and 1 ⁇ g Mas4f or a codon-optimized version (Mas4f-2, Mas4f-3).
  • Antigen specific T-cell induction 7 days after immunization is shown for the combination of 1 ⁇ g Mas3a and 1 ⁇ g Mas4f (FIG. 63A), and the combination of 1 ⁇ g Mas3a-3 and 1 ⁇ g Mas4f-3 (FIG.63B).
  • Antigen specific T-cell induction 35 days after immunization is shown for the combination of 1 ⁇ g Mas3a and 1 ⁇ g Mas4f (FIG.63C), and the combination of 1 ⁇ g Mas3a-3 and 1 ⁇ g Mas4f-3 (FIG.63D).
  • FIGS.64A-64D depict antigen-specific IFN ⁇ and IL-2 T-cell responses following immunization with 1 ⁇ g of Mas3a or a codon-optimized version (Mas3a-2, Mas3a-3) and 1 ⁇ g Mas4f or a codon-optimized version (Mas4f- 2, Mas4f-3).
  • Antigen-specific IFN ⁇ responses after immunization for the combination of 1 ⁇ g Mas3a and 1 ⁇ g Mas4f is compared to the combination of 1 ⁇ g Mas3a-3 and 1 ⁇ g Mas4f-3 (FIG.64A) and Mas3a-2 and 1 ⁇ g Mas4f-2 (FIG. 64C).
  • FIGS.65A-65H depict assessment of antibodies generated from mice immunized with formulated RNA constructs for ability to inhibit P. falciparum sporozoite traversal in HC-04 hepatoma cells.
  • Percentage of inhibition of traversal activity (mean with SEM) in comparison to a medium control, which was set as 0% inhibition, is shown for a 1:20 dilution (FIG.65A), a 1:40 dilution (FIG.65B), a 1:80 dilution (FIG.65C), a 1:160 dilution (FIG. 65D), a 1:320 dilution (FIG.65E), and a 1:640 dilution (FIG.65F).
  • Inhibition of antibodies in sera of immunized mice in traversal assays is represented as area under the curve (AUC) created when plotting dilution steps versus % inhibition of traversal (FIG.65G).
  • FIGS.66A-66G depict assessment of antibodies generated from mice immunized with formulated RNA constructs for ability to inhibit P. falciparum sporozoite traversal in HC-04 hepatoma cells.
  • Percentage of inhibition of traversal activity (mean with SEM) in comparison to a medium control, which was set as 0% inhibition, is shown for a 1:20 dilution (FIG.66A), a 1:40 dilution (FIG.66B), a 1:80 dilution (FIG.66C), a 1:160 dilution (FIG.66D), a 1:320 dilution (FIG.66E), and a 1:640 dilution (FIG.66F).
  • Inhibition of antibodies in sera of immunized mice in traversal assays is represented as area under the curve (AUC) created when plotting dilution steps versus % inhibition of traversal (FIG.66G).
  • FIGS.67A-67B depict in vitro expression of non-formulated RNA constructs 91, 100 and 104 encoding different Plasmodium polypeptides in HEK293T cells.
  • FIG.67A shows transfection rate as measured by percentage of total HEK293T population that is positive for presence of expressed protein.
  • FIG.67B shows total expression as measured by median fluorescence of the total HEK293T population for both transfected and non-transfected cells.
  • Permeabilized cells show total protein expressed (black bar, intracellular staining) and non-permeabilized cells show only surface expressed protein (grey bar, surface staining). Protein was detected using anti-PfCSP L9 antibody.
  • FIGS.68A-68B depict in vitro expression of non-formulated RNA constructs 87 and 88 encoding different Plasmodium polypeptides in HEK293T cells.
  • FIG.68A shows transfection rate as measured by percentage of total HEK293T population that is positive for presence of expressed protein.
  • FIG.68B shows total expression as measured by median fluorescence of the total HEK293T population for both transfected and non-transfected cells.
  • Permeabilized cells show total protein expressed (black bar, intracellular staining) and non-permeabilized cells show only surface expressed protein (grey bar, surface staining).
  • FIGS.69A-69B depict in vitro expression of formulated RNA constructs 87, 88, 91, 100 and 104 encoding different Plasmodium polypeptides in HEK293T cells.
  • FIG.69A shows transfection rate as measured by percentage of total HEK293T population that is positive for presence of expressed protein.
  • FIG.69B shows total expression as measured by median fluorescence of the total HEK293T population for both transfected and non- transfected cells.
  • FIGS.70A-70B depict immunogenicity induced in mice by formulated RNA constructs 87, 88, 91, 100 and 104.
  • FIG.70A shows antibodies to Plasmodium falciparum (Pf) CSP full length protein (“PfCSP-FL”).
  • FIG.70B shows antibodies to PfCSP C-terminal domain (“PfCSP-C term (3D7)”).
  • FIG.71 depicts binding of antibodies generated from mice immunized with formulated RNA constructs 87, 88, 91, 104, and 100 during challenge studies to various epitopes in a heatmap format.
  • FIGS.72A-72J depict binding of antibodies generated from mice immunized with formulated RNA constructs to various epitopes.
  • FIGS.72A-72J each show bars that are representative of the area under the curve (AUC) created when plotting dilution steps versus ECL signal.
  • FIGS.73A-73C depict activation of T-cells, as assessed by secretion of IFN- ⁇ .
  • IFN- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (FIG.73A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.73B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.73C)). Samples were measured in triplicate and negative control was measured in duplicate; each data point represents a single mouse and bars represent the group mean spot-forming units (SFU) ⁇ SD per 5x10 5 splenocytes. Each data point in the medium and ConA controls represents the mean of triplicates of a pool of splenocytes from all mice.
  • SFU group mean spot-forming units
  • FIGS.74A-74C depicts activation of T-cells, as assessed by secretion of IL-2.
  • IL-2 secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (FIG.74A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.74B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.74C)).
  • FIGS.75A-75C depicts activation of T-cells, as assessed by secretion of TNF- ⁇ .
  • TNF- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (FIG.75A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.75B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.75C)). Samples were measured in triplicate and negative control was measured in duplicate; each data point represents a single mouse and bars represent the group mean spot-forming units (SFU) ⁇ SD per 5x10 5 splenocytes. Each data point in the medium and ConA controls represents the mean of triplicates of a pool of splenocytes from all mice.
  • SFU group mean spot-forming units
  • FIGS.76A-76C depicts activation of T-cells, as assessed by secretion of both IFN- ⁇ and IL-2.
  • IFN- ⁇ +IL-2 secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (FIG.76A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.76B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.76C)).
  • FIGS.77A-77C depicts activation of T-cells, as assessed by secretion of both IFN- ⁇ and TNF- ⁇ .
  • IFN- ⁇ + TNF- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (FIG.77A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.77B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.77C)). Samples were measured in triplicate and negative control was measured in duplicate; each data point represents a single mouse and bars represent the group mean spot-forming units (SFU) ⁇ SD per 5x10 5 splenocytes.
  • SFU spot-forming units
  • FIGS.78A-78C depicts activation of T-cells, as assessed by secretion of both IL-2 and TNF- ⁇ .
  • IL-2+ TNF- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (FIG.78A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.78B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.78C)).
  • FIGS.79A-79C depicts activation of T-cells, as assessed by secretion of IFN- ⁇ , IL-2 and TNF- ⁇ .
  • IFN- ⁇ +IL-2+TNF- ⁇ secretion was assessed using isolated splenocytes (from mice immunized with formulated RNA constructs) treated with overlapping peptide pools covering the full length CSP protein (FIG.79A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.79B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.79C)). Samples were measured in triplicate and negative control was measured in duplicate; each data point represents a single mouse and bars represent the group mean spot-forming units (SFU) ⁇ SD per 5x10 5 splenocytes.
  • SFU spot-forming units
  • FIGS.80A-80D depicts activation of CD4 T cells only, as assessed by secretion of IFN- ⁇ . IFN- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs).
  • FIGS.81A-81D depicts activation of CD4 T cells only, as assessed by secretion of IL-2.
  • IL-2 secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering the full length CSP protein (FIG.81A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.81B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.81C); medium control (FIG.81D)).
  • negative control Trp1, 2 ⁇ g/mL
  • FIG.81C positive control
  • concanavalin A 2 ⁇ g/mL
  • FIG.81D medium control
  • FIGS.82A-82D depicts activation of CD4 T cells only, as assessed by secretion of TNF- ⁇ . TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs).
  • FIGS.83A-83D depicts activation of CD4 T cells only, as assessed by secretion of both IFN- ⁇ and IL- 2.
  • IFN- ⁇ +IL-2 secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering the full length CSP protein (FIG.83A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.83B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.83C); medium control (FIG.83D)).
  • negative control Trp1, 2 ⁇ g/mL
  • FAC concanavalin A
  • FIG.83D medium control
  • FIGS.84A-84D depicts activation of CD4 T cells only, as assessed by secretion of both IFN- ⁇ and TNF- ⁇ . IFN- ⁇ +TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs).
  • FIGS.85A-85D depicts activation of CD4 T cells only, as assessed by secretion of both IL-2 and TNF- ⁇ .
  • IL-2+TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering the full length CSP protein (FIG.85A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.85B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.85C); medium control (FIG.85D)).
  • negative control Trp1, 2 ⁇ g/mL
  • FIG.85C positive control
  • FOG.85D medium control
  • FIGS.86A-86D depicts activation of CD4 T cells only, as assessed by secretion of IFN- ⁇ , IL-2 and TNF- ⁇ . IFN- ⁇ +IL-2+TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD4+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs).
  • FIGS.87A-87D depicts activation of CD8 T cells only, as assessed by secretion of IFN- ⁇ .
  • IFN- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering the full length CSP protein (FIG.87A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.87B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.87C); medium control (FIG.87D)).
  • negative control Trp1, 2 ⁇ g/mL
  • FAC concanavalin A
  • FIG.87D medium control
  • FIGS.88A-88D depicts activation of CD8 T cells only, as assessed by secretion of IL-2. IL-2 secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs).
  • FIGS.89A-89D depicts activation of CD8 T cells only, as assessed by secretion of TNF- ⁇ .
  • TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering the full length CSP protein (FIG.89A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.89B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.89C); medium control (FIG.89D)).
  • negative control Trp1, 2 ⁇ g/mL
  • FAC concanavalin A
  • FIG.89D medium control
  • FIGS.90A-90D depicts activation of CD8 T cells only, as assessed by secretion of both IFN- ⁇ and IL- 2.
  • IFN- ⁇ +IL-2 secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs).
  • FIGS.91A-91D depicts activation of CD8 T cells only, as assessed by secretion of both IFN- ⁇ and TNF- ⁇ .
  • IFN- ⁇ +TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering the full length CSP protein (FIG.91A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.91B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.91C); medium control (FIG.91D)).
  • FIGS.92A-92D depicts activation of CD8 T cells only, as assessed by secretion of both IL-2 and TNF- ⁇ .
  • IL-2+TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs).
  • FIGS.93A-93D depicts activation of CD8 T cells only, as assessed by secretion of IFN- ⁇ , IL-2 and TNF- ⁇ .
  • IFN- ⁇ +IL-2+TNF- ⁇ secretion was assessed by a fluorospot assay after using MACS separation to isolate CD8+ T cells (from pools of splenocytes from mice immunized with formulated RNA constructs). Cells were then incubated with overlapping peptide pools covering the full length CSP protein (FIG.93A) or controls (e.g., negative control: Trp1, 2 ⁇ g/mL (FIG.93B); positive control: concanavalin A, 2 ⁇ g/mL (FIG.93C); medium control (FIG.93D)).
  • FIG.94 shows binding between antibodies in the sera of immunized mice and CSP protein in the sporozoite (spz) lysates represented as area under the curve (AUC) created when plotting dilution steps versus luminescence signal.
  • AUC area under the curve
  • each stereocenter the R and S configurations of each stereocenter are contemplated as part of the disclosure. Therefore, single stereochemical isomers, as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of provided compounds are within the scope of the disclosure.
  • provided compounds show one or more stereoisomers of a compound, and unless otherwise indicated, represents each stereoisomer alone and/or as a mixture. Unless otherwise stated, all tautomeric forms of provided compounds are within the scope of the disclosure. [0273] Unless otherwise indicated, structures depicted herein are meant to include compounds that differ only in the presence of one or more isotopically enriched atoms.
  • the term “about” may encompass a range of values that within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referred value.
  • agent may refer to a physical entity.
  • an agent may be characterized by a particular feature and/or effect.
  • therapeutic agent refers to a physical entity has a therapeutic effect and/or elicits a desired biological and/or pharmacological effect.
  • an agent may be a compound, molecule, or entity of any chemical class including, for example, a small molecule, polypeptide, nucleic acid, saccharide, lipid, metal, or a combination or complex thereof.
  • Amino acid In its broadest sense, as used herein, the term “amino acid” refers to a compound and/or substance that can be, is, or has been incorporated into a polypeptide chain, e.g., through formation of one or more polypeptide bonds.
  • an amino acid has the general structure H2N–C(H)(R)–COOH.
  • an amino acid is a naturally-occurring amino acid.
  • an amino acid is a non- natural amino acid; in some embodiments, an amino acid is a D-amino acid; in some embodiments, an amino acid is an L-amino acid.
  • Standard amino acid refers to any of the twenty standard L-amino acids commonly found in naturally occurring polypeptides.
  • Nonstandard amino acid refers to any amino acid, other than the standard amino acids, regardless of whether it is prepared synthetically or obtained from a natural source.
  • an amino acid, including a carboxy- and/or amino-terminal amino acid in a polypeptide can contain a structural modification as compared with the general structure above.
  • an amino acid may be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and/or substitution (e.g., of the amino group, the carboxylic acid group, one or more protons, and/or the hydroxyl group) as compared with the general structure.
  • such modification may, for example, alter the circulating half-life of a polypeptide containing the modified amino acid as compared with one containing an otherwise identical unmodified amino acid.
  • such modification does not significantly alter a relevant activity of a polypeptide containing the modified amino acid, as compared with one containing an otherwise identical unmodified amino acid.
  • amino acid may be used to refer to a free amino acid; in some embodiments it may be used to refer to an amino acid residue of a polypeptide.
  • Antigen refers to an agent that elicits an immune response; and/or (ii) an agent that binds to a T cell receptor (e.g., when presented by an MHC molecule) or to an antibody.
  • malaria antigen and “Plasmodium antigen” are understood to refer to an antigen from a Plasmodium species, where the Plasmodium species can cause malaria in a subject.
  • Anti-malaria immune response refers to an immune response produced through pre-exposure to one or more Plasmodium antigens, e.g., through administration (e.g., vaccination) of the constructs as described herein directed to Plasmodium.
  • Associated Two events or entities are “associated” with one another, as that term is used herein, if the presence, level, degree, type and/or form of one is correlated with that of the other.
  • a particular entity e.g., polypeptide, genetic signature, metabolite, microbe, etc.
  • a particular entity e.g., polypeptide, genetic signature, metabolite, microbe, etc.
  • a particular disease, disorder, or condition if its presence, level and/or form correlates with incidence of, susceptibility to, severity of, stage of, etc. the disease, disorder, or condition (e.g., across a relevant population).
  • two or more entities are physically “associated” with one another if they interact, directly or indirectly, so that they are and/or remain in physical proximity with one another.
  • C-terminal domain refers to a region of a CSP polypeptide that corresponds to amino acids 273-397 of wild-type CSP sequence of Plasmodium falciparum (isolate 3D7) (SEQ ID NO: 1).
  • C-terminal region refers to a region of a CSP polypeptide that corresponds to amino acids 273-375 of wild-type CSP sequence (SEQ ID NO: 1). In some embodiments, a serine follows immediately after the C-terminal region. In some embodiments, a serine and a valine follow immediately after the C-terminal region. [0282] C-terminal region variant: The term “C-terminal region variant”, as used herein, refers to a C- terminal region that comprises one or more mutations as compared to amino acids 273-375 of wild-type CSP sequence (SEQ ID NO: 1). In some embodiments, one or more mutations are one or more substitution mutations.
  • one or more mutations comprise an indel.
  • Central domain The term “central domain”, as used herein, refers to a region of a CSP polypeptide that corresponds to amino acids 105-272 of wild-type CSP sequence (SEQ ID NO: 1).
  • Characteristic portion As used herein, the term “characteristic portion”, in the broadest sense, refers to a portion of a polypeptide or region thereof whose presence (or absence) correlates with presence (or absence) of a particular feature, attribute, or activity of the polypeptide or region thereof.
  • a characteristic portion of a polypeptide or region thereof is a portion that is found in the polypeptide or region thereof and in related polypeptide or region thereof that share the particular feature, attribute or activity, but not in those that do not share the particular feature, attribute or activity. In certain embodiments, a characteristic portion shares at least one functional characteristic with the intact polypeptide or region thereof.
  • a “characteristic portion” of a polypeptide or region thereof is one that contains a continuous stretch of amino acids, or a collection of continuous stretches of amino acids, that together are characteristic of the polypeptide or region thereof. In some embodiments, each such continuous stretch generally contains at least 2, 5, 10, 15, 20, 50, or more amino acids.
  • a characteristic portion of a polypeptide or region thereof is one that, in addition to the sequence and/or structural identity specified above, shares at least one functional characteristic with the relevant intact polypeptide or region thereof.
  • a characteristic portion may be biologically active.
  • a fragment as described herein can be a portion. Accordingly, in some embodiments, a characteristic fragment can be a “characteristic portion.”
  • Combination therapy refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents (e.g., two or more antibody agents)).
  • the two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all “doses” of a first regimen are administered prior to administration of any doses of a second regimen); in some embodiments, such agents are administered in overlapping dosing regimens.
  • administration of combination therapy may involve administration of one or more agent(s) or modality(ies) to a subject receiving the other agent(s) or modality(ies) in the combination.
  • combination therapy does not require that individual agents be administered together in a single composition (or even necessarily at the same time), although in some embodiments, two or more agents, or active moieties thereof, may be administered together in a combination composition.
  • Comparable refers to two or more agents, entities, situations, sets of conditions, etc., that may not be identical to one another but that are sufficiently similar to permit comparison there between so that one skilled in the art will appreciate that conclusions may reasonably be drawn based on differences or similarities observed.
  • comparable sets of conditions, circumstances, individuals, or populations are characterized by a plurality of substantially identical features and one or a small number of varied features.
  • corresponding to refers to a relationship between two or more entities.
  • corresponding to may be used to designate the position/identity of a structural element in a compound or composition relative to another compound or composition (e.g., to an appropriate reference compound or composition).
  • a monomeric residue in a polymer may be identified as “corresponding to” a residue in an appropriate reference polymer.
  • residues in a polypeptide are often designated using a canonical numbering system based on a reference related polypeptide, so that an amino acid “corresponding to” a residue at position 190, for example, need not actually be the 190th amino acid in a particular amino acid chain but rather corresponds to the residue found at 190 in the reference polypeptide; those of ordinary skill in the art readily appreciate how to identify “corresponding” amino acids.
  • sequence alignment strategies including software programs such as, for example, BLAST, CS-BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH/GLSEARCH, Genoogle, HMMER, HHpred/HHsearch, IDF, Infernal, KLAST, USEARCH, parasail, PSI-BLAST, PSI-Search, ScalaBLAST, Sequilab, SAM, SSEARCH, SWAPHI, SWAPHI-LS, SWIMM, or SWIPE that can be utilized, for example, to identify “corresponding” residues in polypeptide and/or nucleic acids in accordance with the present disclosure.
  • software programs such as, for example, BLAST, CS-BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH/GLSEARCH, Genoogle, HMMER, HHpred/HHsearch, IDF, Infernal, KLAST, USEARCH, parasail, PSI-BLAST, PSI-Search, ScalaBL
  • corresponding to may be used to describe an event or entity that shares a relevant similarity with another event or entity (e.g., an appropriate reference event or entity).
  • a gene or protein in one organism may be described as “corresponding to” a gene or protein from another organism in order to indicate, in some embodiments, that it plays an analogous role or performs an analogous function and/or that it shows a particular degree of sequence identity or homology, or shares a particular characteristic sequence element.
  • Dosing regimen may be used to refer to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time. In some embodiments, a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses.
  • Encode As used herein, the term “encode” or “encoding” refers to sequence information of a first molecule that guides production of a second molecule having a defined sequence of nucleotides (e.g., a polyribonucleotide) or a defined sequence of amino acids.
  • a DNA molecule can encode an RNA molecule (e.g., by a transcription process that includes a DNA-dependent RNA polymerase enzyme).
  • An RNA molecule can encode a polypeptide (e.g., by a translation process).
  • a gene, a cDNA, or an RNA molecule encodes a polypeptide if transcription and translation of RNA corresponding to that gene produces the polypeptide in a cell or other biological system.
  • a coding region of a polyribonucleotide encoding a target antigen refers to a coding strand, the nucleotide sequence of which is identical to the polyribonucleotide sequence of such a target antigen.
  • a coding region of a polyribonucleotide encoding a target antigen refers to a non-coding strand of such a target antigen, which may be used as a template for transcription of a gene or cDNA.
  • expression As used herein, the term “expression” of a nucleic acid sequence refers to the generation of a gene product from the nucleic acid sequence.
  • a gene product can be a transcript, e.g., a polyribonucleotide as provided herein.
  • a gene product can be a polypeptide.
  • expression of a nucleic acid sequence involves one or more of the following: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of an RNA transcript (e.g., by splicing, editing, etc.); (3) translation of an RNA into a polypeptide or protein; and/or (4) post-translational modification of a polypeptide or protein.
  • helper antigen refers to an antigen that is included in a polypeptide comprising one or more CSP polypeptide regions or portion thereof, where the antigen is not derived from a CSP polypeptide.
  • Heterologous As used herein, the term “heterologous”, with respect to secretory signal or transmembrane region, refers to a secretory signal or transmembrane region from a virus or an organism other than Plasmodium.
  • Homology As used herein, the term “homology” or “homolog” refers to the overall relatedness between polynucleotide molecules (e.g., DNA molecules and/or RNA molecules) and/or between polypeptide molecules.
  • polynucleotide molecules e.g., DNA molecules and/or RNA molecules
  • polypeptide molecules are considered to be “homologous” to one another if their sequences are at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical.
  • polynucleotide molecules e.g., DNA molecules and/or RNA molecules
  • polypeptide molecules are considered to be “homologous” to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% similar (e.g., containing residues with related chemical properties at corresponding positions).
  • certain amino acids are typically classified as similar to one another as “hydrophobic” or “hydrophilic” amino acids, and/or as having “polar” or “non-polar” side chains.
  • Identity refers to the overall relatedness between polynucleotide molecules (e.g., DNA molecules and/or RNA molecules) and/or between polypeptide molecules.
  • polynucleotide molecules e.g., DNA molecules and/or RNA molecules
  • polypeptide molecules are considered to be “substantially identical” to one another if their sequences are at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical.
  • Calculation of the percent identity of two nucleic acid or polypeptide sequences can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second sequence for optimal alignment and non-identical sequences can be disregarded for comparison purposes).
  • the length of a sequence 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 a reference sequence.
  • the nucleotides at corresponding positions are then compared.
  • the percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences.
  • the comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller, 1989, which has been incorporated into the ALIGN program (version 2.0).
  • nucleic acid sequence comparisons made with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.
  • the percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix.
  • Increased, Induced, or Reduced indicate values that are relative to a comparable reference measurement.
  • an assessed value achieved with a provided composition e.g., a pharmaceutical composition
  • an assessed value achieved in a subject may be “increased” relative to that obtained in the same subject under different conditions (e.g., prior to or after an event; or presence or absence of an event such as administration of a composition (e.g., a pharmaceutical composition) as described herein, or in a different, comparable subject (e.g., in a comparable subject that differs from the subject of interest in prior exposure to a condition, e.g., absence of administration of a composition (e.g., a pharmaceutical composition) as described herein.).
  • comparative terms refer to statistically relevant differences (e.g., that are of a prevalence and/or magnitude sufficient to achieve statistical relevance).
  • the term “reduced” or equivalent terms refers to a reduction in the level of an assessed value by at least 5%, at least 10%, at least 20%, at least 50%, at least 75% or higher, as compared to a comparable reference. In some embodiments, the term “reduced” or equivalent terms refers to a complete or essentially complete inhibition, i.e., a reduction to zero or essentially to zero.
  • the term “increased” or “induced” refers to an increase in the level of an assessed value by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 80%, at least 100%, at least 200%, at least 500%, or higher, as compared to a comparable reference.
  • “in order” refers to the order of features from 5' to 3' along the polynucleotide or polyribonucleotide.
  • Isolated means altered or removed from the natural state.
  • junction refers to a region of a CSP polypeptide that corresponds to amino acids 98-104 of wild-type CSP sequence (SEQ ID NO: 1).
  • junction region refers to a region of a CSP polypeptide that corresponds to amino acids 93-104 of wild-type CSP sequence (SEQ ID NO: 1). Typically, a junction region includes an R1 region (amino acids 93-97) and a junction (SEQ ID NO: 277) at positions 98-104.
  • Junction region variant refers to a junction region that comprises one or more mutations as compared to amino acids 93-104 of wild-type CSP sequence (SEQ ID NO: 1). In some embodiments, one or more mutations are one or more substitution mutations.
  • one or more mutations comprise an indel.
  • Linker As used herein, the term “linker” refers to a portion of a polypeptide that connects different regions, portions, or antigens to one another.
  • Lipid As used herein, the terms “lipid” and “lipid-like material” are broadly defined as molecules which comprise one or more hydrophobic moieties or groups and optionally also one or more hydrophilic moieties or groups. Molecules comprising hydrophobic moieties and hydrophilic moieties are also typically denoted as amphiphiles.
  • Major repeat region refers to a region of a CSP polypeptide that corresponds to amino acids 129-272 of wild-type CSP sequence (SEQ ID NO: 1) and contains 35 repeats of the amino acid sequence NANP (SEQ ID NO: 230).
  • the 35 repeats of the amino acid sequence NANP (SEQ ID NO: 230) are separated into two contiguous stretches, the first stretch containing 17 repeats of the amino acid sequence NANP (SEQ ID NO: 230) and second stretch containing 18 repeats of the amino acid sequence NANP (SEQ ID NO: 230) which flank an amino acid sequence of NVDP (SEQ ID NO: 229).
  • a portion of the major repeat region contains at least the amino acid sequence NPNA (SEQ ID NO: 228). Preferably a portion of the major repeat region contains at least the amino acid sequences NANPNA (SEQ ID NO: 232) and NPNANP (SEQ ID NO: 231).
  • “repeat” in reference to sequence A refers to sequence A being present once, and “one or more repeats” of sequence A refers to sequence A being present one or more times.
  • Merozoite stage specific Plasmodium antigen refers to an antigen that is expressed during the merozoite stage of the Plasmodium life cycle.
  • Minor repeat region refers to a region of a CSP polypeptide that corresponds to amino acids 105-128 of wild-type CSP sequence (SEQ ID NO: 1) and contains 3 repeats of the amino acid sequence NANPNVDP (SEQ ID NO: 223).
  • a minor repeat region does not contain the amino acid sequence NPNA (SEQ ID NO: 228), and does not contain the amino acid sequence NANPNA (SEQ ID NO: 232) or NPNANP (SEQ ID NO: 231).
  • “repeat” in reference to sequence A refers to sequence A being present once, and three repeats of sequence A refers to sequence A being present three times.
  • Multimerization region refers to a region that directs assembly of multimers into a complex, where each multimer comprises a polypeptide associated with the multimerization region.
  • N-terminal domain As used herein, the term “N-terminal domain” refers to a region of a CSP polypeptide that corresponds to amino acids 19-104 of wild-type CSP sequence (SEQ ID NO: 1).
  • N-terminal start region As used herein, the term “N-terminal start region” refers to a region of a CSP polypeptide that corresponds to amino acids 19-31 of wild-type CSP sequence (SEQ ID NO:1).
  • N-terminal end region refers to a region of a CSP polypeptide that corresponds to amino acids 81-92 of wild-type CSP sequence (SEQ ID NO: 1).
  • N-terminal region refers to a region of a CSP polypeptide that corresponds to amino acids 19-80 of wild-type CSP sequence (SEQ ID NO: 1).
  • R1 refers to a region of a CSP polypeptide that corresponds to amino acids 93-97 of wild-type CSP sequence (SEQ ID NO: 1).
  • RNA lipid nanoparticle refers to a nanoparticle comprising at least one lipid and RNA molecule(s), e.g., one or more polyribonucleotides as provided herein.
  • an RNA lipid nanoparticle comprises at least one cationic amino lipid.
  • an RNA lipid nanoparticle comprises at least one cationic amino lipid, at least one helper lipid, and at least one polymer- conjugated lipid (e.g., PEG-conjugated lipid).
  • RNA lipid nanoparticles as described herein can have an average size (e.g., Z-average) of about 100 nm to 1000 nm, or about 200 nm to 900 nm, or about 200 nm to 800 nm, or about 250 nm to about 700 nm.
  • Z-average average size
  • RNA lipid nanoparticles can have a particle size (e.g., Z-average) of about 30 nm to about 200 nm, or about 30 nm to about 150 nm, about 40 nm to about 150 nm, about 50 nm to about 150 nm, about 60 nm to about 130 nm, about 70 nm to about 110 nm, about 70 nm to about 100 nm, about 80 nm to about 100 nm, about 90 nm to about 100 nm, about 70 to about 90 nm, about 80 nm to about 90 nm, or about 70 nm to about 80 nm.
  • a particle size e.g., Z-average
  • an average size of lipid nanoparticles is determined by measuring the average particle diameter.
  • RNA lipid nanoparticles may be prepared by mixing lipids with RNA molecules described herein.
  • Neutralization refers to an event in which binding agents such as antibodies bind to a biological active site of a parasite such as a receptor binding protein, thereby inhibiting the parasitic infection of cells. In some embodiments, the term “neutralization” refers to an event in which binding agents eliminate or significantly reduce ability of infecting cells.
  • nucleic acid refers to a polymer of at least 10-nucleotides or more.
  • a nucleic acid is or comprises DNA.
  • a nucleic acid is or comprises RNA.
  • a nucleic acid is or comprises polypeptide nucleic acid (PNA).
  • PNA polypeptide nucleic acid
  • a nucleic acid is or comprises a single stranded nucleic acid.
  • a nucleic acid is or comprises a double-stranded nucleic acid.
  • a nucleic acid comprises both single and double-stranded portions.
  • a nucleic acid comprises a backbone that comprises one or more phosphodiester linkages. In some embodiments, a nucleic acid comprises a backbone that comprises both phosphodiester and non-phosphodiester linkages. For example, in some embodiments, a nucleic acid may comprise a backbone that comprises one or more phosphorothioate or 5'-N-phosphoramidite linkages and/or one or more polypeptide bonds, e.g., as in a “polypeptide nucleic acid”.
  • a nucleic acid comprises one or more, or all, natural residues (e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uracil). In some embodiments, a nucleic acid comprises on or more, or all, non- natural residues.
  • natural residues e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uracil.
  • a non-natural residue comprises a nucleoside analog (e.g., 2- aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3 -methyl adenosine, 5-methylcytidine, C-5 propynyl- cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl- uridine, C5 -propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8- oxoadenosine, 8-oxoguanosine, 6-O-methylguanine, 2-thiocytidine, methylated bases, intercalated bases, and combinations thereof).
  • a non-natural residue comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) as compared to those in natural residues.
  • a nucleic acid has a nucleotide sequence that encodes a functional gene product such as an RNA or polypeptide.
  • a nucleic acid has a nucleotide sequence that comprises one or more introns.
  • a nucleic acid may be prepared by isolation from a natural source, enzymatic synthesis (e.g., by polymerization based on a complementary template, e.g., in vivo or in vitro), reproduction in a recombinant cell or system, or chemical synthesis.
  • a nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, or 20,000 or more residues or nucleotides long.
  • compositions which achieves a desired reaction or a desired effect alone or together with further doses.
  • a desired reaction in some embodiments relates to inhibition of the course of the disease (e.g., malaria). In some embodiments, such inhibition may comprise slowing down the progress of a disease (e.g., malaria) and/or interrupting or reversing the progress of the disease (e.g., malaria).
  • a desired reaction in a treatment of a disease may be or comprise delay or prevention of the onset of a disease (e.g., malaria) or a condition (e.g., a malaria associated condition).
  • an effective amount of a composition (e.g., a pharmaceutical composition) described herein will depend, for example, on disease (e.g., malaria) or a condition (e.g., a malaria associated condition) to be treated, the severity of such a disease (e.g., malaria) or a condition (e.g., a malaria associated condition), individual parameters of the patient, including, e.g., age, physiological condition, size and weight, the duration of treatment, the type of an accompanying therapy (if present), the specific route of administration and similar factors. Accordingly, doses of a composition (e.g., a pharmaceutical composition) described herein may depend on various of such parameters.
  • Polypeptide refers to a polymeric chain of amino acids.
  • a polypeptide has an amino acid sequence that occurs in nature.
  • a polypeptide has an amino acid sequence that does not occur in nature.
  • a polypeptide has an amino acid sequence that is engineered in that it is designed and/or produced through action of the hand of man.
  • a polypeptide may comprise or consist of natural amino acids, non-natural amino acids, or both.
  • a polypeptide may comprise or consist of only natural amino acids or only non-natural amino acids.
  • a polypeptide may comprise D-amino acids, L-amino acids, or both.
  • a polypeptide may comprise only D-amino acids.
  • a polypeptide may comprise only L-amino acids.
  • a polypeptide may include one or more pendant groups or other modifications, e.g., modifying or attached to one or more amino acid side chains, at the polypeptide’s N-terminus, at the polypeptide’s C-terminus, or any combination thereof.
  • such pendant groups or modifications comprise acetylation, amidation, lipidation, methylation, pegylation, etc., including combinations thereof.
  • a polypeptide may be cyclic, and/or may comprise a cyclic portion. In some embodiments, a polypeptide is not cyclic and/or does not comprise any cyclic portion. In some embodiments, a polypeptide is linear. In some embodiments, a polypeptide may be or comprise a stapled polypeptide.
  • polypeptide may be appended to a name of a reference polypeptide, activity, or structure; in such instances it is used herein to refer to polypeptide that share the relevant activity or structure and thus can be considered to be members of the same class or family of polypeptide.
  • the present specification provides and/or those skilled in the art will be aware of exemplary polypeptide within the class whose amino acid sequences and/or functions are known; in some embodiments, such exemplary polypeptide are reference polypeptide for the polypeptide class or family.
  • a member of a polypeptide class or family shows significant sequence homology or identity with, shares a common sequence motif (e.g., a characteristic sequence element) with, and/or shares a common activity (in some embodiments at a comparable level or within a designated range) with a reference polypeptide of the class; in some embodiments with all polypeptide within the class).
  • a common sequence motif e.g., a characteristic sequence element
  • shares a common activity in some embodiments at a comparable level or within a designated range
  • a member polypeptide shows an overall degree of sequence homology or identity with a reference polypeptide that is at least about 30-40%, and is often greater than about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more and/or includes at least one region (e.g., a conserved region that may in some embodiments be or comprise a characteristic sequence element) that shows very high sequence identity, often greater than 90% or even 95%, 96%, 97%, 98%, or 99%.
  • a conserved region that may in some embodiments be or comprise a characteristic sequence element
  • a conserved region usually encompasses at least 3-4 and often up to 35 or more amino acids; in some embodiments, a conserved region encompasses at least one stretch of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 or more contiguous amino acids.
  • a relevant polypeptide may comprise or consist of a fragment of a parent polypeptide.
  • a polypeptide is a Plasmodium polypeptide construct described herein.
  • a Plasmodium polypeptide construct is a polypeptide that includes one or more Plasmodium proteins, or one or more portions thereof.
  • a Plasmodium polypeptide construct described herein includes at least one region of Plasmodium CSP or a portion thereof.
  • a Plasmodium polypeptide construct additionally includes one or more additional amino acid sequences, such as a secretory signal (e.g., a heterologous secretory signal), a transmembrane region (e.g., a heterologous transmembrane region), a helper antigen, a multimerization region, and/or a linker, as described herein.
  • a secretory signal e.g., a heterologous secretory signal
  • a transmembrane region e.g., a heterologous transmembrane region
  • helper antigen e.g., a multimerization region, and/or a linker, as described herein.
  • Prevent As used herein, the term “prevent” or “prevention” when used in connection with the occurrence of a disease, disorder, and/or condition, refers to reducing the risk of developing the disease, disorder and/or condition and/or to delaying onset of one or more characteristics or symptoms of the disease, disorder or condition. Prevention may be considered complete when onset of a disease, disorder or condition has been delayed for a predefined period of time. In some embodiments, prevention refers to reducing the risk of developing clinical malaria. [0318] Reference: As used herein, the term “reference” describes a standard or control relative to which a comparison is performed.
  • an agent, animal, individual, population, sample, sequence or value of interest is compared with a reference or control agent, animal, individual, population, sample, sequence or value.
  • a reference or control is tested and/or determined substantially simultaneously with the testing or determination of interest.
  • a reference or control is a historical reference or control, optionally embodied in a tangible medium.
  • a reference or control is determined or characterized under comparable conditions or circumstances to those under assessment.
  • RNA Ribonucleic acid
  • RNA Polyribonucleotide
  • RNA Ribonucleic acid
  • polyribonucleotide refers to a polymer of ribonucleotides.
  • an RNA is single stranded.
  • an RNA is double stranded.
  • an RNA comprises both single and double stranded portions.
  • an RNA can comprise a backbone structure as described in the definition of “Nucleic acid / Polynucleotide” above.
  • An RNA can be a regulatory RNA (e.g., siRNA, microRNA, etc.), or a messenger RNA (mRNA).
  • an RNA is a mRNA. In some embodiments, where an RNA is a mRNA, an RNA typically comprises at its 3' end a poly(A) region. In some embodiments, where an RNA is a mRNA, an RNA typically comprises at its 5' end an art-recognized cap structure, e.g., for recognizing and attachment of a mRNA to a ribosome to initiate translation. In some embodiments, an RNA is a synthetic RNA. Synthetic RNAs include RNAs that are synthesized in vitro (e.g., by enzymatic synthesis methods and/or by chemical synthesis methods).
  • a polyribonucleotide encodes a polypeptide, which is preferably is a Plasmodium polypeptide construct.
  • Ribonucleotide encompasses unmodified ribonucleotides and modified ribonucleotides.
  • unmodified ribonucleotides include the purine bases adenine (A) and guanine (G), and the pyrimidine bases cytosine (C) and uracil (U).
  • Modified ribonucleotides may include one or more modifications including, but not limited to, for example, (a) end modifications, e.g., 5' end modifications (e.g., phosphorylation, dephosphorylation, conjugation, inverted linkages, etc.), 3' end modifications (e.g., conjugation, inverted linkages, etc.), (b) base modifications, e.g. , replacement with modified bases, stabilizing bases, destabilizing bases, or bases that base pair with an expanded repertoire of partners, or conjugated bases, (c) sugar modifications (e.g., at the 2' position or 4' position) or replacement of the sugar, and (d) internucleoside linkage modifications, including modification or replacement of the phosphodiester linkages.
  • end modifications e.g., 5' end modifications (e.g., phosphorylation, dephosphorylation, conjugation, inverted linkages, etc.), 3' end modifications (e.g., conjugation, inverted linkages, etc.)
  • base modifications
  • ribonucleotide also encompasses ribonucleotide triphosphates including modified and non-modified ribonucleotide triphosphates.
  • Secretory signal refers to an amino acid sequence motif that targets associated polypeptide for translocation to a secretory pathway.
  • Subject refers to an organism to be administered with a composition described herein, e.g., for experimental, diagnostic, prophylactic, and/or therapeutic purposes. Typical subjects include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, domestic pets, etc.) and humans.
  • a subject is a human subject.
  • a subject is suffering from a disease, disorder, or condition (e.g., malaria and/or a malaria-associated condition).
  • a subject is susceptible to a disease, disorder, or condition (e.g., malaria and/or a malaria-associated condition).
  • a subject displays one or more symptoms or characteristics of a disease, disorder, or condition (e.g., malaria and/or a malaria-associated condition).
  • a subject displays one or more non- specific symptoms of a disease, disorder, or condition (e.g., malaria and/or a malaria-associated condition).
  • a subject does not display any symptom or characteristic of a disease, disorder, or condition (e.g., malaria and/or a malaria-associated condition).
  • a subject is someone with one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition (e.g., malaria and/or a malaria- associated condition).
  • a subject is a patient.
  • a subject is an individual to whom diagnosis and/or therapy is and/or has been administered.
  • Suffering from An individual who is “suffering from” a disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition) has been diagnosed with and/or displays one or more symptoms of a disease, disorder, and/or condition.
  • Susceptible to An individual who is “susceptible to” a disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition) is one who has a higher risk of developing the disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition) than does a member of the general public.
  • an individual who is susceptible to a disease, disorder and/or condition may not have been diagnosed with the disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition).
  • an individual who is susceptible to a disease, disorder, and/or condition e.g., malaria and/or a malaria-associated condition
  • an individual who is susceptible to a disease, disorder, and/or condition may not exhibit symptoms of the disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition).
  • an individual who is susceptible to a disease, disorder, and/or condition e.g., malaria and/or a malaria-associated condition
  • will develop the disease, disorder, and/or condition e.g., malaria and/or a malaria- associated condition).
  • an individual who is susceptible to a disease, disorder, and/or condition will not develop the disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition).
  • therapy refers to an administration or delivery of an agent or intervention that has a therapeutic effect and/or elicits a desired biological and/or pharmacological effect (e.g., has been demonstrated to be statistically likely to have such effect when administered to a relevant population).
  • a therapeutic agent or therapy is any substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition).
  • a therapeutic agent or therapy is a medical intervention that can be performed to alleviate, relieve, inhibit, present, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition.
  • Transmembrane region refers to a region of a polypeptide that spans a biological membrane, such as the plasma membrane of a cell.
  • Treat refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of one or more symptoms or features of a disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition).
  • Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition).
  • treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and/or condition (e.g., malaria and/or a malaria-associated condition), for example for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and/or condition.
  • treatment may be administered to a subject at a later-stage of disease, disorder, and/or condition (e.g., malaria and/or a malaria- associated condition).
  • variant refers to a molecule that shows significant structural (e.g., primary or secondary) identity with a reference molecule but differs structurally from the reference molecule.
  • a variant polypeptide or nucleic acid may differ from a reference polypeptide or nucleic acid as a result of one or more differences in amino acid or nucleotide sequence and/or one or more differences in chemical moieties (e.g., carbohydrates, lipids, phosphate groups) that are covalently components of the polypeptide or nucleic acid (e.g., that are attached to the polypeptide or nucleic acid backbone).
  • moieties e.g., carbohydrates, lipids, phosphate groups
  • Malaria is a mosquito-borne infectious disease caused by single-celled eukaryotic Plasmodium parasites that are transmitted by the bite of Anopheles spp. mosquitoes (Phillips, M., et al. Malaria. Nat Rev Dis Primers 3, 17050, 2017, which is incorporated herein by reference in its entirety).
  • Mosquitoes that transmit malaria must have been infected through a previous blood meal taken from an infected subject (e.g., a human). When a mosquito bites an infected subject a small amount of blood is taken in containing malaria parasites. The infected mosquito can then subsequently bite a non-infected subject, infecting the subject.
  • RTS,S/AS01 is an adjuvanted protein subunit vaccine that consists of a portion of the major repeat region and the C- terminus of CSP from Plasmodium falciparum fused to the Hepatitis B surface antigen (HBsAg).
  • the vaccine is a mix of this PfCSP-HBsAg compound with HBsAg that forms virus-like particles (RTS,S/AS01; MosquirixTM).
  • RTS,S is administered according to a regimen that requires four doses: an initial 3-dose schedule given at least 1 month apart, and a 4th dose 15-18 months after dose 3 (see, for example, Vandoolaeghe & Schuerman Expert Rev Vaccines.
  • RTS,S protects approximately 30% to 50% of children from clinical disease over 18 months.
  • RTS,S has been reported to induce protective antibody and CD4+ T-cell responses, but only negligible CD8+ T cell responses (see, for example, Moris et al. Hum Vaccin Immunother 14:17, 2018, which is incorporated herein by reference in its entirety).
  • CSP circumsporozoite protein
  • CSP precipitation reaction due to the density and close proximity of neighboring CSPs on the surface of the parasite coupled with the bi-valency of antibodies, binding of antibodies to CSP can produce a phenomenon referred to as CSP precipitation reaction, whereby antibodies can crosslink neighboring CSP and cause them to precipitate and shed from the parasite surface, leaving a trail of precipitated antibody bound CSP that the parasite can replace through its normal CSP translocation process (Livingstone et al., Sci Rep 11, 5318 (2021); Steward et al., J Protozool. 1991 Jul- Aug; 38(4):411-21, each of which is incorporated herein by reference in its entirety).
  • sporozoites When moving from an inoculation site in the skin to the liver, sporozoites traverse host cells (Mota et al., Science 2001 Jan 5;291(5501):141-4). Sporozoites traverse different types of host cells at the dermis, including fibroblasts and phagocytes (Amino et al., Cell Host Microbe.2008 Feb 14;3(2):88-96, which is incorporated herein by reference in its entirety), and the liver sinusoidal barrier, containing liver endothelial cells and Kupffer cells (Frevert et al., PLoS Biol 3(6): e192.2005, which is incorporated herein by reference in its entirety) and sinusoidal endothelial cells (Tavares et al., J Exp Med 2013 May 6;210(5):905-15, which is incorporated herein by reference in its entirety), in order to gain access to hepatocytes.
  • HSPGs low-sulfated heparin sulfate proteoglycans
  • High-sulfated HSPGs Cell traversal was first observed as non-phagocytic entry of P. berghei sporozoites into macrophages followed by “escape” from these cells (Vanderberg et al., J. Euk. Microbiol.37:528-536, 1990, which is incorporated herein by reference in its entirety).
  • HGF hepatocyte growth factor
  • GPDH glyceraldehyde 3- phosphate dehydrogenase
  • SPECT2 sporozoite microneme protein essential for cell traversal 2
  • PBP1 perforin-like protein 1
  • yoelii has been shown to play a role in cell traversal. Although this protein is not required for hepatocyte entry, it plays a role in egress from transient vacuoles during traversal (Risco- Castillo et al., 2015, each of which is incorporated herein by reference in its entirety). Thus, sporozoites that infect rodents can traverse host cells by generating a vacuole at the entry step and use a perforin-like protein (e.g., SPECT2/PLP1) to escape from this compartment and/or a host cell, during cell exit.
  • SPECT2/PLP1 perforin-like protein
  • sporozoites Once sporozoites have invaded liver cells, they differentiate into merozoites, a replicative form of the parasite capable of lysing hepatocytes after multiple rounds of replication. Within a few days, a few hundred sporozoites can become hundreds of thousands of merozoites. When infected liver cells rupture, they release the merozoites into the bloodstream, where they invade red blood cells and begin the asexual reproductive stage, which is the symptomatic stage of the disease. Within a small number of days, millions of merozoites can be present in blood. [0341] Malaria symptoms typically develop 4-8 days after initial red blood cell invasion.
  • Replication cycle of merozoites within the red blood cells continues for 36-72 hours, until hemolysis, releasing the merozoites for another round of red blood cell infection.
  • fever occurs every 36–72 hours, when infected red blood cells lyse and release endotoxins en masse.
  • Plasmodium spp. parasites gain entry into red blood cells through specific ligand–receptor interactions mediated by proteins on the surface of the parasite that interact with receptors on the host erythrocyte (mature red blood cell) or reticulocyte (immature red blood cell), whereas P. falciparum can invade and replicate in erythrocytes and reticulocytes, P.
  • vivax and other species predominantly invade reticulocytes, which are less abundant than erythrocytes. Most of the erythrocyte-binding proteins or reticulocyte-binding proteins that have been associated with invasion are redundant or are expressed as a family of variant forms; however, for P. falciparum, two essential red blood cell receptors (basigin and complement decay-accelerating factor (also known as CD55)) have been identified.
  • Plasmodium vivax and Plasmodium ovale can also enter a dormant state in the liver, the hypnozoite.
  • Merozoites released from red blood cells can invade other red blood cells and continue to replicate, or in some cases, they differentiate into male or female gametocytes.
  • Gametocytes concentrate in skin capillaries and are then taken up by the mosquito vector in another blood meal.
  • each male gametocyte produces eight microgametes after three rounds of mitosis; the female gametocyte matures into a macrogamete.
  • Male microgametes are motile forms with flagellae and seek the female macrogamete.
  • the male and female gametocytes fuse, forming a diploid zygote, which elongates into an ookinete; this motile form secretes a chitinase in order to enter the peritrophic membrane and traverse the midgut epithelium to the basal lateral side of the midgut, establishing itself in the basal lamina as an oocyst.
  • Oocysts mature over 14-15 days, undergoing cycles of replication to form sporozoites that are ultimately liberated into the hemocoel, an environment rich in sugars and substrates beneficial to the parasite’s survival.
  • Thousands of sporozoites can form from a single oocyst and become randomly distributed throughout the hemocoel. These sporozoites are motile and rapidly destroy the hemolymph, with only approximately 20% successfully invading the salivary gland. Following invasion of the salivary gland, sporozoites are re-programmed via an unknown mechanism to prepare for liver invasion.
  • Plasmodium parasites are haploid throughout their life cycle.
  • the genomes of different species range from 20 to 35 megabases, contain 1 4 chromosomes, a circular plastid genome of approximately 35 kilobases, and multiple copies of a 6 kilobase mitochondrial DNA. Comparison of genomes from different species showed that homologous genes are often found in synthetic blocks arranged in different orders among different chromosomes.
  • the adenine-thymine (AT) content of Plasmodium spp. can also be very different, e.g., ⁇ 80% AT in P. falciparum, P. vraowi, and P.
  • AT content is often higher in introns and intergenic noncoding regions than in protein- coding exons, with an average of 80.6% AT for the whole P. falciparum genome versus 86.5% for noncoding sequences.
  • the high AT content of P. falciparum reflects large numbers of low-complexity regions, simple sequence repeats, and microsatellites, as well as a highly skewed codon usage bias.
  • Polymorphisms of AT-rich repeats provide abundant markers for linkage mapping of drug resistance genes and for tracing the evolution and structure of parasite populations.
  • Malaria parasite genomes carry multigene families that serve important roles in parasite interactions with their hosts, including, for example, antigenic variation, signaling, protein trafficking, and adhesion.
  • genes encoding P. falciparum erythrocyte membrane protein 1 (PfEMP1) have been studied most extensively.
  • Each individual P. falciparum parasite carries a unique set of 50 to 150 copies of the var gene in its genome, where switches of gene expression can produce antigenic variation.
  • PfEMP1 plays an important role in the pathogenesis of clinical developments such as in cerebral and placental malaria, in which it mediates the cytoadherence of infected red blood cells (iRBCs; infected erythrocytes) in the deep tissues.
  • iRBCs infected red blood cells
  • Different PfEMP1 molecules bind to various host molecules, including ⁇ 2-macroglobulin, CD36, chondroitin sulfate A (CSA), complement 1q, CR1, E-selectins and P-selectins, endothelial protein C receptor (EPCR), heparan sulfate, ICAM1, IgM, IgG, PECAM1, thrombospondin (TSP), and VCAM1.
  • CSA chondroitin sulfate A
  • EPCR endothelial protein C receptor
  • ICAM1 heparan sulfate
  • IgM IgM
  • IgG IgG
  • PECAM1 thrombospondin
  • VCAM1 thrombospondin
  • An additional, exemplary polymorphic gene family comprises a group of 14 genes encoding proteins with six cysteines (6-Cys).
  • falciparum is largely conserved in central regions but extensively polymorphic is both length and sequence near the telomeres. Much of the subtelomeric variation was explained by recombination within blocks of repetitive sequences and families of genes. [0353]
  • the frequency of simple sequence repeats (microsatellites) in P. falciparum is estimated to be approximately one polymorphic microsatellite per kb DNA. Without wishing to be bound by any one theory, this high rate may reflect the AT-rich nature of the genome. Microsatellites seem to be less frequent in other Plasmodium species that have genomes with lower AT contents.
  • Plasmodium parasites are known to express various proteins at different stages of their lifecycles. Exemplary Plasmodium proteins are described below, and exemplary amino acid sequences are provided in Table 2.
  • Circumsporozoite protein is a multifunctional protein that is involved in Plasmodium life cycle, as it is required for the formation of sporozoites in the mosquito midgut, the release of sporozoites from the oocyst, invasion of salivary glands, attachment of sporozoites to hepatocytes in the liver, and sporozoite invasion of hepatocytes (see, e.g., Zhao et al. (2016) PLoS ONE 11(8): e0161607, which is incorporated herein by reference in its entirety).
  • CSP CSP is present in all Plasmodium species, and although variation exists in the amino acid sequence across species, the overall domain structure of a central repeat region and nonrepeat flanking regions is well conserved (see, e.g., Zhao et al. (2016) PLoS ONE 11(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 known (see, e.g., 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), and Table 1 includes exemplary sequences for CSP P. falciparum isolates from Asia, South America and Africa. Table 1: Exemplary Sequences Encoding CSP P. falciparum isolates from Asia, South America and Africa
  • An exemplary wild-type CSP polypeptide amino sequence from Plasmodium falciparum isolate 3D7 is presented in Table 2 as SEQ ID NO: 1, and includes the following: a secretory signal (amino acids 1-18); an N- terminal domain (amino acids 19-104); a junction region (amino acids 93-104), a central domain (amino acids 105- 272); and a C-terminal domain (amino acids 273-397).
  • the N-terminal domain includes an N-terminal region (amino acids 19-80); an N-terminal end region (amino acids 81-92); and a junction region (amino acids 93-104).
  • the junction region includes an R1 region (amino acids 93-97) and a junction (SEQ ID NO: 277) at positions 98-104.
  • the central domain includes a minor repeat region (amino acids 105-128) and a major repeat region (amino acids 129-272).
  • the minor repeat region includes three repeats of the amino acid sequence NANPNVDP (SEQ ID NO: 223).
  • the major repeat region includes 35 repeats of the amino acid sequence NANP (SEQ ID NO: 230), wherein 35 repeats of the amino acid sequence NANP (SEQ ID NO: 230) are separated into two contiguous stretches, and wherein one stretch includes 17 repeats of the amino acid sequence NANP (SEQ ID NO: 230) and one includes 18 repeats of the amino acid sequence NANP (SEQ ID NO: 230) which flank an amino acid sequence of NVDP (SEQ ID NO: 229).
  • the major repeat region includes the amino acid sequences NPNANP (SEQ ID NO: 231) and NANPNA (SEQ ID NO: 232).
  • the C-terminal domain includes a C-terminal region (amino acids 273-375), a serine-valine (amino acids 376-377), and a transmembrane domain (amino acids 378-397).
  • the C-terminal region includes a Th2R region (amino acids 314-327) and a Th3R region (amino acids 352-363).
  • Exemplary CSP amino acid sequence is provided in Table 2.
  • UIS3 was shown to interact with liver fatty acid- binding protein (L-FABP) and be involved in fatty acid and/or lipid import during phases of Plasmodium growth (see, e.g., 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 its entirety).
  • L-FABP liver fatty acid- binding protein
  • the Plasmodium relies on host fatty acids for rapid synthesis of its membranes (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 in the PVM provides Plasmodium a method to import essential fatty acids and/or lipids during rapid sporozoites growth phases (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-deficient Plasmodium berghei are also unable to develop into mature liver schizonts and therefore abort malaria infection within the liver itself (see, e.g., Mueller et al., Nature. 2005 Jan 13;433(7022):164-7, which is incorporated herein by reference in its entirety). Further, it was previously demonstrated that UIS3 derived from Plasmodium berghei and UIS3 derived from Plasmodium falciparum exhibited a low (i.e. 34%) amino acid sequence identity (see, e.g., Mueller et al., Nature. 2005 Jan 13;433(7022):164-7, which is incorporated herein by reference in its entirety).
  • Plasmodium UIS3 sequences are known (see, e.g., UniProt accession number A0A509ARS3, A0A1C6YLP3, Q8IEU1, A0A384KLI1, A0A1G4H423, A0A077YB01, Q9NFU4, each of which is incorporated herein by reference in its entirety). Exemplary UIS3 amino acid sequence is provided in Table 2. [0361] Plasmodium falciparum early transcribed membrane protein 10.3 (ETRAMP10.3) is an approximately 10 kDa protein and member of the early transcribed membrane proteins multigene family, a family which is conserved across Plasmodium species and includes proteins located in the parasitophorous vacuole.
  • ETRAMP10.3 is one example, which is expressed in both liver and blood stage P. falciparum parasites. ETRAMP10.3 transcription has been found to peak during the transition from ring to trophozoite stages of P. falciparum blood stage infection in a human host. ETRAMP10.3 localizes to the parasitophorous vacuole and is exported to a host erythrocyte during blood stage infection.
  • ETRAMP10.3 is sometimes referred to as Upregulated in Infectious Sporozoites gene 4 (UIS4)
  • UIS4 Upregulated in Infectious Sporozoites gene 4
  • ETRAMP10.3 is not a functional ortholog of UIS4 and may play a different biological role. Although the biological function of ETRAMP10.3 has not yet been completely resolved, localization to vesicular structures in the host erythrocyte suggests a role in host-parasite interaction or in remodeling of infected erythrocyte. ETRAMP10.3 appears to play a key role in the Plasmodium life cycle. When ETRAMP10.3 is deleted, the deletion can lead to the disruption of liver-stage development in mice and asexual blood stage progression.
  • LISP-1 Liver specific protein 1
  • PVM parasitophorous vacuolar membrane
  • Intracellular Plasmodium deficient in LISP-1 develop into hepatic merozoites and display normal infectivity to erythrocytes (see, e.g., Ishino et al., Cell Microbiol. 2009 Sep; 11(9): 1329–1339, which is incorporated herein by reference in its entirety).
  • LISP1-deficient liver-stage Plasmodium do not rupture PVM and remain trapped inside hepatocytes (see, e.g., Ishino et al., Cell Microbiol.2009 Sep; 11(9): 1329–1339, which is incorporated herein by reference in its entirety).
  • Plasmodium LISP-1 sequences are known (see, e.g., UniProt accession number A0A2I0C2X6, Q8ILR5, each of which is incorporated herein by reference in its entirety). Exemplary LISP-1 amino acid sequence is provided in Table 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, which is incorporated herein by reference in its entirety).
  • LISP-2 was shown to be expressed by liver stages Plasmodium, exported to hepatocytes, and be distributed throughout the host cell, including the nucleus (see, e.g., Orito et al., Mol Microbiol. 2013 Jan;87(1):66-79, which is incorporated herein by reference in its entirety).
  • Intracellular Plasmodium deficient in LISP2 do not mature effectively during merozoites development (see, e.g., Orito et al., Mol Microbiol.2013 Jan;87(1):66-79, which is incorporated herein by reference in its entirety).
  • Plasmodium LISP-2 sequences are known (see, e.g., UniProt accession number A0A2I0BZR4, Q8I1X6, Q9U0D4, each of which is incorporated herein by reference in its entirety).
  • Exemplary LISP-2 amino acid sequence is provided in Table 2.
  • Thrombospondin-related adhesion protein contains an N-terminal domain that is commonly referred to as von Willebrand factor A domain, although it is most similar to an integrin I domain because it contains a metal ion-dependent adhesion site (MIDAS) with a bound Mg 2+ ion that is required for sporozoite motility in vitro and infection in vivo (see, e.g., Lu et al., PLoS One.2020; 15(1): e0216260, which is incorporated herein by reference in its entirety).
  • MIDAS metal ion-dependent adhesion site
  • the I domain is inserted in an extensible ⁇ -ribbon and followed by a thrombospondin repeat (TSR) domain, a proline-rich segment at the C-terminus, a single-pass transmembrane domain, and a cytoplasmic domain (see, e.g., Lu et al., PLoS One.2020; 15(1): e0216260, which is incorporated herein by reference in its entirety).
  • TSR thrombospondin repeat
  • TRAP is stored in the micronemes and becomes surface exposed at the sporozoite anterior tip when parasite comes in contact with host cells (Akhouri et al., Malar J. 2008 Apr 22;7:63. doi: 10.1186/1475-2875-7-63, which is incorporated herein by reference in its entirety). TRAP also plays an important role in liver cell invasion of sporozoites by helping sporozoites in gliding motility and in recognition of host receptors on the mosquito salivary gland and hepatocytes (Akhouri et al., Malar J. 2008 Apr 22;7:63. doi: 10.1186/1475-2875-7-63, which is incorporated herein by reference in its entirety).
  • Plasmodium TRAP sequences are known (see, e.g., 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 sequence is provided in Table 2.
  • LSAP-1 Liver-stage-associated protein
  • LSAP-1 has been shown to be found mainly at the periphery of the intracellular hepatic parasite throughout its development, but not in blood stage parasites and possibly in minor quantities in salivary gland sporozoites (see, e.g., Siau et al., PLoS Pathog.2008 Aug 8;4(8):e1000121, which is incorporated herein by reference in its entirety).
  • LSAP-1 is among the most abundant transcripts in the salivary gland transcriptome but has not been detected in proteomic surveys of sporozoites. Rather, expression has only been detected only in liver stages (see, e.g., Siau et al., PLoS Pathog.
  • Plasmodium LSAP-1 sequences are known (see, e.g., UniProt accession number Q8I632, W7JR53, each of which is incorporated herein by reference in its entirety). Exemplary LSAP-1 amino acid sequence is provided in Table 2. [0375] Like LSAP-1, LSAP-2 is also among the most abundant transcripts in the salivary gland transcriptome but has not been detected in proteomic surveys of sporozoites. LSAP-2 has shown some efficacy as a vaccine when combined with other antigens.
  • Plasmodium LSAP-2 sequences are known (see, e.g., UniProt accession number Q8I632, W7JR53, each of which is incorporated herein by reference in its entirety). Exemplary LSAP-2 amino acid sequence is provided in Table 2.
  • LSA-1 Liver-Stage Antigen 1
  • the function of LSA-1 remains currently not known (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vaccine Candidates in Malaria', in A. J. Rodriguez-Morales (ed.), Current Topics in Malaria, IntechOpen, London.10.5772/65592, each of which is incorporated herein by reference in its entirety).
  • the function of LSA-1 remains currently not known (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vaccine Candidates in Malaria', in A. J.
  • LSA-1 is a 230 kDa preerythrocytic stage protein containing a large central region consisting of over eighty 17 amino acid residue repeat units flanked by highly conserved C- and N-terminal regions (Richie, T.L. and Parekh, F.K. (2009) Malaria, which is incorporated herein by reference in its entirety).
  • Vaccines for Biodefense and Emerging and Neglected Diseases Barrett, A.D.T. and Stanberry L.R., eds
  • LSA1 is expressed only by liver stage Plasmodium and not by sporozoites (Richie, T.L. and Parekh, F.K. (2009) Malaria, which is incorporated herein by reference in its entirety).
  • Vaccines for Biodefense and Emerging and Neglected Diseases Barrett, A.D.T. and Stanberry L.R., eds
  • pp. 1309– 1364 Elsevier, which is incorporated herein by reference in its entirety.
  • the repeat region results in significant variation of the protein between strains of Plasmodium falciparum (see, e.g., Tucker, K.
  • Plasmodium LSA-1 sequences are known (see, e.g., UniProt accession number Q25886, Q25887, Q25893, Q26028, Q9GTX5, O96125, each of which is incorporated herein by reference in its entirety).
  • Exemplary LSA-1 amino acid sequence is provided in Table 2.
  • LSA-3 Liver stage antigen 3 is a 200-kDa protein that is composed of three nonrepeating regions (NR- A, NR-B, and NR-C) flanking two short repeat regions and one long repeat region (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vaccine Candidates in Malaria', in A. J. Rodriguez-Morales (ed.), which is incorporated herein by reference in its entirety), Current Topics in Malaria, IntechOpen, London. 10.5772/65592, which is incorporated herein by reference in its entirety).
  • the nonrepeat 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 A. J. Rodriguez-Morales (ed.), Current Topics in Malaria, IntechOpen, London. 10.5772/65592, which is incorporated herein by reference in its entirety).
  • the most significant variation is in the repeating regions due to organization and number of repeating subunits rather than composition of the repeating regions (see, e.g., Tucker, K. et al., 2016, 'Pre-Erythrocytic Vaccine Candidates in Malaria', in A. J.
  • Plasmodium LSA-3 sequences are known (see, e.g., UniProt accession number C7DU21, C7DU22, C7DU23, C7DU24, C7DU25, C7DU26, C7DU27, C7DU28, C7DU29, C7DU32, C7DU33, C7DU34, C7DU36, C7DU37, C7DU38, C7DU39, C7DU40, Q8I042, Q8I0A5, Q8I0D0, Q8IFR1, Q8IFR2, Q8IFR3, Q8IFR4, Q8IFR5, Q8IFR6, Q8IFR7, Q8IFR8, Q8IFR9, Q8IFS0, Q8IFS1, Q8IFS2, Q8IFS3, Q8IFS4, Q8IFS5, Q8IFS6, Q8IFS7, Q8IFS8, Q8IFS9, Q8IFT0, Q8IFT1, Q8IFT2, Q8IFT3, Q8IFS
  • the present disclosure provides combinations of polyribonucleotides that can be used to express one or more Plasmodium polypeptide constructs encoding at least two Plasmodium polypeptides or antigenic portions thereof, wherein the at least two Plasmodium polypeptides or antigenic portions thereof are expressed during different stages of the Plasmodium life cycle.
  • a combination as described herein comprises one or more polyribonucleotides that encode one or more polypeptides or antigenic portions thereof that are expressed during the Plasmodium sporozoite stage (e.g., shortly after a Plasmodium parasite has entered a subject and before the Plasmodium parasite has infected a hepatocyte), and one or more polypeptides or antigenic portions thereof that are expressed during the Plasmodium liver stage (e.g., after a Plasmodium parasite has entered a hepatocyte).
  • a combination as described herein comprises one or more polyribonucleotides that encode one or more Plasmodium liver stage polypeptides or antigenic portions thereof (e.g., that elicit a T cell response) and one or more polyribonucleotides that encode one or more Plasmodium sporozoite stage (e.g., initial sporozoite stage) polypeptides or antigenic portions thereof (e.g., CSP).
  • one or more Plasmodium liver stage polypeptides or antigenic portions thereof comprises between 2 to 20 liver stage polypeptides or antigenic portions thereof (e.g., antigenic portions that induce a T cell response).
  • a combination as described herein comprises one or more polyribonucleotides that encode one or more Plasmodium T-cell string polypeptide constructs as described herein and one or more polyribonucleotides that encode one or more Plasmodium CSP polypeptide constructs as described herein.
  • a combination as described herein comprises a first polyribonucleotide that encodes a Plasmodium T-cell string polypeptide construct as described herein and a second polyribonucleotide that encodes a Plasmodium CSP polypeptide construct as described herein.
  • a combination as described herein comprises a first polyribonucleotide that encodes a first Plasmodium T-cell string polypeptide construct as described herein, a second polyribonucleotide that encodes a second Plasmodium T-cell string polypeptide construct as described herein, and a third polyribonucleotide that encodes a Plasmodium CSP polypeptide construct as described herein.
  • a combination comprises a first pharmaceutical composition and a second pharmaceutical composition.
  • a first pharmaceutical composition comprises a first polyribonucleotide, wherein the first polyribonucleotide encodes a Plasmodium T-cell string polypeptide.
  • a second pharmaceutical composition comprises a second polyribonucleotide, wherein the first polyribonucleotide encodes a Plasmodium CSP polypeptide.
  • a first pharmaceutical composition and a second pharmaceutical composition are the same pharmaceutical composition.
  • a first pharmaceutical composition and a second pharmaceutical composition are different pharmaceutical compositions.
  • a combination comprises a first pharmaceutical composition, a second pharmaceutical composition, and a third pharmaceutical composition.
  • a first pharmaceutical composition comprises a first polyribonucleotide, wherein the first polyribonucleotide encodes a first Plasmodium T- cell string polypeptide.
  • a second pharmaceutical composition comprises a second polyribonucleotide, wherein the second polyribonucleotide encodes a second Plasmodium T-cell string polypeptide.
  • a third pharmaceutical composition comprises a third polyribonucleotide, wherein the third polyribonucleotide encodes a Plasmodium CSP polypeptide.
  • a first pharmaceutical composition, a second pharmaceutical composition, and a third pharmaceutical composition are the same pharmaceutical composition.
  • a first pharmaceutical composition, a second pharmaceutical composition, and a third pharmaceutical composition are different pharmaceutical compositions.
  • a first pharmaceutical composition and a second pharmaceutical composition are the same pharmaceutical composition.
  • Plasmodium T-cell string polypeptide constructs are provided below.
  • A. Plasmodium T-Cell String Polypeptide Constructs utilizes RNA technologies as a modality to express one or more Plasmodium T-cell string polypeptide constructs (also referred to as “Plasmodium T-cell string polypeptides,” “malaria T-cell string polypeptide constructs” or “malarial T-cell peptide string constructs”).
  • Plasmodium T-cell string polypeptide constructs as described herein can include one or more T-cell antigens from one or more Plasmodium polypeptide, or one or more portions thereof, (e.g., one or more antigenic fragments thereof) as described herein.
  • a Plasmodium T-cell string polypeptide construct as described herein comprises one or more Plasmodium liver stage antigens that elicit a T cell response.
  • a “T-cell antigen” as described herein can induce a T cell response in a subject or model system.
  • a Plasmodium T- cell string polypeptide construct that targets the liver stage of a Plasmodium infection includes polypeptides or antigenic portions thereof that are expected to be both of relatively high abundance in infected hepatocytes and elicit T cell response(s).
  • Polyribonucleotides as described herein encoding Plasmodium T-cell string polypeptide constructs, as well as Plasmodium T-cell string polypeptide constructs described herein are designed to deliver a polypeptide to a subject, and in turn, for protein degradation and processing for presentation within the subject so that the subject raises an immune response (e.g., T cell response(s)).
  • Plasmodium T-cell string polypeptide constructs as described herein include more than one T-cell antigen and/or epitope from Plasmodium liver stage polypeptides or one or more portions thereof.
  • one or more Plasmodium liver stage polypeptides or antigenic portions thereof comprises between 2 to 20 liver stage polypeptides or antigenic portions thereof (e.g., antigenic portions that induce a T cell response).
  • a Plasmodium T-cell string polypeptide construct comprises 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20, preferably 4, 5, 6, 7, 8, 9, 10, 11 or 12 different liver stage polypeptides or antigenic portions thereof (e.g., which each are capable of eliciting a T-cell response).
  • a Plasmodium T-cell string polypeptide construct includes one or more Plasmodium T-cell antigens from CSP, LSA-1 (e.g., LSA-1(a), LSA-1(b)), TRAP, LSAP2, UIS3, ETRAMP10.3, LISP-1, LISP-2, LSA-3, EXP1, LSAP1 and/or polypeptide regions or portions thereof (e.g., one or more antigenic fragments thereof).
  • LSA-1 e.g., LSA-1(a), LSA-1(b)
  • TRAP e.g., LSA-1(a), LSA-1(b)
  • TRAP e.g., TRAP, LSAP2, UIS3, ETRAMP10.3, LISP-1, LISP-2, LSA-3, EXP1, LSAP1 and/or polypeptide regions or portions thereof (e.g., one or more antigenic fragments thereof).
  • a Plasmodium T-cell string polypeptide construct comprises between about 10 amino acids and about 1200 amino acids, e.g., between about 10 amino acids and about 1100 amino acids, e.g., between about 10 amino acids and about 1000 amino acids, e.g., between about 10 amino acids and about 750 amino acids, e.g., between about 25 amino acids and about 500 amino acids.
  • a Plasmodium T-cell string polypeptide construct comprises about 10, about 15, about 20, about 25, about 50, about 75, about 100, about 150, about 200, about 250, about 300, about 350, about 400, about 450, about 500, about 550, about 600, about 650, about 700, about 750, about 800, about 850, about 900, about 950, about 1000, about 1050, about 1100, about 1150, or about 1200 amino acids.
  • a Plasmodium T-cell string polypeptide construct additionally includes one or more additional amino acid sequences, such as a secretory signal (e.g., a heterologous secretory signal), a transmembrane region (e.g., a heterologous transmembrane region), a trafficking signal, and/or a linker, as described herein.
  • a secretory signal e.g., a heterologous secretory signal
  • a transmembrane region e.g., a heterologous transmembrane region
  • a trafficking signal e.g., a trafficking signal, and/or a linker, as described herein.
  • a T-cell antigen utilized in a Plasmodium T-cell string polypeptide construct described herein includes Plasmodium protein sequences identified and/or characterized by one or more of: - HLA-I or HLA-II binding (e.g., to HLA allele(s) present in a relevant population) - HLA ligandomics data confirmed by mass spectrometry - Relatively high expression - Sequence conservation - Expression during the early liver stage of parasite lifecycle - Localization to the parasitophorous vaculous membrane - Serum reactivity - Immunogenicity (e.g., presence of one or more B-cell and/or T-cell epitopes; evidence of ability to induce sterile protection in model systems including, e.g., humans, non-human primates, and/or mice).
  • - HLA-I or HLA-II binding e.g., to HLA allele(s) present in a relevant population
  • HLA ligandomics data confirmed by mass
  • HLA-I and/or HLA-II binding is experimentally assessed; in some embodiments it is predicted. In some embodiments, predicted HLA-I or HLA-II binding is assessed using an algorithm such as neonmhc 1 and/or neonmhc2, which predict and/or characterize likelihood of MHC class I and MHC class II binding, respectively.
  • an MHC-polypeptide presentation prediction algorithm or MHC-polypeptide presentation predictor is or comprises NetMHCpan or NetMHCIIpan.
  • a hidden Markov model approach may be utilized for MHC-polypeptide presentation prediction and/or characterization.
  • the polypeptide prediction model MARIA may be utilized.
  • NetMHCpan is not utilized to predict or characterize likelihood of MHC binding for polypeptides as described herein.
  • the polypeptide prediction model MARIA may be utilized.
  • NetMHCIIpan is not utilized to predict or characterize likelihood of MHC binding for polypeptides as described herein.
  • an MHC-polypeptide presentation prediction algorithm or MHC-polypeptide presentation predictor is or comprises RECON® (Real-time Epitope Computation for ONcology), which offers high quality MHC-polypeptide presentation prediction based on expression, processing and binding capabilities. See, for example, Abelin et al., Immunity 21:315, 2017; Abelin et al., Immunity 15:766, 2019, each of which is incorporated herein by reference in its entirety.
  • HLA binding and/or ligandomics assessments can consider the geographic region of subjects to be immunized.
  • HLA allelic diversity can be considered.
  • T cell antigens comprise polypeptides (e.g., epitopes) expected or determined, when considered together, to bind to a significant percentage (e.g., at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or more) of HLA alleles expected or known to be present in a relevant region or population.
  • T cell antigens comprise polypeptides expected or determined, when considered together, to bind to the most prevalent (e.g., the 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 most prevalent, or at least 1, 2, 3, 4, or 5 of the 10 most prevalent, etc.) HLA alleles expected or known to be present in a relevant region or population).
  • expression level is experimentally determined (e.g., in a model system or in infected humans). In some embodiments, expression level is a reported level (e.g., in a published or presented report). In some embodiments, expression level is assessed as RNA (e.g., via RNASeq). In some embodiments (and typically preferably), expression levels is assessed as protein.
  • sequence conservation is assessed, for example, using publicly available sequence evaluation software (such as, for example, multiple sequence alignment programs MAFFT, Clustal Omega, etc.). In some embodiments, sequence conservation is determined by consultation with published resources (e.g., sequences). In some embodiments, sequence conservation includes consideration of currently or recently detected strains (e.g., in an active outbreak). [0398] In some embodiments, surface exposure is assessed by reference to publicly available database and/or software.
  • publicly available sequence evaluation software such as, for example, multiple sequence alignment programs MAFFT, Clustal Omega, etc.
  • sequence conservation is determined by consultation with published resources (e.g., sequences).
  • sequence conservation includes consideration of currently or recently detected strains (e.g., in an active outbreak).
  • surface exposure is assessed by reference to publicly available database and/or software.
  • surface exposure is assessed by reference to publicly available data, e.g., as described in Swearingen et al., “Interrogating the Plasmodium Sporozoite Surface: Identification of Surface-Exposed Proteins and Demonstration of Glycosylation on CSP and TRAP by Mass Spectrometry-Based Proteomics” PLoS Pathog (2016), the content of which is incorporated herein by reference for the purposes described herein.
  • serum reactivity is assessed by contacting serum samples from infected individuals with polypeptide including sequences of interest (e.g., as may be displayed via, for example, phage display or polypeptide array, etc.; see, for example, Whittemore et al PlosOne, 2016, which is incorporated herein by reference in its entirety).
  • serum reactivity is assessed by consultation with literature reports and or database data indicating serum-recognized sequences.
  • assessment of immunoreactivity and/or of presence of an epitope may be or comprise consultation with the Immune Epitope Database (IEDB) which those skilled in the art will be aware is a freely available resource funded by NIAID that catalogs experimental data on antibody and T cell epitopes (see iedb.org).
  • IEDB Immune Epitope Database
  • ability to induce sterile protection is assessed, for example, as described in one or more of Schofield et al. “ ⁇ Interferon, CD8+ T cells and antibodies required for immunity to malaria sporozoites” Nature 330, 664–666 (1987); Weiss et al. (1988).
  • CD8+ T cells cytotoxic/suppressors are required for protection in mice immunized with malaria sporozoites” Proc. Natl. Acad. Sci. U.S.A. 85, 573–576; Romero et al. “Cloned cytotoxic T cells recognize an epitope in the circumsporozoite protein and protect against malaria.” Nature 341, 323–326 (1989); Rodrigues et al. (1991) “CD8+ cytolytic T cell clones derived against the Plasmodium yoelii circumsporozoite protein protect against malaria.” Int. Immunol. 3, 579–585; Chakravarty et al.
  • T cell antigens are characterized by dendritic cell presentation which, in turn may be indicative of HLA binding and/or of immunogenicity.
  • dendritic cell presentation e.g., in peripheral lymph nodes, may induce CD8+ T cells that migrate to the liver and, for example, may eliminate parasite-infected hepatocytes. See, for example, Chakravarty et al.
  • Plasmodium T-cell string polypeptide construct described herein includes one or more one or more Plasmodium T-cell antigens.
  • Plasmodium T-cell string polypeptide construct described herein includes one or more Plasmodium T-cell antigens from Plasmodium CSP, LSA-1 (e.g., LSA- 1(a), LSA-1(b)), TRAP, LSAP2, UIS3, ETRAMP10.3, LISP-1, LISP-2, LSA-3, EXP1, LSAP1, or a combination thereof.
  • LSA-1 e.g., LSA- 1(a), LSA-1(b)
  • TRAP e.g., TRAP, LSAP2, UIS3, ETRAMP10.3, LISP-1, LISP-2, LSA-3, EXP1, LSAP1, or a combination thereof.
  • a Plasmodium T-cell string polypeptide construct described herein includes 2 to about 20 Plasmodium T-cell antigens (e.g., about 2 to about 15, about 2 to about 10, about 2 to about 9, about 2 to about 8, about 2 to about 7, about 2 to about 6, or about 2 to about 5 Plasmodium T-cell antigens). In some embodiments, a Plasmodium T-cell string polypeptide construct described herein includes about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 Plasmodium T-cell antigens. In some embodiments, a Plasmodium T-cell string polypeptide construct described herein includes four Plasmodium T-cell antigens.
  • a Plasmodium T-cell string polypeptide construct described herein includes five Plasmodium T-cell antigens. In some embodiments, a Plasmodium T-cell string polypeptide construct described herein includes six Plasmodium T-cell antigens. In some embodiments, a Plasmodium T-cell string polypeptide construct described herein includes seven Plasmodium T-cell antigens. In some embodiments, a Plasmodium T-cell string polypeptide construct described herein includes eight Plasmodium T-cell antigens. In some embodiments, a Plasmodium T-cell string polypeptide construct described herein includes nine Plasmodium T-cell antigens.
  • a Plasmodium T-cell string polypeptide construct described herein includes ten Plasmodium T-cell antigens. In some embodiments, a Plasmodium T-cell string polypeptide construct described herein comprises one or more immunogenic portions of the Plasmodium T-cell antigens. [0405] In some embodiments, a Plasmodium T-cell string polypeptide construct described herein includes a CSP polypeptide, e.g., Plasmodium CSP, e.g., P. falciparum CSP, preferably from Plasmodium falciparum isolate 3D7.
  • CSP polypeptide e.g., Plasmodium CSP, e.g., P. falciparum CSP, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes a CSP polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 1.
  • a Plasmodium T-cell string polypeptide construct described herein includes a CSP polypeptide comprising or consisting of an amino acid sequence having at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 1.
  • a Plasmodium T-cell string polypeptide construct described herein includes an antigenic Plasmodium CSP polypeptide fragment.
  • an antigenic Plasmodium CSP polypeptide fragment comprises a Plasmodium CSP N-terminal region.
  • an antigenic Plasmodium CSP polypeptide fragment comprises a Plasmodium CSP N-terminal domain.
  • an antigenic Plasmodium CSP polypeptide fragment comprises a Plasmodium CSP N-terminal region and junction region.
  • an antigenic Plasmodium CSP polypeptide fragment comprises a Plasmodium CSP N-terminal end region and junction region.
  • an antigenic Plasmodium CSP polypeptide fragment comprises a Plasmodium CSP N-terminal domain and junction region. In some embodiments, an antigenic Plasmodium CSP polypeptide fragment containing at least some part of the N-terminal domain does not contain a C-terminal region. In some embodiments, an antigenic Plasmodium CSP polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 437.
  • an antigenic Plasmodium CSP polypeptide fragment comprises or consists of the amino acid according to SEQ ID NO: 437. In some embodiments, an antigenic Plasmodium CSP polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to AILSVSSFL (SEQ ID NO: 441), LSVSSFLF (SEQ ID NO: 442), FVEALFQEY (SEQ ID NO: 443), GSSSNTRVL (SEQ ID NO: 444), or ELNYDNAGTNLY (SEQ ID NO: 445).
  • AILSVSSFL SEQ ID NO: 441
  • LSVSSFLF SEQ ID NO: 442
  • FVEALFQEY SEQ ID NO: 443
  • GSSSNTRVL SEQ ID NO: 444
  • an antigenic Plasmodium CSP polypeptide fragment comprises or consists of the amino acid sequence AILSVSSFL (SEQ ID NO: 441), LSVSSFLF (SEQ ID NO: 442), FVEALFQEY (SEQ ID NO: 443), GSSSNTRVL (SEQ ID NO: 444), and/or ELNYDNAGTNLY (SEQ ID NO: 445).
  • a Plasmodium T-cell string polypeptide construct described herein includes a LSA-1(a) polypeptide, e.g., Plasmodium LSA-1(a) polypeptide.
  • a Plasmodium T-cell string polypeptide construct described herein includes a LSA-1(a) polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 424. In some embodiments, a Plasmodium T-cell string polypeptide construct described herein includes a LSA-1(a) polypeptide comprising or consisting of an amino acid sequence having at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 424.
  • a Plasmodium T-cell string polypeptide construct described herein includes an antigenic Plasmodium LSA-1(a) polypeptide fragment.
  • an antigenic Plasmodium LSA-1(a) polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 447.
  • an antigenic Plasmodium LSA-1(a) polypeptide fragment comprises or consists of the amino acid according to SEQ ID NO: 447.
  • an antigenic Plasmodium LSA-1(a) polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to HGDVLAEDLY (SEQ ID NO: 451), VLAEDLYGRL (SEQ ID NO: 452), KSADIQNHTL (SEQ ID NO: 453), IQNHTLETV (SEQ ID NO: 454), and/or ISDVNDFQISKY (SEQ ID NO: 455).
  • an antigenic Plasmodium LSA-1(a) polypeptide fragment comprises or consists of the amino acid sequence HGDVLAEDLY (SEQ ID NO: 451), VLAEDLYGRL (SEQ ID NO: 452), KSADIQNHTL (SEQ ID NO: 453), IQNHTLETV (SEQ ID NO: 454), and/or ISDVNDFQISKY (SEQ ID NO: 455).
  • a Plasmodium T-cell string polypeptide construct described herein includes a LSA-1(b) polypeptide, e.g., Plasmodium LSA-1(b) polypeptide.
  • a Plasmodium T-cell string polypeptide construct described herein includes a LSA-1(b) polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 425.
  • a Plasmodium T-cell string polypeptide construct described herein includes a LSA-1(b) polypeptide comprising or consisting of an amino acid sequence having at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 425.
  • a Plasmodium T-cell string polypeptide construct described herein includes an antigenic Plasmodium LSA-1(b) polypeptide fragment.
  • an antigenic Plasmodium LSA-1(b) polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 457.
  • an antigenic Plasmodium LSA-1(b) polypeptide fragment comprises or consists of the amino acid according to SEQ ID NO: 457.
  • an antigenic Plasmodium LSA-1(b) polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to DEFKPIVQY (SEQ ID NO: 461), FQDEENIGI (SEQ ID NO: 462), FQDEENIGIY (SEQ ID NO: 463), KSLYDEHIKKY (SEQ ID NO: 464), SLYDEHIKK (SEQ ID NO: 465), SLYDEHIKKYK (SEQ ID NO: 466), YDEHIKKY (SEQ ID NO: 467), HIFDGDNEILQI (SEQ ID NO: 468), IVDELSEDITKY (SEQ ID NO: 469), and/or SEDITKYFM (SEQ ID NO: 470).
  • DEFKPIVQY SEQ ID NO
  • an antigenic Plasmodium LSA-1(b) polypeptide fragment comprises or consists of the amino acid sequence DEFKPIVQY (SEQ ID NO: 461), FQDEENIGI (SEQ ID NO: 462), FQDEENIGIY (SEQ ID NO: 463), KSLYDEHIKKY (SEQ ID NO: 464), SLYDEHIKK (SEQ ID NO: 465), SLYDEHIKKYK (SEQ ID NO: 466), YDEHIKKY (SEQ ID NO: 467), HIFDGDNEILQI (SEQ ID NO: 468), IVDELSEDITKY (SEQ ID NO: 469), and/or SEDITKYFM (SEQ ID NO: 470).
  • a Plasmodium T-cell string polypeptide construct described herein includes a TRAP polypeptide, e.g., Plasmodium TRAP polypeptide.
  • a Plasmodium T-cell string polypeptide construct described herein includes a TRAP polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 422.
  • a Plasmodium T-cell string polypeptide construct described herein includes a TRAP polypeptide comprising or consisting of an amino acid sequence having at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 422.
  • a Plasmodium T-cell string polypeptide construct described herein includes an antigenic Plasmodium TRAP polypeptide fragment.
  • an antigenic Plasmodium TRAP polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 472.
  • an antigenic Plasmodium TRAP polypeptide fragment comprises or consists of the amino acid according to SEQ ID NO: 472.
  • an antigenic Plasmodium TRAP polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to LLMDCSGSI (SEQ ID NO: 476), GSIRRHNW (SEQ ID NO: 477), NLNDNAIHL (SEQ ID NO: 478), NLNDNAIHLY (SEQ ID NO: 479), LNDNAIHLY (SEQ ID NO: 480), ALIIIKSLL (SEQ ID NO: 481), SLLSTNLPY (SEQ ID NO: 482), SLLSTNLPYGK (SEQ ID NO: 483), NLTDALLQV (SEQ ID NO: 484), HLNDRINRENANQ (SEQ ID NO: 485), KLSDRGVKI (SEQ ID NO: 486), K
  • an antigenic Plasmodium TRAP polypeptide fragment comprises or consists of the amino acid sequence to LLMDCSGSI (SEQ ID NO: 476), GSIRRHNW (SEQ ID NO: 477), NLNDNAIHL (SEQ ID NO: 478), NLNDNAIHLY (SEQ ID NO: 479), LNDNAIHLY (SEQ ID NO: 480), ALIIIKSLL (SEQ ID NO: 481), SLLSTNLPY (SEQ ID NO: 482), SLLSTNLPYGK (SEQ ID NO: 483), NLTDALLQV (SEQ ID NO: 484), HLNDRINRENANQ (SEQ ID NO: 485), KLSDRGVKI (SEQ ID NO: 486), KLSDRGVKIAV (SEQ ID NO: 487), AVFGIGQGINV (SEQ ID NO: 488), AVFGIGQGINVA (SEQ ID NO: 489), and/or FGIGQGINV (SEQ ID NO:
  • a Plasmodium T-cell string polypeptide construct described herein includes a LSAP1 polypeptide, e.g., Plasmodium LSAP1 polypeptide.
  • a Plasmodium T-cell string polypeptide construct described herein includes a LSAP1 polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 427.
  • a Plasmodium T-cell string polypeptide construct described herein includes a LSAP1 polypeptide comprising or consisting of an amino acid sequence having at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 427.
  • a Plasmodium T-cell string polypeptide construct described herein includes an antigenic Plasmodium LSAP1 polypeptide fragment.
  • an antigenic Plasmodium LSAP1 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 492.
  • an antigenic Plasmodium LSAP1 polypeptide fragment comprises or consists of the amino acid according to SEQ ID NO: 492.
  • a Plasmodium T-cell string polypeptide construct described herein includes a LSAP2 polypeptide, e.g., Plasmodium LSAP2 polypeptide.
  • a Plasmodium T-cell string polypeptide construct described herein includes a LSAP2 polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 428. In some embodiments, a Plasmodium T-cell string polypeptide construct described herein includes a LSAP2 polypeptide comprising or consisting of an amino acid sequence having at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 428.
  • a Plasmodium T-cell string polypeptide construct described herein includes an antigenic Plasmodium LSAP2 polypeptide fragment.
  • an antigenic Plasmodium LSAP2 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 497.
  • an antigenic Plasmodium LSAP2 polypeptide fragment comprises or consists of the amino acid according to SEQ ID NO: 497.
  • an antigenic Plasmodium LSAP2 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to GLKPSDLNRK (SEQ ID NO: 501), VTEEDLERM (SEQ ID NO: 502), LIWHYSHSL (SEQ ID NO: 503), SLWSICGKL (SEQ ID NO: 504), LAHEHKLPF (SEQ ID NO: 505), HVTDELLIK (SEQ ID NO: 506), HDDYNSIY (SEQ ID NO: 507), and/or HDDYNSIYNY (SEQ ID NO: 508).
  • GLKPSDLNRK SEQ ID NO: 501
  • VTEEDLERM SEQ ID NO: 502
  • LIWHYSHSL SEQ ID NO: 503
  • SLWSICGKL
  • an antigenic Plasmodium LSAP2 polypeptide fragment comprises or consists of the amino acid sequence GLKPSDLNRK (SEQ ID NO: 501), VTEEDLERM (SEQ ID NO: 502), LIWHYSHSL (SEQ ID NO: 503), SLWSICGKL (SEQ ID NO: 504), LAHEHKLPF (SEQ ID NO: 505), HVTDELLIK (SEQ ID NO: 506), HDDYNSIY (SEQ ID NO: 507), and/or HDDYNSIYNY (SEQ ID NO: 508).
  • GLKPSDLNRK SEQ ID NO: 501
  • VTEEDLERM SEQ ID NO: 502
  • LIWHYSHSL SEQ ID NO: 503
  • SLWSICGKL SEQ ID NO: 504
  • LAHEHKLPF SEQ ID NO: 505
  • HVTDELLIK SEQ ID NO: 506
  • HDDYNSIY SEQ ID NO: 507
  • HDDYNSIYNY
  • a Plasmodium T-cell string polypeptide construct described herein includes a UIS3 polypeptide, e.g., Plasmodium UIS3 polypeptide.
  • a Plasmodium T-cell string polypeptide construct described herein includes a UIS3 polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 434.
  • a Plasmodium T-cell string polypeptide construct described herein includes a UIS3 polypeptide comprising or consisting of an amino acid sequence having at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 434.
  • a Plasmodium T-cell string polypeptide construct described herein includes an antigenic Plasmodium UIS3 polypeptide fragment.
  • an antigenic Plasmodium UIS3 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 510.
  • an antigenic Plasmodium UIS3 polypeptide fragment comprises or consists of the amino acid according to SEQ ID NO: 410.
  • an antigenic Plasmodium UIS3 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SLIASGAIASV (SEQ ID NO: 514).
  • an antigenic Plasmodium UIS3 polypeptide fragment comprises or consists of the amino acid sequence SLIASGAIASV (SEQ ID NO: 514).
  • a Plasmodium T-cell string polypeptide construct described herein includes an ETRAMP10.3 polypeptide, e.g., Plasmodium ETRAMP10.3 polypeptide.
  • a Plasmodium T-cell string polypeptide construct described herein includes an ETRAMP10.3 polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 435.
  • a Plasmodium T-cell string polypeptide construct described herein includes an ETRAMP10.3 polypeptide comprising or consisting of an amino acid sequence having at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 435.
  • a Plasmodium T- cell string polypeptide construct described herein includes an antigenic Plasmodium ETRAMP10.3 polypeptide fragment.
  • an antigenic Plasmodium ETRAMP10.3 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 516.
  • an antigenic Plasmodium ETRAMP10.3 polypeptide fragment comprises or consists of the amino acid according to SEQ ID NO: 516.
  • an antigenic Plasmodium ETRAMP10.3 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SLLGCVLTL (SEQ ID NO: 520), RTLEKLLRK (SEQ ID NO: 521), RTLEKLLRKK (SEQ ID NO: 522), and/or GLFGSLGYK (SEQ ID NO: 523).
  • an antigenic Plasmodium ETRAMP10.3 polypeptide fragment comprises or consists of the amino acid sequence SLLGCVLTL (SEQ ID NO: 520), RTLEKLLRK (SEQ ID NO: 521), RTLEKLLRKK (SEQ ID NO: 522), and/or GLFGSLGYK (SEQ ID NO: 523).
  • a Plasmodium T-cell string polypeptide construct described herein includes a LISP-1 polypeptide, e.g., Plasmodium LISP-1 polypeptide.
  • a Plasmodium T-cell string polypeptide construct described herein includes a LISP-1 polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 429. In some embodiments, a Plasmodium T-cell string polypeptide construct described herein includes a LISP-1 polypeptide comprising or consisting of an amino acid sequence having at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 429.
  • a Plasmodium T-cell string polypeptide construct described herein includes an antigenic Plasmodium LISP-1 polypeptide fragment.
  • an antigenic Plasmodium LISP-1 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 525.
  • an antigenic Plasmodium LISP-1 polypeptide fragment comprises or consists of the amino acid according to SEQ ID NO: 525.
  • an antigenic Plasmodium LISP-1 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to YTVGDVLRY (SEQ ID NO: 529), SIYIFHEKK (SEQ ID NO: 530), and/or KIFGCITNK (SEQ ID NO: 531).
  • an antigenic Plasmodium LISP-1 polypeptide fragment comprises or consists of the amino acid YTVGDVLRY (SEQ ID NO: 529), SIYIFHEKK (SEQ ID NO: 530), and/or KIFGCITNK (SEQ ID NO: 531).
  • a Plasmodium T-cell string polypeptide construct described herein includes a LISP-2 polypeptide, e.g., Plasmodium LISP-2 polypeptide.
  • a Plasmodium T-cell string polypeptide construct described herein includes a LISP-2 polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 430.
  • a Plasmodium T-cell string polypeptide construct described herein includes a LISP-2 polypeptide comprising or consisting of an amino acid sequence having at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 430.
  • a Plasmodium T ll string polypeptide construct described herein includes an antigenic Plasmodium LISP-2 polypeptide fragment.
  • an antigenic Plasmodium LISP-2 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 533.
  • an antigenic Plasmodium LISP-2 polypeptide fragment comprises or consists of the amino acid according to SEQ ID NO: 533.
  • an antigenic Plasmodium LISP-2 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to ALNIHVMSK (SEQ ID NO: 537), ALNIHVMSKY (SEQ ID NO: 538), NVENRINNISNHY (SEQ ID NO: 539), RLFFLLFYK (SEQ ID NO: 540), and/or KIYYKTKHFEK (SEQ ID NO: 541).
  • an antigenic Plasmodium LISP-2 polypeptide fragment comprises or consists of the amino acid sequence ALNIHVMSK (SEQ ID NO: 537), ALNIHVMSKY (SEQ ID NO: 538), NVENRINNISNHY (SEQ ID NO: 539), RLFFLLFYK (SEQ ID NO: 540), and/or KIYYKTKHFEK (SEQ ID NO: 541).
  • a Plasmodium T-cell string polypeptide construct described herein includes a LSA-3 polypeptide, e.g., Plasmodium LSA-3 polypeptide.
  • a Plasmodium T-cell string polypeptide construct described herein includes a LSA-3 polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 426. In some embodiments, a Plasmodium T-cell string polypeptide construct described herein includes a LSA-3 polypeptide comprising or consisting of an amino acid sequence having at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 426.
  • a Plasmodium T-cell string polypeptide construct described herein includes an antigenic Plasmodium LSA-3 polypeptide fragment.
  • an antigenic Plasmodium LSA-3 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 543.
  • an antigenic Plasmodium LSA-3 polypeptide fragment comprises or consists of the amino acid according to SEQ ID NO: 543.
  • an antigenic Plasmodium LSA-3 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to KLIEETQEL (SEQ ID NO: 547), VEADLIKDM (SEQ ID NO: 548), EEHDITTTL (SEQ ID NO: 549), TTLDEVVEL (SEQ ID NO: 550), TLDEVVEL (SEQ ID NO: 551), ELESEILEDY (SEQ ID NO: 552), LESEILEDY (SEQ ID NO: 553), KTIETDILEEK (SEQ ID NO: 554), and/or ETDILEEKK (SEQ ID NO: 555).
  • KLIEETQEL SEQ ID NO: 547
  • VEADLIKDM SEQ ID NO: 548
  • EEHDITTTL SEQ ID NO: 549
  • an antigenic Plasmodium LSA-3 polypeptide fragment comprises or consists of the amino acid sequence KLIEETQEL (SEQ ID NO: 547), VEADLIKDM (SEQ ID NO: 548), EEHDITTTL (SEQ ID NO: 549), TTLDEVVEL (SEQ ID NO: 550), TLDEVVEL (SEQ ID NO: 551), ELESEILEDY (SEQ ID NO: 552), LESEILEDY (SEQ ID NO: 553), KTIETDILEEK (SEQ ID NO: 554), and/or ETDILEEKK (SEQ ID NO: 555).
  • a Plasmodium T-cell string polypeptide construct described herein includes an EXP1 polypeptide, e.g., Plasmodium EXP1 polypeptide.
  • a Plasmodium T-cell string polypeptide construct described herein includes an EXP1 polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 431.
  • a Plasmodium T-cell string polypeptide construct described herein includes a EXP1 polypeptide comprising or consisting of an amino acid sequence having at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 431.
  • a Plasmodium T-cell string polypeptide construct described herein includes an antigenic Plasmodium EXP1 polypeptide fragment.
  • an antigenic Plasmodium EXP1 polypeptide fragment comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 557.
  • an antigenic Plasmodium EXP1 polypeptide fragment comprises or consists of the amino acid according to SEQ ID NO: 557. 3. Trafficking Signals [0417]
  • a Plasmodium T-cell string polypeptide construct described herein includes a trafficking signal.
  • a trafficking signal is an MHC class I trafficking signal (MITD).
  • the MITD comprises or consists of an amino acid sequence according to SEQ ID NO: 561. 4. Select Embodiments of Plasmodium T-cell String Peptide Constructs [0418] The present disclosure provides, among other things, polyribonucleotides that encode a Plasmodium T- cell string polypeptide construct.
  • a Plasmodium T-cell string polypeptide construct described herein includes two or more of: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium LSA-1 (e.g., LSA-1(a) and/or LSA-1(b)) polypeptide fragment; (iii) an antigenic Plasmodium TRAP polypeptide fragment; (iv) an antigenic Plasmodium LSAP2 polypeptide fragment; (v) an antigenic Plasmodium UIS3 polypeptide fragment; (vi) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (vii) an antigenic Plasmodium LISP-1 polypeptide fragment; (viii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (ix) an antigenic Plasmodium LSA-3 polypeptide fragment.
  • a Plasmodium T ll string polypeptide construct described herein includes one or more polypeptide or portions (e.g., antigenic portions) thereof from Plasmodium falciparum.
  • one or more polypeptide or portions (e.g., antigenic portions) thereof are one or more P. falciparum T cell antigens.
  • one or more P. falciparum T cell antigens are from P. falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein does not include one or more polypeptide or portions (e.g., antigenic portions) thereof from Plasmodium berghei (e.g., antigenic Plasmodium berghei CSP polypeptide fragments).
  • a Plasmodium T-cell string polypeptide construct describes herein does not include an antigenic fragment of a bacterial polypeptide.
  • a Plasmodium T-cell string polypeptide construct describes herein does not include an antigenic bacillus Calmette-Guérin (BCG) polypeptide fragment.
  • BCG antigenic bacillus Calmette-Guérin
  • an antigenic BCG polypeptide fragment comprises an amino acid sequence according to SEQ ID NO: 416.
  • a Plasmodium T-cell string polypeptide construct describes herein does not include an antigenic tetanus toxin (TT) polypeptide fragment.
  • an antigenic TT polypeptide fragment comprises an amino acid sequence according to SEQ ID NO: 417.
  • a Plasmodium T-cell string polypeptide construct describes herein does not include an antigenic Plasmodium sporozoite threonine–asparagine-rich protein (STARP) polypeptide fragment.
  • an antigenic Plasmodium STARP polypeptide fragment comprises an amino acid sequence according to SEQ ID NO: 418.
  • a Plasmodium T-cell string polypeptide construct described herein includes one or more Plasmodium polypeptide regions or portions (e.g., antigenic portions) thereof, as described above. Exemplary constructs are described below. Exemplary descriptions and sequences for the one or more Plasmodium polypeptide regions or portions (e.g., antigenic portions) thereof discussed below are further discussed in Section IIA(ii) above.
  • a Plasmodium T-cell string polypeptide construct described herein includes a CSP polypeptide (or one or more antigenic Plasmodium CSP polypeptide fragments) as described above, a Plasmodium TRAP polypeptide (or one or more antigenic Plasmodium TRAP polypeptide fragments) as described above, a Plasmodium LSA-1(a) polypeptide (or one or more antigenic Plasmodium LSA-1(a) polypeptide fragments) as described above, a Plasmodium LSA-1(b) polypeptide (or one or more antigenic Plasmodium LSA-1(b) polypeptide fragments) as described above, a Plasmodium LSA-3 polypeptide (or one or more antigenic Plasmodium LSA-3 polypeptide fragments) as described above, and a Plasmodium LSAP2
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (iv) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (v) an antigenic Plasmodium LSA-3 polypeptide fragment; and (vi) an antigenic Plasmodium LSAP2 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (iv) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (v) an antigenic Plasmodium LSA-3 polypeptide fragment; and (vi) an antigenic Plasmodium LSAP2 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO:167.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 167.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (iii) an antigenic Plasmodium LSAP2 polypeptide fragment; (iv) an antigenic Plasmodium CSP polypeptide fragment; (v) an antigenic Plasmodium LSA-3 polypeptide fragment; and (vi) an antigenic Plasmodium TRAP polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 170.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 170.
  • a Plasmodium T-cell string polypeptide construct described herein includes a Plasmodium LSAP1 polypeptide (or one or more antigenic Plasmodium LSAP1 polypeptide fragments) as described above, a Plasmodium EXP1 polypeptide (or one or more antigenic Plasmodium EXP1 polypeptide fragments) as described above, a Plasmodium UIS3 polypeptide (or one or more antigenic Plasmodium UIS3 polypeptide fragments) as described above, a Plasmodium ETRAMP10.3 polypeptide (or one or more antigenic Plasmodium ETRAMP10.3 polypeptide fragments) as described above, a Plasmodium LISP-1 polypeptide (or one or more antigenic Plasmodium LISP-1 polypeptide fragments) as described above, and a Plasmodium LISP-2 polypeptide
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium LSAP1 polypeptide fragment; (ii) an antigenic Plasmodium EXP1 polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LISP-1 polypeptide fragment; and (vi) an antigenic Plasmodium LISP-2 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium EXP1 polypeptide fragment; (ii) an antigenic Plasmodium UIS3 polypeptide fragment; (iii) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (iv) an antigenic Plasmodium LSAP1 polypeptide fragment; (v) an antigenic Plasmodium LISP-2 polypeptide fragment; and (vi) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 173.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 173.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium UIS3 polypeptide fragment; (ii) an antigenic Plasmodium LSAP1 polypeptide fragment; (iii) an antigenic Plasmodium LISP-1 polypeptide fragment; (iv) an antigenic Plasmodium EXP1 polypeptide fragment; (v) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; and (vi) an antigenic Plasmodium LISP-2 polypeptide fragment.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 176.
  • a Pl i T ll string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 176.
  • a Plasmodium T-cell string polypeptide construct described herein includes a CSP polypeptide (or one or more antigenic Plasmodium CSP polypeptide fragments) as described above, a Plasmodium TRAP polypeptide (or one or more antigenic Plasmodium TRAP polypeptide fragments) as described above, a Plasmodium LSAP2 polypeptide (or one or more antigenic Plasmodium LSAP2 polypeptide fragments) as described above, a Plasmodium UIS3 polypeptide (or one or more antigenic Plasmodium UIS3 polypeptide fragments) as described above, and a Plasmodium ETRAMP10.3 polypeptide (or one or more antigenic Plasmodium ETRAMP10.3 polypeptide fragments) as described above, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plas
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; and (v) an antigenic Plasmodium LSAP2 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; and (v) an antigenic Plasmodium LSAP2 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 179.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 179.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; and (v) an antigenic Plasmodium LSAP2 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 221.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 221.
  • a Plasmodium T-cell string polypeptide construct described herein includes a CSP polypeptide (or one or more antigenic Plasmodium CSP polypeptide fragments) as described above, a Plasmodium TRAP polypeptide (or one or more antigenic Plasmodium TRAP polypeptide fragments) as described above, a Plasmodium LSA-1(a) polypeptide (or one or more antigenic Plasmodium LSA-1(a) polypeptide fragments) as described above, a Plasmodium LSA-1(b) polypeptide (or one or more antigenic Plasmodium LSA-1(b) polypeptide fragments) as described above, a Plasmodium LSA-3 polypeptide (or one or more antigenic Plasmodium LSA-3 polypeptide fragments) as described above, a Plasmodium LSA-3 polypeptide (or one or more antigenic Plasmodium LSA-3 polypeptide fragments) as described above, a Plasmodium LSA-3 polypeptide (or
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA-3 polypeptide fragment; (vii) an antigenic Plasmodium LSA-1(a) polypeptide fragment; and (viii) an antigenic Plasmodium LSA-1(b) polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA-3 polypeptide fragment; (vii) an antigenic Plasmodium LSA-1(a) polypeptide fragment; and (viii) an antigenic Plasmodium LSA-1(b) polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 182.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 182.
  • a Plasmodium T-cell string polypeptide construct described herein includes a CSP polypeptide (or one or more antigenic Plasmodium CSP polypeptide fragments) as described above, a Plasmodium TRAP polypeptide (or one or more antigenic Plasmodium TRAP polypeptide fragments) as described above, a Plasmodium LSA-1(a) polypeptide (or one or more antigenic Plasmodium LSA-1(a) polypeptide fragments) as described above, a Plasmodium LSA-1(b) polypeptide (or one or more antigenic Plasmodium LSA-1(b) polypeptide fragments) as described above, a Plasmodium LSAP2 polypeptide (or one or more antigenic Plasmodium LSAP2 polypeptide fragments) as described
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (vii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (viii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (ix) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (vii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (viii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (ix) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 188
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 188.
  • a Plasmodium T-cell string polypeptide construct described herein includes a CSP polypeptide (or one or more antigenic Plasmodium CSP polypeptide fragments) as described above, a Plasmodium TRAP polypeptide (or one or more antigenic Plasmodium TRAP polypeptide fragments) as described above, a Plasmodium LSA-1(a) polypeptide (or one or more antigenic Plasmodium LSA-1(a) polypeptide fragments) as described above, a Plasmodium LSA-1(b) polypeptide (or one or more antigenic Plasmodium LSA-1(b) polypeptide fragments) as described above, a Plasmodium LSAP2 polypeptide (or one or more antigenic Plasmodium LSAP2 polypeptide fragments) as described above, a
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (vii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; and (viii) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (vii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; and (viii) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 191.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 191.
  • a Plasmodium T-cell string polypeptide construct described herein includes a CSP polypeptide (or one or more antigenic Plasmodium CSP polypeptide fragments) as described above, a Plasmodium TRAP polypeptide (or one or more antigenic Plasmodium TRAP polypeptide fragments) as described above, a Plasmodium LSAP2 polypeptide (or one or more antigenic Plasmodium LSAP2 polypeptide fragments) as described above, a Plasmodium UIS3 polypeptide (or one or more antigenic Plasmodium UIS3 polypeptide fragments) as described above, a Plasmodium ETRAMP10.3 polypeptide (or one or more antigenic Plasmodium ETRAMP10.3 polypeptide fragments) as described above, a Plasmodium LISP-1 polypeptide (or one or more anti
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LISP-2 polypeptide fragment; and (vii) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LISP-2 polypeptide fragment; and (vii) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 194.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 194.
  • a Plasmodium T-cell string polypeptide construct described herein includes a CSP polypeptide (or one or more antigenic Plasmodium CSP polypeptide fragments) as described above, a Plasmodium TRAP polypeptide (or one or more antigenic Plasmodium TRAP polypeptide fragments) as described above, a Plasmodium LSA-1(b) polypeptide (or one or more antigenic Plasmodium LSA-1(b) polypeptide fragments) as described above, a Plasmodium LSAP2 polypeptide (or one or more antigenic Plasmodium LSAP2 polypeptide fragments) as described above, a Plasmodium UIS3 polypeptide (or one or more antigenic Plasmodium UIS3 polypeptide fragments) as described above, a Plasmodium ETRAMP10.3 polypeptide
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA-1(b) polypeptide fragment; and (vii) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA-1(b) polypeptide fragment; and (vii) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 197.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 197.
  • a Plasmodium T-cell string polypeptide construct described herein includes a CSP polypeptide (or one or more antigenic Plasmodium CSP polypeptide fragments) as described above, a Plasmodium TRAP polypeptide (or one or more antigenic Plasmodium TRAP polypeptide fragments) as described above, a Plasmodium LSA-1(a) polypeptide (or one or more antigenic Plasmodium LSA-1(a) polypeptide fragments) as described above, a Plasmodium LSA-1(b) polypeptide (or one or more antigenic Plasmodium LSA-1(b) polypeptide fragments) as described above, a Plasmodium LSA-3 polypeptide (or one or more antigenic Plasmodium LSA-3 polypeptide fragments)
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (vii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (viii) an antigenic Plasmodium LISP-2 polypeptide fragment; (ix) an antigenic Plasmodium LISP-1 polypeptide fragment; and (x) an antigenic Plasmodium LSA-3 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium CSP polypeptide fragment; (ii) an antigenic Plasmodium TRAP polypeptide fragment; (iii) an antigenic Plasmodium UIS3 polypeptide fragment; (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (v) an antigenic Plasmodium LSAP2 polypeptide fragment; (vi) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (vii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (viii) an antigenic Plasmodium LISP-2 polypeptide fragment; (ix) an antigenic Plasmodium LISP-1 polypeptide fragment; and (x) an antigenic Plasmodium LSA-3 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 200.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 200.
  • a Plasmodium T-cell string polypeptide construct described herein includes a Plasmodium LSA-1(a) polypeptide (or one or more antigenic Plasmodium LSA-1(a) polypeptide fragments) as described above, a Plasmodium LSA-1(b) polypeptide (or one or more antigenic Plasmodium LSA-1(b) polypeptide fragments) as described above, a Plasmodium LISP-1 polypeptide (or one or more antigenic Plasmodium LISP-1 polypeptide fragments) as described above, and a Plasmodium LISP-2 polypeptide (or one or more antigenic Plasmodium LISP-2 polypeptide fragments) as described above, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (iii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (iii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 209.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 209.
  • a Plasmodium T-cell string polypeptide construct described herein includes a Plasmodium LSA-1(a) polypeptide (or one or more antigenic Plasmodium LSA-1(a) polypeptide fragments) as described above, a Plasmodium LSA-1(b) polypeptide (or one or more antigenic Plasmodium LSA-1(b) polypeptide fragments) as described above, a Plasmodium LSA-3 polypeptide (or one or more antigenic Plasmodium LSA-3 polypeptide fragments) as described above, a Plasmodium LISP-1 polypeptide (or one or more antigenic Plasmodium LISP-1 polypeptide fragments) as described above, and a Plasmodium LISP-2 polypeptide (or one or more antigenic Plasmodium LISP-2 polypeptide fragments) as described above, wherein the Plasmodium is
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (iii) an antigenic Plasmodium LISP-2 polypeptide fragment; (iv) an antigenic Plasmodium LISP-1 polypeptide fragment; and (v) an antigenic Plasmodium LSA-3 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium LSA- 1(a) polypeptide fragment; (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (iii) an antigenic Plasmodium LISP-2 polypeptide fragment; (iv) an antigenic Plasmodium LISP-1 polypeptide fragment; and (v) an antigenic Plasmodium LSA-3 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 212.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 212.
  • a Plasmodium T-cell string polypeptide construct described herein includes a Plasmodium TRAP polypeptide (or one or more antigenic Plasmodium TRAP polypeptide fragments) as described above, a Plasmodium LSA-1(a) polypeptide (or one or more antigenic Plasmodium LSA-1(a) polypeptide fragments) as described above, a Plasmodium LSA-1(b) polypeptide (or one or more antigenic Plasmodium LSA-1(b) polypeptide fragments) as described above, a Plasmodium LSAP2 polypeptide (or one or more antigenic Plasmodium LSAP2 polypeptide fragments) as described above, a Plasmodium UIS3 polypeptide (or one or more antigenic Plasmodium UIS3 polypeptide fragments)
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium TRAP polypeptide fragment; (ii) an antigenic Plasmodium UIS3 polypeptide fragment; (iii) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (iv) an antigenic Plasmodium LSAP2 polypeptide fragment; (v) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (vi) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (vii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (viii) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium TRAP polypeptide fragment; (ii) an antigenic Plasmodium UIS3 polypeptide fragment; (iii) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; (iv) an antigenic Plasmodium LSAP2 polypeptide fragment; (v) an antigenic Plasmodium LSA-1(a) polypeptide fragment; (vi) an antigenic Plasmodium LSA-1(b) polypeptide fragment; (vii) an antigenic Plasmodium LISP-2 polypeptide fragment; and (viii) an antigenic Plasmodium LISP-1 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 215.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 215.
  • a Plasmodium T-cell string polypeptide construct described herein includes a Plasmodium TRAP polypeptide (or one or more antigenic Plasmodium TRAP polypeptide fragments) as described above, a Plasmodium LSAP2 polypeptide (or one or more antigenic Plasmodium LSAP2 polypeptide fragments) as described above, a Plasmodium UIS3 polypeptide (or one or more antigenic Plasmodium UIS3 polypeptide fragments) as described above, and a Plasmodium ETRAMP10.3 polypeptide (or one or more antigenic Plasmodium ETRAMP10.3 polypeptide fragments) as described above, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes: (i) an antigenic Plasmodium TRAP polypeptide fragment; (ii) an antigenic Plasmodium UIS3 polypeptide fragment; (iii) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; and (iv) an antigenic Plasmodium LSAP2 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes in order: (i) an antigenic Plasmodium TRAP polypeptide fragment; (ii) an antigenic Plasmodium UIS3 polypeptide fragment; (iii) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment; and (iv) an antigenic Plasmodium LSAP2 polypeptide fragment, wherein the Plasmodium is preferably Plasmodium falciparum and more preferably Plasmodium falciparum isolate 3D7.
  • a Plasmodium T-cell string polypeptide construct described herein includes an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to an amino acid sequence according to SEQ ID NO: 218.
  • a Plasmodium T-cell string polypeptide construct described herein includes the amino acid sequence of SEQ ID NO: 218.
  • Plasmodium T-cell string polypeptide construct described herein has an amino acid sequence provided in Table 3, and/or is encoded by a nucleotide sequence provided in Table 9.
  • Exemplary Plasmodium T-cell string polypeptide constructs are also shown schematically in FIG.3.
  • Table 3 Amino Acid Sequences Encoding Exemplary Plasmodium T-cell String Polypeptide Constructs
  • Plasmodium CSP Polypeptide Constructs utilizes RNA technologies as a modality to express one or more Plasmodium CSP polypeptide constructs (also referred to as “Plasmodium CSP polypeptides,” “malaria CSP polypeptide constructs” or “malarial polypeptide constructs”).
  • a Plasmodium CSP polypeptide construct as described herein can include one or more Plasmodium CSP polypeptide regions or portions (e.g., antigenic portions) thereof.
  • a Plasmodium CSP polypeptide construct as described herein includes one or more polypeptides or antigenic portions thereof that are expressed during a Plasmodium sporozoite stage.
  • Plasmodium CSP polypeptide constructs as described herein is capable of eliciting antibody responses, preferably strong antibody responses. Methods to determine antibody responses are well known in the art and described in the examples.
  • Plasmodium CSP polypeptide constructs as described herein can be (i) full length Plasmodium CSP protein including the Plasmodium secretory signal and GPI anchor or (ii) only contain selected regions or portions (such as antigenic portions) of a Plasmodium CSP protein. [0453] A portion of a CSP polypeptide or region can be a characteristic portion of CSP polypeptide or region.
  • a Plasmodium CSP polypeptide construct additionally includes one or more additional amino acid sequences, such as a secretory signal (e.g., a heterologous secretory signal), a transmembrane region (e.g., a heterologous transmembrane region), a helper antigen, a multimerization region, and/or a linker, as described herein.
  • a Plasmodium CSP polypeptide construct described herein includes one or more regions or portions (e.g., antigenic portions) of a CSP, e.g., Plasmodium CSP, e.g., P.
  • a region of CSP may refer to a N-terminal region, a N- terminal end region, a junction region (or its sub-regions: an R1 region or a junction), a minor repeat region, a major repeat region or a C-terminal region.
  • a portion of CSP may refer to parts of a CSP polypeptide region or parts spanning two or more CSP polypeptide regions.
  • a portion of CSP may refer to an N-terminal domain, a central domain, or a C-terminal domain.
  • a CSP polypeptide portion (e.g., antigenic portions) comprises 25, 30, 35, 40, or 45 contiguous amino acids of the amino acid sequence according to SEQ ID NO: 1.
  • a Plasmodium CSP polypeptide construct does not include a secretory signal or a transmembrane region, e.g., corresponds to amino acids 19-375 of the amino acid sequence according to SEQ ID NO: 1 or corresponds to amino acids 19-376 or 19-377 of the amino acid sequence according to SEQ ID NO: 1 (i.e., includes a serine or a serine and valine immediately after the C-terminal region).
  • a Plasmodium CSP polypeptide construct described herein includes a CSP minor repeat region.
  • a Plasmodium CSP polypeptide construct described herein includes a portion (e.g., an antigenic portion) of a CSP minor repeat region. In some embodiments, a portion (e.g., an antigenic portion) of a CSP minor repeat region is about 10, 15, 20, 21, 22, or 23 contiguous amino acids in length. In some embodiments, a Plasmodium CSP polypeptide construct described herein includes a CSP major repeat region. In some embodiments, a Plasmodium CSP polypeptide construct described herein includes a portion (e.g., an antigenic portion) of a CSP major repeat region.
  • a portion (e.g., an antigenic portion) of a CSP major repeat region is about 100, 110, 120, 130, 135, 140, 141, or 142 amino acids in length.
  • a Plasmodium CSP polypeptide construct described herein includes a CSP C-terminal region.
  • a Plasmodium CSP polypeptide construct described herein includes a portion (e.g., an antigenic portion) of a CSP C- terminal region.
  • a portion (e.g., an antigenic portion) of a CSP C-terminal region is about 80, 90, 95, 100, 101, or 102 amino acids in length.
  • a Plasmodium CSP polypeptide construct described herein includes a CSP N-terminal region. In some embodiments, a Plasmodium CSP polypeptide construct described herein includes a portion (e.g., an antigenic portion) of a CSP N-terminal region. In some embodiments, a portion (e.g., an antigenic portion) of a CSP N-terminal region is about 45, 50, 55, 60, or 61 amino acids in length. In some embodiments, a Plasmodium CSP polypeptide construct described herein includes a CSP N-terminal end region.
  • a Plasmodium CSP polypeptide construct described herein includes a portion (e.g., an antigenic portion) of a CSP N-terminal end region. In some embodiments, a portion (e.g., an antigenic portion) of a CSP N-terminal end region is about 8, 9, 10, or 11 amino acids in length. In some embodiments, a Plasmodium CSP polypeptide construct described herein includes a CSP junction region. In some embodiments, a Plasmodium CSP polypeptide construct described herein includes a portion (e.g., an antigenic portion) of a CSP junction region.
  • a portion (e.g., an antigenic portion) of a CSP junction region is about 8, 9, 10, or 11 amino acids in length.
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP minor repeat regions or portions (e.g., antigenic portions) thereof comprising one or more repeats of an amino acid sequence of NANPNVDP (SEQ ID NO: 223), and wherein the polypeptide does not comprise an amino acid sequence of NPNA (SEQ ID NO: 228), NPNANP (SEQ ID NO: 231) or NANPNA (SEQ ID NO: 232).
  • Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP polypeptide regions or portions (e.g., antigenic portions) thereof comprising one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12) repeats of an amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP polypeptide regions or portions (e.g., antigenic portions) thereof comprising two or more (e.g., between 2 and 12, or between 2 and 10, or between 2 and 9, or between 2 and 8, or between 4 and 12, or between 4 and 10) repeats of an amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP polypeptide regions or portions (e.g., antigenic portions) thereof comprising exactly three repeats of an amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP polypeptide regions or portions (e.g., antigenic portions) thereof comprising exactly eight repeats of an amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP polypeptide regions or portions (e.g., antigenic portions) thereof comprising exactly nine repeats of an amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • the repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223) are all contiguous with each other.
  • the repeats of the amino acid sequence of NANPNVDP are not all contiguous with each other.
  • a Plasmodium CSP polypeptide construct described herein comprises four portions (e.g., antigenic portions) of a Plasmodium CSP minor repeat region, and wherein each portion (e.g., antigenic portion) of a Plasmodium CSP polypeptide comprises two contiguous repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • a Plasmodium CSP minor repeat region comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 95%, or 100% identical to the amino acid sequence of SEQ ID NO: 226 (NANPNVDPNANPNVDPNANPNVDP).
  • a Plasmodium CSP minor repeat region comprises or consists of an amino acid sequence according to SEQ ID NO: 226.
  • a Plasmodium CSP polypeptide construct described herein does not comprise one or more portions of one or more Plasmodium CSP minor repeat regions (i.e., lacks or excludes a Plasmodium CSP minor repeat region or any portion thereof).
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP C-terminal regions (e.g., amino acids 273-375 of SEQ ID NO: 1), one or more Plasmodium CSP C- terminal region variants, or one or more portions (e.g., antigenic portions) thereof, wherein the C-terminal region does not include a transmembrane region.
  • Plasmodium CSP C-terminal regions e.g., amino acids 273-375 of SEQ ID NO: 1
  • portions e.g., antigenic portions
  • a Plasmodium CSP polypeptide construct described herein includes exactly one Plasmodium CSP C-terminal region, and wherein the Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to amino acids 273-375 of SEQ ID NO: 1.
  • a malarial polypeptide construct described herein includes one or more Plasmodium CSP C- terminal region variants.
  • a Plasmodium CSP C-terminal region variant comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to amino acid sequence of SEQ ID NO: 994.
  • a Plasmodium CSP polypeptide construct described herein includes exactly one portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region, and wherein the portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to amino acid sequence of SEQ ID NO: 241 (PSDKHIKEYLNKIQNSLSTEWSPCSVTCGNGIQVRIKPGSANKPKDELDYANDIEKKICKMEK).
  • a Plasmodium CSP polypeptide construct described herein includes two or more portions (e.g., antigenic portions) of a Plasmodium CSP C-terminal region (e.g., amino acids 273-375 of SEQ ID NO: 1).
  • a Plasmodium CSP polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of the Plasmodium CSP C-terminal region, wherein each of the one or more portions (e.g., antigenic portions) comprises or consists of: (i) amino acids 314-327 of SEQ ID NO:1 (or amino acids 314-327 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions); (ii) amino acids 352-363 of SEQ ID NO: 1 (or amino acids 352-363 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions); (iii) amino acids 326-374 of SEQ ID NO: 1 (or amino acids 326-374 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions), (iv) amino acids 364-377 of SEQ ID NO: 1 (or amino acids 364-377 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions), or (v) amino acids 364-377 of
  • a Plasmodium CSP polypeptide construct described herein includes one portion (e.g., antigenic portion) of the Plasmodium CSP C-terminal region, wherein the portion (e.g., antigenic portion) comprises or consists of: (i) amino acids 314-327 of SEQ ID NO: 1 (or amino acids 314-327 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions); (ii) amino acids 352-363 of SEQ ID NO: 1 (or amino acids 352-363 of SEQ ID NO:1 having 1, 2, 3, 4, or 5 amino acid substitutions); (iii) amino acids 326-374 of SEQ ID NO:1 (or amino acids 326-374 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions), (iv) amino acids 364-377 of SEQ ID NO: 1 (or amino acids 364-377 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions), or (v) a combination thereof.
  • a Plasmodium CSP polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of the Plasmodium CSP C-terminal region, wherein the one or more portions (e.g., antigenic portions) collectively comprise or consist of: (i) amino acids 314-327 of SEQ ID NO: 1 (or amino acids 314- 327 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions); (ii) amino acids 352-363 of SEQ ID NO: 1 (or amino acids 352-363 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions); (iii) amino acids 326-374 of SEQ ID NO: 1 (or amino acids 326-374 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions), (iv) amino acids 364-377 of SEQ ID NO: 1 (or amino acids 364-377 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions), or (v) amino acids 364-377 of
  • a Plasmodium CSP polypeptide construct described herein comprises a serine amino acid residue immediately following a Plasmodium CSP C-terminal region described herein. In some embodiments, a Plasmodium CSP polypeptide construct described herein comprises a serine-valine amino acid sequence immediately following a Plasmodium CSP C-terminal region described herein.
  • a Plasmodium CSP polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of the Plasmodium CSP C-terminal region, wherein the one or more portions (e.g., antigenic portions) comprise or consist of a portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region, wherein the portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region comprises an amino acid sequence according to SEQ ID NO: 261 (YLX 3 X 4 IQX 5 SLST), wherein X 3 is N or K, X 4 is K, I, or R, and X 5 is N or Y.
  • SEQ ID NO: 261 YLX 3 X 4 IQX 5 SLST
  • a Plasmodium CSP polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of the Plasmodium CSP C-terminal region, wherein the one or more portions (e.g., antigenic portions) comprise or consist of a portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region, wherein the portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region comprises an amino acid sequence according to SEQ ID NO: 262 (YLX 3 X 4 IQX 5 SLSTEW), wherein X 3 is N or K, X 4 is K, I, or R, and X 5 is N or Y.
  • SEQ ID NO: 262 YLX 3 X 4 IQX 5 SLSTEW
  • a Plasmodium CSP polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of the Plasmodium CSP C-terminal region, wherein the one or more portions (e.g., antigenic portions) comprise or consist of a portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region, wherein the portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region comprises an amino acid sequence according to SEQ ID NO: 263 (YLX 3 X 4 IQX 5 SLSTEWS), wherein X 3 is N or K, X 4 is K, I, or R, and X 5 is N or Y.
  • SEQ ID NO: 263 YLX 3 X 4 IQX 5 SLSTEWS
  • a Plasmodium CSP polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of the Plasmodium CSP C-terminal region, wherein the one or more portions (e.g., antigenic portions) comprise or consist of a portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region, wherein the portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region comprises an amino acid sequence according to SEQ ID NO: 264 (IX1X2YLX3X4IQX5SLST), wherein X1X2 is EK or KE, X3 is N or K, X4 is K, I, or R, and X5 is N or Y.
  • SEQ ID NO: 264 IX1X2YLX3X4IQX5SLST
  • a Plasmodium CSP polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of the Plasmodium CSP C-terminal region, wherein the one or more portions (e.g., antigenic portions) comprise or consist of a portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region, wherein the portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region comprises an amino acid sequence according to SEQ ID NO: 265 (IX 1 X 2 YLX 3 X 4 IQX 5 SLSTEW), wherein X 1 X 2 is EK or KE, X 3 is N or K, X 4 is K, I, or R, and X 5 is N or Y.
  • SEQ ID NO: 265 IX 1 X 2 YLX 3 X 4 IQX 5 SLSTEW
  • a Plasmodium CSP polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of the Plasmodium CSP C-terminal region, wherein the one or more portions comprise or consist of a portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region, wherein the portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region comprises an amino acid sequence according to SEQ ID NO: 266 (IX 1 X 2 YLX 3 X 4 IQX 5 SLSTEWS), wherein X 1 X 2 is EK or KE, X 3 is N or K, X 4 is K, I, or R, and X 5 is N or Y.
  • SEQ ID NO: 266 IX 1 X 2 YLX 3 X 4 IQX 5 SLSTEWS
  • a Plasmodium CSP polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of a Plasmodium CSP C-terminal region, wherein the one or more portions (e.g., antigenic portions) comprise or consist of a portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region, wherein the portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region comprises an amino acid sequence according to SEQ ID NO: 267 (IKEYLNKIQNSLSTEWS).
  • a Plasmodium CSP polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of the Plasmodium CSP C-terminal region, wherein the one or more portions (e.g., antigenic portions) comprise or consist of a portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region, wherein the portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 98%, or 100% identical to the amino acid sequence of SEQ ID NO: 241 (PSDKHIKEYLNKIQNSLSTEWSPCSVTCGNGIQVRIKPGSANKPKDELDYANDIEKKICKMEK).
  • one or more Plasmodium CSP polypeptide regions or antigenic portions thereof comprise a Plasmodium CSP C-terminal region variant or antigenic portion thereof.
  • a Plasmodium CSP C-terminal region variant or antigenic portion thereof comprises one or more amino acid substitutions, insertions, or deletions.
  • a Plasmodium CSP C-terminal region variant or antigenic portion thereof comprises one or more amino acid substitutions.
  • one or more amino acid substitutions comprise S301N, K317E, E318Q, N321K, E357Q, A361E, or any combination thereof, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • one or more amino acid substitutions comprise S301N, K317E, E318Q, N321K, E357Q, and A361E, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • a Plasmodium CSP C-terminal region variant comprises or consists of an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 994.
  • a malarial polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of a Plasmodium CSP C-terminal region variant, wherein the one or more portions comprise or consist of a portion (e.g., antigenic portion) of a Plasmodium CSP C-terminal region variant.
  • a portion of a Plasmodium CSP C-terminal region comprises an amino acid sequence according to SEQ ID NO: 992.
  • a portion of a Plasmodium CSP C-terminal region comprises an amino acid sequence according to SEQ ID NO: 993.
  • a Plasmodium CSP C-terminal region variant comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to amino acid sequence of SEQ ID NO: 994.
  • a Plasmodium CSP polypeptide construct described herein comprises a serine amino acid residue immediately following a Plasmodium CSP C-terminal region, Plasmodium CSP C-terminal region variant, or one or more portions (e.g., antigenic portions) thereof, as described herein.
  • a Plasmodium CSP polypeptide construct described herein comprises a serine-valine amino acid sequence immediately following a Plasmodium CSP C-terminal region, Plasmodium CSP C-terminal region variant, or one or more portions (e.g., antigenic portions) thereof, as described herein. [0470] In some embodiments, a Plasmodium CSP polypeptide construct described herein does not comprise one or more portions of one or more Plasmodium CSP C-terminal regions (i.e., lacks or excludes a Plasmodium CSP C-terminal region or any portion thereof).
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP junction regions or portions (e.g., antigenic portions) thereof.
  • a junction region includes an R1 region (amino acids 93-97) and a junction (SEQ ID NO: 277) at positions 98-104.
  • a Plasmodium CSP polypeptide construct described herein includes two or more Plasmodium CSP junction regions or portions (e.g., antigenic portions) thereof.
  • a Plasmodium CSP polypeptide construct described herein includes exactly one Plasmodium CSP junction region.
  • a Plasmodium CSP junction region comprises or consists of amino acids 93-104 of SEQ ID NO: 1 (or amino acids 93-104 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions). In some embodiments, a Plasmodium CSP junction region comprises or consists of an amino acid sequence that is at least 90% or at least 100% identical to the amino acid sequence of SEQ ID NO: 277. In some embodiments, a Plasmodium CSP polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of a Plasmodium CSP junction region.
  • a portion (e.g., antigenic portion) of a Plasmodium CSP junction region comprises or consists of amino acids 93-97 of SEQ ID NO: 1. In some embodiments, a portion (e.g., an antigenic portion) of a Plasmodium CSP junction region comprises or consists of amino acids 98-104 of SEQ ID NO: 1.
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP junction regions or portions (e.g., antigenic portions) thereof, wherein the Plasmodium CSP junction region comprises or consists of an amino acid sequence that is at least 90% or 100% identical to the amino acid sequence of SEQ ID NO: 272 (KLKQPADGNPDP).
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP junction regions or portions (e.g., antigenic portions) thereof, wherein the Plasmodium CSP junction region comprises or consists of an amino acid sequence according to SEQ ID NO: 272.
  • a Plasmodium CSP polypeptide construct described herein includes one or more portions (e.g., antigenic portions) of a Plasmodium CSP junction region.
  • one or more portions (e.g., antigenic portions) of a Plasmodium CSP junction region comprise a deletion of one or more of K93, L94, K95, Q96 and P97, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • one or more portions (e.g., antigenic portions) of a Plasmodium CSP junction region comprise a deletion of K93, L94, K95, and Q96, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • one or more portions (e.g., antigenic portions) of a Plasmodium CSP junction region comprise a deletion of K93, L94, K95, Q96 and P97, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP junction region variants.
  • a Plasmodium CSP junction region variant comprises one or more amino acid substitution mutations.
  • one or more substitution mutations comprise a K93A mutation, an L94A mutation, or both, wherein the amino acid numbering is relative to SEQ ID NO: 1.
  • a Plasmodium CSP junction region variant comprises the amino acid sequence of AAKQ (SEQ ID NO: 283).
  • a Plasmodium CSP polypeptide construct described herein does not comprise one or more portions of one or more Plasmodium CSP junction regions (i.e., lacks or excludes a Plasmodium CSP junction region or any portion thereof).
  • N-terminal End Region [0478] In some embodiments, a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP N-terminal end regions or portions (e.g., antigenic portions) thereof.
  • a Plasmodium CSP polypeptide construct described herein includes two or more Plasmodium CSP N-terminal end regions or portions (e.g., antigenic portions) thereof.
  • a Plasmodium CSP N-terminal end region comprises or consists of amino acids 81-92 of SEQ ID NO: 1 (or amino acids 81-92 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions).
  • a Plasmodium CSP N-terminal end region comprises or consists of an amino acid sequence that is at least 90% or at least 100% identical to the amino acid sequence of SEQ ID NO: 285 (EDNEKLRKPKHK).
  • a Plasmodium CSP N-terminal end region comprises or consists of an amino acid sequence according to SEQ ID NO: 285.
  • a Plasmodium CSP polypeptide construct described herein does not comprise a Plasmodium CSP N-terminal end region or any portion thereof (i.e., lacks or excludes a Plasmodium CSP N-terminal end region or any portion thereof).
  • N-terminal Region [0481]
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP N-terminal regions or portions (e.g., antigenic portions) thereof.
  • a Plasmodium CSP polypeptide construct described herein includes two or more Plasmodium CSP N-terminal regions or portions (e.g., antigenic portions) thereof.
  • a Plasmodium CSP N-terminal region comprises or consists of amino acids 19-80 of SEQ ID NO: 1.
  • a Plasmodium CSP N-terminal region comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100%) sequence identity to amino acids 19-80 of SEQ ID NO: 1.
  • a malarial polypeptide construct described herein includes one or more Plasmodium CSP polypeptide regions or antigenic portions thereof comprise an antigenic portion of a Plasmodium CSP N-terminal region.
  • an antigenic portion of a Plasmodium CSP N-terminal region comprises or consists of an N-terminal start region. In some embodiments, an antigenic portion of a Plasmodium CSP N-terminal region comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to the amino acid sequence of SEQ ID NO: 1010.
  • a Plasmodium CSP polypeptide construct described herein does not comprise a Plasmodium CSP N-terminal region or any portion thereof (i.e., lacks or excludes a Plasmodium CSP N-terminal region or any portion thereof).
  • Major Repeat Region [0485]
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP major repeat regions or portions (e.g., antigenic portions) thereof.
  • a Plasmodium CSP polypeptide construct described herein includes exactly one Plasmodium CSP major repeat region or portion (e.g., antigenic portion) thereof, and the Plasmodium CSP major repeat region or portion (e.g., antigenic portion) thereof comprises a total of at least 2 and at most 35 repeats of the amino acid sequence NANP (SEQ ID NO: 230).
  • a Plasmodium CSP major repeat region or portion (e.g., antigenic portion) thereof comprises two contiguous stretches of repeats of the amino acid sequence NANP (SEQ ID NO: 230), and wherein the two contiguous stretches of the repeats of the amino acid sequence NANP (SEQ ID NO: 230) flank an amino acid sequence of NVDP (SEQ ID NO: 229).
  • a Plasmodium CSP major repeat region comprises, in N-terminus to C-terminus order, 17 repeats of the amino acid sequence NANP (SEQ ID NO: 230), an amino acid sequence of NVDP (SEQ ID NO: 229), and 18 repeats of the amino acid sequence NANP (SEQ ID NO: 230).
  • a portion (e.g., antigenic portion) of the Plasmodium CSP major repeat region consists of at most 18 contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230). In some embodiments, a portion (e.g., antigenic portion) of the Plasmodium CSP major repeat region consists of 2 contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230).
  • the one or more Plasmodium CSP major repeat region or portion (e.g., antigenic portion) thereof always contains at least one repeat of the amino acid sequence of NPNANP (SEQ ID NO: 231) or NANPNA (SEQ ID NO: 232).
  • a Plasmodium CSP major repeat region comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%) sequence identity to amino acids 129-272 of SEQ ID NO: 1.
  • a Plasmodium CSP polypeptide construct described herein does not comprise a Plasmodium CSP major repeat region or a portion (e.g., antigenic portion) of a Plasmodium CSP major repeat region comprising the amino acid sequence NPNA (SEQ ID NO: 228) (i.e., lacks or excludes a Plasmodium CSP major repeat region or a portion (e.g., antigenic portion) of a Plasmodium CSP major repeat region comprising the amino acid sequence NPNA (SEQ ID NO: 228)).
  • a portion (e.g., antigenic portion) of the Plasmodium CSP major repeat region consists of at most 18 contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230). In some embodiments, a portion (e.g., antigenic portion) of the Plasmodium CSP major repeat region consists of 18 contiguous repeats of the amino acid sequence NANP (SEQ ID NO: 230). The one or more Plasmodium CSP major repeat region or portion (e.g., antigenic portion) thereof always contains at least one repeat (e.g., one instance) of the amino acid sequence of NANP (SEQ ID NO: 230).
  • a portion (e.g., antigenic portion) of the Plasmodium CSP major repeat region comprises six (6) repeats of the amino acid sequence of NANP (SEQ ID NO: 230).
  • a portion (e.g., antigenic portion) of the Plasmodium CSP major repeat region comprises or consists of an asparagine-alanine positioned immediately following the six repeats of the amino acid sequence of NANP.
  • a portion (e.g., antigenic portion) of the Plasmodium CSP major repeat region comprises or consists of the amino acid sequence SEQ ID NO: 303.
  • a Plasmodium CSP major repeat region comprises or consists of an amino acid sequence with at least 85% (e.g., at least 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%) sequence identity to SEQ ID NO: 303.
  • a Plasmodium CSP polypeptide construct described herein does not comprise a Plasmodium CSP major repeat region or a portion of a Plasmodium CSP major repeat region comprising the amino acid sequence NPNA (SEQ ID NO: 228) (i.e., lacks or excludes a Plasmodium CSP major repeat region or a portion of a Plasmodium CSP major repeat region comprising the amino acid sequence NPNA (SEQ ID NO: 228).
  • a Plasmodium CSP polypeptide construct described herein does not comprise a Plasmodium CSP major repeat region or a portion of a Plasmodium CSP major repeat region comprising the amino acid sequence NANP (SEQ ID NO: 230) (i.e., lacks or excludes a Plasmodium CSP major repeat region or a portion of a Plasmodium CSP major repeat region comprising the amino acid sequence NANP (SEQ ID NO: 230).
  • a Plasmodium CSP polypeptide construct described herein optionally includes one or more of the following Plasmodium CSP polypeptide regions or portions (e.g., antigenic portions) thereof, and if present, are in the following N-terminus to C-terminus order: (i) one or more Plasmodium CSP N-terminal regions or portions (e.g., antigenic portions) thereof, (ii) one or more Plasmodium CSP N-terminal end regions or portions (e.g., antigenic portions) thereof, (iii) one or more Plasmodium CSP junction regions, portions (e.g., antigenic portions) thereof, or variants thereof, (iv) one or more repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (v) one or more Plasmodium CSP major repeat regions or portions (e.g., antigenic portions) thereof, and (vi) one or more Plasmodium CSP C-terminal regions
  • a Plasmodium CSP polypeptide construct described herein optionally includes one or more of the following Plasmodium CSP polypeptide regions or portions (e.g., antigenic portions) thereof, and if present, are in the following N-terminus to C-terminus order: (i) one Plasmodium CSP N-terminal region or portion (e.g., antigenic portion) thereof, (ii) one Plasmodium CSP N-terminal end region or portion (e.g., antigenic portion) thereof, (iii) one Plasmodium CSP junction region, portion (e.g., antigenic portion) thereof, or variant thereof, (iv) one or more Plasmodium CSP minor repeat sequences, (v) one Plasmodium CSP major repeat region or portion (e.g., antigenic portion) thereof, and (vi) one Plasmodium CSP C-terminal region or portion (e.g., antigenic portion) thereof.
  • a Plasmodium CSP polypeptide construct described herein includes one or more helper antigens.
  • helper antigens include those described in, e.g., WO2020128031 (which is incorporated herein by reference in its entirety) (e.g., P2 tetanus toxin, PADRE peptide, Hepatitis B surface antigen (HBsAg)).
  • a helper antigen is a malarial protein (e.g., a malarial protein described herein), provided that the antigen is not a CSP polypeptide or portion thereof.
  • a helper antigen is Plasmodium 2-phospho-D-glycerate hydro-lyase antigen, Plasmodium liver stage antigen 1(a), (LSA-1(a)), Plasmodium liver stage antigen 1(b) (LSA-1(b)), Plasmodium thrombospondin-related anonymous protein (TRAP), Plasmodium liver stage associated protein 1 (LSAP1), Plasmodium liver stage associated protein 2 (LSAP2), Plasmodium UIS3, Plasmodium UIS4, Plasmodium ETRAMP10.3, Plasmodium liver specific protein 1 (LISP-1), Plasmodium liver specific protein 2 (LISP-2), Plasmodium liver stage antigen 3 (LSA-3), Plasmodium EXP1, Plasmodium E140, Plasmodium reticulocyte-binding protein homolog 5 (Rh5), Plasmodium glutamic acid-rich protein (GARP), Plasmodium parasite-infected erythrocyte surface protein 2 (PIESP2)
  • a helper antigen comprises or consists of a P. falciparum 2-phospho-D-glycerate hydro-lyase antigen, e.g., comprising or consisting of amino acid sequence of ELDGSKNEWGWSKSKLGANA (SEQ ID NO: 388).
  • a helper antigen comprises or consists of a P.
  • falciparum liver-stage antigen 3 comprising or consisting of amino acid sequence of ENVQVSDELFNELLNSVDVNGEVKENILEESQVNDDIFNSLVKSVQQEQQHNVEEKVEESVEENDEESVEENVEENVEENDDESVAS SVEESIASSVDESIDSSIEENVAPTVEEIVAPTVEEIVAPSVVESVAPSVEESVEENVEESVAENVEESVAENVEESVAENV EESVAENVEESVA (SEQ ID NO: 391).
  • a helper antigen comprises or consists of an Anopheles antigen, e.g., an Anopheles gambiae TRIO, e.g., comprising or consisting of amino acid sequence of MCRGLSAVLILLVSLSAQLHVVVGEEAPKPEKEICGLKVGRLLDSVKGWLSVSQQEKCPLNKYCENKIQADQYNLVPLTCIRWRSLNP ASPTGSLGGKDVVSKIDAAMSNFKTLFEPMKADLAKLEEEVKRQVLDAWKALEPLQKEVYRSTLASGRIERAVFYSFMEMGDNVKLD NYFQPANVEELLKYAWALPMHKKQRSMYDLIGQLVQSSKSPMLQTLHAVELATVVNPELENRENLLNDQVVQLRDNLYKNSFATLV SIARHFPDHFDTLRQRLFKLPDGSKPGADTLPNIVNFIAQLPSDELRLSSVDLLLQSLTAENGTLVQDPEYVYRL
  • a Plasmodium CSP polypeptide construct described herein comprises a secretory signal (e.g., a secretory signal described herein) and a helper antigen immediately follows the secretory signal.
  • a Plasmodium CSP polypeptide construct described herein comprises a helper antigen located at the C-terminus.
  • a Plasmodium CSP polypeptide construct described herein comprises a linker between the CSP portion and the helper antigen. 3.
  • a Plasmodium CSP polypeptide construct described herein includes one or more multimerization regions (e.g., a heterologous multimerization region).
  • a heterologous multimerization region comprises a dimerization, trimerization or tetramerization region.
  • a multimerization region is one described in WO2017/081082, which is incorporated herein by reference in its entirety (e.g., SEQ ID NOs: 1116-1167, or fragments or variants thereof).
  • Exemplary trimerization and tetramerization regions include, but are not limited to, engineered leucine zippers, fibritin foldon domain from enterobacteria phage T4, GCN4pll, GCN4-pll, and p53.
  • a provided Plasmodium CSP polypeptide construct described herein is able to form a trimeric complex.
  • a provided Plasmodium CSP polypeptide construct may comprise a multimerization region allowing formation of a multimeric complex, such as for example a trimeric complex of a Plasmodium CSP polypeptide construct described herein.
  • a multimerization region allowing formation of a multimeric complex comprises a trimerization region, for example, a trimerization region described herein.
  • a Plasmodium CSP polypeptide construct includes a T4-fibritin-derived “foldon” trimerization region, for example, to increase its immunogenicity.
  • a Plasmodium CSP polypeptide construct includes a multimerization region comprising or consisting of the amino acid sequence GYIPEAPRDGQAYVRKDGEWVLLSTFLGRSLEVLFQGPG (SEQ ID NO: 399). 4.
  • a Plasmodium CSP polypeptide construct described herein includes one or more self-assembling regions (e.g., a self-assembling nanoparticle region, e.g., a heterologous self-assembling nanoparticle region).
  • a self-assembling nanoparticle region is a ferritin region.
  • a ferritin region is from H. pylori.
  • a ferritin region comprises or consists of a sequence according to the amino acid sequence of 402). 5.
  • a Plasmodium CSP polypeptide construct described herein includes one or more Plasmodium CSP polypeptide regions or portions thereof as described above. Exemplary combinations of regions are described below.
  • a Plasmodium CSP polypeptide construct described herein includes one or more regions or portions of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium CSP polypeptide construct described herein includes one or more regions or portions of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium CSP polypeptide construct described herein includes one or more of a N-terminal region, a N-terminal end region, a junction region, a minor repeat region, a major repeat region and a C-terminal region or corresponding portions thereof of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium CSP polypeptide construct described herein has the structure: N- terminal region – N-terminal end region – junction region – minor repeat region – major repeat region – C-terminal region, wherein the regions are from CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • such Plasmodium CSP polypeptide constructs have immediately following the C-terminal region a serine or a serine and a valine.
  • the N-terminal region or portion thereof comprises the amino acid sequence of positions 19 to 80 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 19 to 80 of SEQ ID NO: 1.
  • the N-terminal end region or portion thereof comprises the amino acid sequence of positions 81 to 92 of SEQ ID NO: 1, or the amino acid sequence of positions 81 to 92 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • the junction region or portion thereof comprises the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1, or the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • the minor repeat region or portion thereof comprises the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1.
  • the major repeat region or portion thereof comprises the amino acid sequence of positions 129 to 272 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 129 to 272 of SEQ ID NO: 1.
  • the C-terminal region or portion thereof comprises the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1.
  • a Plasmodium CSP polypeptide construct comprises the amino acid sequence of positions 19-375 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 19-375 of SEQ ID NO: 1.
  • a Plasmodium CSP polypeptide construct that includes all CSP regions as mentioned before and includes a serine or serine and valine immediately following the C-terminal region is referred to as a full-length CSP construct.
  • a Plasmodium CSP polypeptide construct can have the following structure: full-length CSP construct; Secretory signal (sec)-full-length CSP construct; full-length CSP construct-transmembrane region (TMD); sec-full-length CSP construct-TMD; Plasmodium falciparum (Pf) sec-full-length CSP construct; full-length CSP construct-PfTMD; or Pfsec-full-length CSP construct-PfTMD.
  • a Plasmodium CSP polypeptide construct described herein includes one or more of a N-terminal end region, a junction region, a minor repeat region, a major repeat region portion and a C-terminal region or corresponding portions thereof of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium CSP polypeptide construct described herein has the structure: N- terminal end region – junction region – [minor repeat region–major repeat region portion] x – minor repeat region– C- terminal region, wherein the [minor repeat region–major repeat region portion] repeats x times, and wherein the regions are from CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • x is 2 to 5 (i.e., the [minor repeat region–major repeat region portion] repeats 2 to 5 times).
  • such Plasmodium CSP polypeptide constructs have two repeats of a [minor repeat region– major repeat region portion], such that a Plasmodium CSP polypeptide construct described herein has the structure: N-terminal end region – junction region – minor repeat region – major repeat region portion – minor repeat region – major repeat region portion – minor repeat region – C-terminal region.
  • the N-terminal end region or portion thereof comprises the amino acid sequence of positions 81 to 92 of SEQ ID NO: 1, or the amino acid sequence of positions 81 to 92 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • the junction region includes an R1 region (amino acids 93-97) of SEQ ID NO:1.
  • the junction region or portion thereof comprises the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1, or the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • a minor repeat region or portion thereof comprises the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1.
  • a major repeat region portion comprises at least four repeats, at least five repeats, at least six repeats, at least seven repeats of the sequence NANP (SEQ ID NO: 230).
  • the major repeat region portion comprises a sequence of NANPNANPNANPNANPNANPNANPNANPNANPNANPNP (SEQ ID NO: 311).
  • the C-terminal region or portion thereof comprises the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1.
  • Plasmodium CSP polypeptide construct that includes all CSP regions or corresponding portions thereof as mentioned before and includes noncontiguous minor repeat regions (i.e., minor repeat region – major repeat region portion – minor repeat region – major repeat region portion – minor repeat region) is referred to as a 3xMR CSP construct.
  • a Plasmodium CSP polypeptide construct can have the following structure: 3xMR CSP construct; sec-3xMR CSP construct; 3xMR CSP construct-TMD; or sec-3xMR CSP construct-TMD.
  • a Plasmodium CSP polypeptide construct described herein includes one or more of a N-terminal end region, a junction region, a minor repeat region, a major repeat region and a C-terminal region or corresponding portions thereof of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium CSP polypeptide construct described herein has the structure: N- terminal end region – junction region – minor repeat region – major repeat region – C-terminal region, wherein the regions are from CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • such Plasmodium CSP polypeptide constructs have immediately following the C-terminal region a serine or a serine and a valine.
  • the N-terminal end region or portion thereof comprises the amino acid sequence of positions 81 to 92 of SEQ ID NO: 1, or the amino acid sequence of positions 81 to 92 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • the junction region or portion thereof comprises the amino acid sequence of positions 93 to 104 of SEQ ID NO:1, or the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • the minor repeat region or portion thereof comprises the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 105 to 128 of SEQ ID NO:1.
  • the major repeat region or portion thereof comprises the amino acid sequence of positions 129 to 272 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 129 to 272 of SEQ ID NO: 1.
  • the C-terminal region or portion thereof comprises the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1.
  • Plasmodium CSP polypeptide construct that includes all CSP regions or corresponding portions thereof as mentioned before except the N-terminal region or a portion thereof and includes a serine or serine and valine immediately following the C-terminal region is referred to as a dNT CSP construct.
  • a Plasmodium CSP polypeptide construct can have the following structure: dNT CSP construct; sec-dNT CSP construct; dNT CSP construct-TMD; or sec-dNT CSP construct-TMD.
  • a Plasmodium CSP polypeptide construct described herein includes one or more of a N-terminal end region, a junction region, one or more minor repeat region, and a C-terminal region or corresponding portions thereof of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium CSP polypeptide construct described herein has the structure: N-terminal end region – junction region – one or more minor repeat region – C-terminal region, wherein the regions are from CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • such Plasmodium CSP polypeptide constructs have immediately following the C-terminal region a serine or a serine and a valine.
  • the N-terminal end region or portion thereof comprises the amino acid sequence of positions 81 to 92 of SEQ ID NO: 1, or the amino acid sequence of positions 81 to 92 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • the junction region or portion thereof comprises the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1, or the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • the minor repeat region or portion thereof comprises the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1.
  • the C-terminal region or portion thereof comprises the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1.
  • such Plasmodium CSP polypeptide constructs have more than one minor repeat region, such as three minor repeat regions.
  • Plasmodium CSP polypeptide construct that includes all CSP regions or corresponding portions thereof as mentioned before except the N-terminal region and the major repeat region or corresponding portions thereof, has one or more minor repeat region and includes a serine or serine and valine immediately following the C-terminal region is referred to as a dNT-dmajor CSP construct.
  • a Plasmodium CSP polypeptide construct can have the following structure: dNT-dmajor CSP construct; sec-dNT-dmajor CSP construct; dNT-dmajor CSP construct-TMD; or sec-dNT-dmajor CSP construct-TMD.
  • a Plasmodium CSP polypeptide construct described herein includes one or more of a junction region, one or more minor repeat regions, a major repeat region and a C-terminal region or corresponding portions thereof of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium CSP polypeptide construct described herein has the structure: junction region – one or more minor repeat region – major repeat region - C-terminal region, wherein the regions are from CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • such Plasmodium CSP polypeptide constructs have immediately following the C-terminal region a serine or a serine and a valine.
  • the junction region or portion thereof comprises the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1, or the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • the minor repeat region or portion thereof comprises the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1.
  • the major repeat region or portion thereof comprises the amino acid sequence of positions 129 to 272 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 129 to 272 of SEQ ID NO: 1.
  • the C-terminal region or portion thereof comprises the amino acid sequence of positions 273 to 375 of SEQ ID NO:1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1.
  • such Plasmodium CSP polypeptide constructs have more than one minor repeat region, such as three minor repeat regions.
  • Such a Plasmodium CSP polypeptide construct that includes all CSP regions or corresponding portions thereof as mentioned before except the N-terminal domain (i.e., exclude the N-terminal region and the N-terminal end region) or a portion thereof, has one or more minor repeat regions and includes a serine or serine and valine immediately following the C-terminal region is referred to as a dND CSP construct.
  • a Plasmodium CSP polypeptide construct can have the following structure: dND CSP construct; sec-dND CSP construct; dND CSP construct-TMD; or sec-dND CSP construct-TMD.
  • a Plasmodium CSP polypeptide construct described herein includes a junction region, one or more minor repeat region, and a C-terminal region or corresponding portions thereof of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium CSP polypeptide construct described herein has the structure: junction region – one or more minor repeat region – C- terminal region, wherein the regions are from CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • such Plasmodium CSP polypeptide constructs have immediately following the C-terminal region a serine or a serine and a valine.
  • the junction region or portion thereof comprises the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1, or the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • the minor repeat region or portion thereof comprises the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1.
  • the C- terminal region or portion thereof comprises the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1.
  • such Plasmodium CSP polypeptide constructs have more than one minor repeat region, such as three minor repeat regions.
  • Such a Plasmodium CSP polypeptide construct that includes all CSP regions or corresponding portions thereof as mentioned before except the N-terminal domain (i.e. exclude the N-terminal region and the N-terminal end region) and the major repeat region or corresponding portions thereof, has one or more minor repeat region and includes a serine or serine and valine immediately following the C-terminal region is referred to as a dND-dmajor CSP construct.
  • a Plasmodium CSP polypeptide construct can have the following structure: dND-dmajor CSP construct; sec-dND-dmajor CSP construct; dND-dmajor CSP construct-TMD; or sec-dND-dmajor CSP construct-TMD. N-terminal Domain and Major Repeat Region Deleted CSP Constructs with Junction Region Variants or Portions [0509]
  • a Plasmodium CSP polypeptide construct described herein includes a junction region variant or junction region portion, one or more minor repeat regions, and a C-terminal region or corresponding portions thereof of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium CSP polypeptide construct described herein has the structure: junction region variant or junction region portion – one or more minor repeat region – C-terminal region, wherein the regions are from CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • such Plasmodium CSP polypeptide constructs have immediately following the C-terminal region a serine or a serine and a valine.
  • the junction region variant or portion thereof comprises the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • the junction region variant or portion thereof comprises the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1 having a K93A mutation, an L94A mutation, or both.
  • the junction region portion consists of a portion of the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1.
  • the junction region portion consists of the amino acid sequence of positions 97 to 104 of SEQ ID NO: 1.
  • the junction region portion comprises or consists of the amino acid sequence of positions 98 to 104 of SEQ ID NO: 1.
  • the minor repeat region or portion thereof comprises the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1.
  • the C-terminal region or portion thereof comprises the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1.
  • such Plasmodium CSP polypeptide constructs have more than one minor repeat region, such as three minor repeat regions.
  • Such a Plasmodium CSP polypeptide construct that includes all CSP regions or corresponding portions thereof as mentioned before except the N-terminal domain (i.e. exclude the N-terminal region and the N-terminal end region) and the major repeat region or corresponding portions thereof, has a junction region variant or junction region portion, has one or more minor repeat region and includes a serine or serine and valine immediately following the C-terminal region is referred to as a dND-dmajor-modJ CSP construct.
  • a Plasmodium CSP polypeptide construct can have the following structure: dND-dmajor-modJ CSP construct; sec-dND-dmajor-modJ CSP construct; dND-dmajor-modJ CSP construct-TMD; or sec-dND-dmajor-modJ CSP construct-TMD.
  • Major Repeat Region Portion and C-terminal Region Containing CSP Constructs [0510]
  • a Plasmodium CSP polypeptide construct described herein includes a major repeat region portion and a C-terminal region or corresponding portions thereof of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium CSP polypeptide construct described herein has the structure: major repeat region portion – C-terminal region, wherein the regions are from CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • such Plasmodium CSP polypeptide constructs have immediately following the C-terminal region a serine or a serine and a valine.
  • such Plasmodium CSP polypeptide constructs have between 2 and 35 repeats of the amino acid sequence NANP (SEQ ID NO: 230), preferably 18 repeats of the amino acid sequence NANP (SEQ ID NO: 230) as a major repeat portion.
  • the C-terminal region or portion thereof comprises the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 273 to 375 of SEQ ID NO: 1.
  • a Plasmodium CSP polypeptide construct that includes only a portion of the CSP major repeat region, a C-terminal region and includes a serine or serine and valine immediately following the C-terminal region, or that includes corresponding portions thereof as mentioned before, is referred to as a pmajor-CT CSP construct.
  • a Plasmodium CSP polypeptide construct can have the following structure: pmajor-CT CSP construct; sec-pmajor-CT CSP construct; pmajor-CT CSP construct-TMD; or sec- pmajor-CT CSP construct-TMD. N-terminal and C-terminal deleted CSP constructs with noncontiguous minor repeat regions [0511]
  • a Plasmodium CSP polypeptide construct described herein includes one or more of an N-terminal end region, a junction region, a minor repeat region, a major repeat region portion and a C-terminal region or corresponding portions thereof of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium CSP polypeptide construct described herein has the structure: [junction region – minor repeat region – major repeat region portion] x , wherein the [junction region – minor repeat region – major repeat region portion] repeats x times, and wherein the regions are from CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • x is 2 to 5 (i.e., the [junction region – minor repeat region–major repeat region portion] repeats 2 to 5 times).
  • such Plasmodium CSP polypeptide constructs have three repeats of a [junction region – minor repeat region – major repeat region portion], such that the Plasmodium CSP polypeptide construct described herein has the structure: junction region – minor repeat region – major repeat region portion – junction region – minor repeat region – major repeat region portion – junction region – minor repeat region – C-terminal region.
  • the N- terminal end region or portion thereof comprises the amino acid sequence of positions 81 to 92 of SEQ ID NO: 1, or the amino acid sequence of positions 81 to 92 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • the junction region includes an R1 region (amino acids 93-97) of SEQ ID NO: 1.
  • the junction region repeats twice.
  • the junction region or portion thereof comprises a 2x repeat of the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1, or the amino acid sequence of positions 93 to 104 of SEQ ID NO: 1 having 1, 2, 3, 4, or 5 amino acid substitutions.
  • a minor repeat region or portion thereof comprises the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1, or an amino acid sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the amino acid sequence of positions 105 to 128 of SEQ ID NO: 1.
  • a major repeat region portion comprises at least four repeats, at least five repeats, at least six repeats, at least seven repeats of the sequence NANP (SEQ ID NO: 230).
  • the major repeat region portion comprises a sequence of NANPNANPNANPNANPNANPNANPNANPNANPNP (SEQ ID NO: 311).
  • the malaria construct further comprises one or more linkers (e.g., glycine-serine linkers).
  • the malaria construct further comprises a linker (e.g., a glycine-serine linker) after each major repeat region portion sequence.
  • the malaria construct comprises a linker (e.g., a glycine-serine linker) after the last partial major repeat sequence.
  • a linker has the amino acid sequence GGSGGGGSGG (SEQ ID NO: 404).
  • a Plasmodium CSP polypeptide construct that includes all CSP regions or corresponding portions thereof as mentioned before and includes three repeats of a [junction region – minor repeat region – major repeat region portion] is referred to as a 3xMR-dNC CSP construct.
  • a Plasmodium CSP polypeptide construct can have the following structure: 3xMR-dNC CSP construct; sec-3xMR-dNC CSP construct; 3xMR-dNC CSP construct-TMD; or sec-3xMR-dNC CSP construct-TMD. 6 NANP and 18 NANP CSP Constructs [0513]
  • a Plasmodium CSP polypeptide construct described herein includes one or more regions or portions of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a Plasmodium CSP polypeptide construct described herein includes at least two repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), two to eighteen repeats of the amino acid sequence of NANP (SEQ ID NO: 230), and a Plasmodium CSP C-terminal region or portion thereof.
  • an “18NANP CSP Construct” includes at least: three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), eighteen repeats of the amino acid sequence of NANP (SEQ ID NO: 230), and a Plasmodium CSP C-terminal region or antigenic portion thereof.
  • the three repeats of the amino acid sequence of NANPNVDP form a Plasmodium CSP minor repeat region.
  • a Plasmodium minor repeat region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 226.
  • the eighteen repeats of the amino acid sequence of NANP (SEQ ID NO: 230) form an antigenic portion of the Plasmodium CSP major repeat region.
  • an antigenic portion of a Plasmodium CSP major repeat region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 308.
  • an 18NANP CSP Construct comprises a Plasmodium CSP C-terminal region.
  • a Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 238.
  • an 18NANP CSP Construct comprises a Plasmodium CSP C-terminal region variant.
  • a Plasmodium CSP C-terminal region variant comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 994.
  • an 18NANP CSP Construct comprises an antigenic portion of a Plasmodium CSP C-terminal region.
  • an antigenic portion of a Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 261-267.
  • an 18NANP CSP Construct comprises an antigenic portion of a Plasmodium CSP C-terminal region variant.
  • an antigenic portion of a Plasmodium CSP C-terminal region variant comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 992-993.
  • an 18NANP CSP Construct comprises a serine or serine and valine immediately following the C-terminal region.
  • an 18NANP CSP Construct comprises a Plasmodium CSP junction region.
  • a Plasmodium CSP junction region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 272.
  • an 18NANP CSP Construct comprises a Plasmodium CSP N-terminal end region.
  • a Plasmodium CSP N-terminal end region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 285.
  • an 18NANP CSP Construct comprises a Plasmodium CSP N-terminal region or antigenic portion thereof.
  • a Plasmodium CSP N-terminal region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 288.
  • an antigenic portion of a Plasmodium CSP N-terminal region comprises or consists of a Plasmodium CSP N-terminal start region.
  • a Plasmodium CSP N-terminal start region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 1010.
  • an 18NANP CSP Construct comprises one or more linkers. In some embodiments, one or more linkers are located between regions. In some embodiments, a linker is a cleavage linker. In some embodiments, a cleavage linker is positioned within an 18NANP CSP Construct between an N-terminal region or portion thereof and a C-terminal region or portion thereof.
  • a Plasmodium CSP polypeptide construct can have the following structure: 18NANP CSP Construct; Secretory Signal (Sec) - 18NANP CSP Construct; 18NANP CSP Construct – Transmembrane Domain (TMD); Sec - 18NANP CSP Construct – TMD; Plasmodium falciparum (Pf) Sec - 18NANP CSP Construct; 18NANP CSP Construct – Pf TMD Pf Sec - 18NANP CSP Construct Pf Sec - 18NANP CSP Construct – Heterologous TMD; and Pf Sec - 18NANP CSP Construct – HSV TMD.
  • a “6NANP CSP Construct” includes at least: three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), six repeats of the amino acid sequence of NANP (SEQ ID NO: 230), an asparagine-alanine positioned immediately following the six repeats of the amino acid sequence of NANP (SEQ ID NO: 230), and a Plasmodium CSP C-terminal region or antigenic fragment thereof (referred to herein as a “6NANP CSP Construct”).
  • the three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223) form a Plasmodium CSP minor repeat region.
  • a Plasmodium CSP minor repeat region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 407.
  • a major repeat region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 303.
  • a 6NANP CSP Construct comprises a Plasmodium CSP C-terminal region.
  • a Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 238.
  • an 6NANP CSP Construct comprises a Plasmodium CSP C-terminal region variant.
  • a Plasmodium CSP C-terminal region variant comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 994.
  • a 6NANP CSP Construct comprises an antigenic portion of a Plasmodium CSP C- terminal region.
  • an antigenic portion of a Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 261-267.
  • an 6NANP CSP Construct comprises an antigenic portion of a Plasmodium CSP C-terminal region variant.
  • an antigenic portion of a Plasmodium CSP C-terminal region variant comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 992-993.
  • a 6NANP CSP Construct comprises a serine or serine and valine immediately following the C-terminal region.
  • a 6NANP CSP Construct comprises a Plasmodium CSP junction region.
  • a Plasmodium CSP junction region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 272.
  • a 6NANP CSP Construct comprises a Plasmodium CSP N-terminal end region.
  • a Plasmodium CSP N-terminal end region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 285.
  • a 6NANP CSP Construct comprises a Plasmodium CSP N-terminal region or antigenic portion thereof.
  • a Plasmodium CSP N-terminal region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 288.
  • a 6NANP CSP Construct comprises one or more linkers. In some embodiments, one or more linkers are located between regions.
  • a Plasmodium CSP polypeptide construct can have the following structure: 6NANP CSP Construct; Sec - 6NANP CSP Construct; 6NANP CSP Construct – TMD; Sec - 6NANP CSP Construct – TMD; Pf Sec - 6NANP CSP Construct; 6NANP CSP Construct – Pf TMD; Pf Sec - 6NANP CSP Construct – Pf TMD; Pf Sec - 6NANP CSP Construct – Heterologous TMD; Pf Sec - 6NANP CSP Construct – HSV TMD; Heterologous Sec - 6NANP CSP Construct – Heterologous TMD; HSV Sec - 6NANP CSP Construct – HSV TMD; Heterologous Sec - 6NANP CSP Construct – HSV TMD; Heterologous Sec - 6NANP CSP Construct – HSV TMD; Heterologous Sec - 6NANP CSP Construct
  • a Plasmodium CSP polypeptide construct described herein includes one or more regions or portions of CSP from Plasmodium falciparum, preferably from Plasmodium falciparum isolate 3D7.
  • a “T-cell C-term CSP Construct” includes at least: a portion of a Plasmodium CSP C- terminal region, wherein the portion of the Plasmodium CSP C-terminal region comprises an amino acid sequence according to SEQ ID NO: 261 (YLX 3 X 4 IQX 5 SLST), wherein X 3 is N or K, X 4 is K, I, or R, and X 5 is N or Y.
  • a portion of a Plasmodium CSP C-terminal region comprises an amino acid sequence according to SEQ ID NO: 261-267.
  • a portion of a Plasmodium CSP C-terminal region comprises or consists of an amino acid sequence according to SEQ ID NO: 238.
  • a T-cell C-term CSP Construct comprises a serine or serine and valine immediately following the C-terminal region.
  • a T-cell C-term CSP Construct further comprises a Plasmodium CSP major repeat region or an antigenic portion thereof.
  • a T-cell C-term CSP Construct further comprises a Plasmodium CSP major repeat region.
  • a Plasmodium CSP major repeat region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 156.
  • a T-cell C-term CSP Construct comprises a portion of a Plasmodium CSP major repeat region.
  • a portion of the Plasmodium CSP major repeat region comprises or consists of six repeats of the amino acid sequence of NANP (SEQ ID NO: 230).
  • a portion of the Plasmodium CSP major repeat region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 303.
  • an asparagine-alanine is positioned immediately following the six repeats of the amino acid sequence of NANP.
  • a portion of the Plasmodium CSP major repeat region comprises or consists of 18 repeats of the amino acid sequence of NANP (SEQ ID NO: 230).
  • a portion of the Plasmodium CSP major repeat region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 308.
  • a T-cell C-term CSP Construct comprises a Plasmodium CSP minor repeat region.
  • a Plasmodium CSP minor repeat region comprises or consists of the three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223).
  • a Plasmodium CSP minor repeat region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 407.
  • a T-cell C-term CSP Construct comprises a Plasmodium CSP junction region.
  • a Plasmodium CSP junction region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 272.
  • a T-cell C-term CSP Construct comprises a Plasmodium CSP N-terminal end region.
  • a Plasmodium CSP N-terminal end region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 285.
  • a T-cell C-term CSP Construct comprises a Plasmodium CSP N-terminal region or antigenic portion thereof.
  • a Plasmodium CSP N-terminal region comprises or consists of an amino acid sequence having 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 100% identity to SEQ ID NO: 288.
  • a T-cell C-term CSP Construct comprises one or more linkers. In some embodiments, one or more linkers are located between regions.
  • a Plasmodium CSP polypeptide construct can have the following structure: T-cell C-term CSP Construct; Sec – T-cell C-term CSP Construct; T-cell C-term CSP Construct – TMD; Sec – T-cell C-term CSP Construct – TMD; Heterologous Sec - T-cell C-term CSP Construct – Multimerization Domain; or Heterologous Sec - T-cell C-term CSP Construct – Self-Assembly Domain.
  • a Plasmodium CSP polypeptide construct can have the following structure: Pf Sec – N-terminal Region – N-terminal End Region – Junction Region – Minor Repeat Region – Antigenic Portion of a Major Repeat Region – C-terminal Region; Pf Sec – N-terminal Region – N-terminal End Region – Junction Region – Minor Repeat Region – Antigenic Portion of a Major Repeat Region – C-terminal Region – Pf TMD; N-terminal Region – N-terminal End Region – Junction Region – Minor Repeat Region –Major Repeat Region – C-terminal Region Variant; Sec – N-terminal Region – N-terminal End Region – Junction Region – Minor Repeat Region – Major Repeat Region – C-terminal Region Variant; N-terminal Region – N-terminal End Region – Junction Region – Minor Repeat Region – Major Repeat Region – C-terminal Region Variant; N-terminal Region – N-terminal End Region – Junction Region – Minor
  • Plasmodium CSP polypeptide constructs described herein has an amino acid sequence provided in Table 4, and/or is encoded by a nucleotide sequence provided in Table 11.
  • Table 4 Amino Acid Sequences Encoding Exemplary Plasmodium CSP Polypeptide Constructs C.
  • Plasmodium polypeptide constructs as described herein e.g., Plasmodium T-cell string polypeptide constructs as described herein, Plasmodium CSP polypeptide constructs as described, or both
  • a Plasmodium polypeptide construct described herein includes a secretory signal, e.g., that is functional in mammalian cells.
  • a secretory signal comprises or consists of a Plasmodium secretory signal.
  • a Plasmodium secretory signal comprises or consists of a Plasmodium CSP secretory signal.
  • a Plasmodium CSP secretory signal is from Plasmodium falciparum.
  • a Plasmodium CSP secretory signal is from Plasmodium falciparum isolate 3D7 (SEQ ID NO.332).
  • a utilized secretory signal is a heterologous secretory signal.
  • a heterologous secretory signal comprises or consists of a non-human secretory signal.
  • a heterologous secretory signal comprises or consists of a viral secretory signal.
  • a viral secretory signal comprises or consists of an HSV secretory signal (e.g., an HSV-1 or HSV-2 secretory signal).
  • an HSV secretory signal comprises or consists of an HSV glycoprotein D (gD) secretory signal.
  • an HSV secretory signal comprises or consists of an HSV glycoprotein D (gD) secretory signal according to SEQ ID NO: 323 (MGGAAARLGAVILFVVIVGLHGVRG).
  • a secretory signal comprises or consists of an Ebola virus secretory signal.
  • an Ebola virus secretory signal comprises or consists of an Ebola virus spike glycoprotein (SGP) secretory signal.
  • SGP Ebola virus spike glycoprotein
  • a secretory signal is characterized by a length of about 15 to 30 amino acids.
  • a secretory signal is positioned at the N-terminus of a Plasmodium polypeptide construct described herein.
  • a secretory signal preferably allows transport of a Plasmodium polypeptide construct with which it is associated into a defined cellular compartment, preferably a cell surface, endoplasmic reticulum (ER) or endosomal-lysosomal compartment.
  • a secretory signal is selected from an S1S2 secretory signal (aa 1-19), an immunoglobulin secretory signal (aa 1-22), a human SPARC secretory signal, a human insulin isoform 1 secretory signal, a human albumin secretory signal, etc.
  • a Plasmodium polypeptide construct described herein does not comprise a secretory signal.
  • a secretory signal is one listed in Table 5, or a secretory signal having 1, 2, 3, 4, or 5 amino acid differences relative thereto.
  • a signal sequence is selected from those included in the Table 5 below and/or those encoded by the sequences in Table 6 below. Table 5: Exemplary secretory signals
  • Table 6 Exemplary polynucleotide sequences encoding secretory signals
  • a Plasmodium polypeptide construct described herein includes a transmembrane region (also referred to herein as a “transmembrane domain”).
  • a transmembrane region comprises or consists of a Plasmodium transmembrane region.
  • a utilized transmembrane region is one that is normally associated with CSP in nature.
  • a Plasmodium transmembrane region comprises or consists of a Plasmodium CSP glycosylphosphatidylinositol (GPI) anchor region, e.g., SEQ ID NO: 385.
  • GPI Plasmodium CSP glycosylphosphatidylinositol
  • a utilized transmembrane region is a heterologous transmembrane region.
  • Transmembrane regions are known in the art, any of which can be utilized in a Plasmodium polypeptide construct described herein.
  • a transmembrane region comprises or is a transmembrane domain of Hemagglutinin (HA) of Influenza virus, Env of HIV-1, equine infectious anaemia virus (EIAV), murine leukaemia virus (MLV), mouse mammary tumor virus, G protein of vesicular stomatitis virus (VSV), Rabies virus, or a seven transmembrane domain receptor.
  • HA Hemagglutinin
  • EIAV equine infectious anaemia virus
  • MMV murine leukaemia virus
  • VSV vesicular stomatitis virus
  • Rabies virus or a seven transmembrane domain receptor.
  • a heterologous transmembrane region does not comprise a hemagglutinin transmembrane region.
  • a heterologous transmembrane region comprises or consists of a non-human transmembrane region.
  • a heterologous transmembrane region comprises or consists of a viral transmembrane region.
  • a heterologous transmembrane region comprises or consists of an HSV transmembrane region, e.g., an HSV-1 or HSV-2 transmembrane region.
  • an HSV transmembrane region comprises or consists of an HSV gD transmembrane region, e.g., comprising or consisting of an amino acid sequence of GLIAGAVGGSLLAALVICGIVYWMRRHTQKAPKRIRLPHIR (SEQ ID NO: 379).
  • a heterologous transmembrane region comprises or consists of a human transmembrane region.
  • a human transmembrane region comprises or consists of a human decay accelerating factor glycosylphosphatidylinositol (hDAF-GPI) anchor region.
  • an hDAF- GPI anchor region comprises or consists of an amino acid sequence of PNKGSGTTSGTTRLLSGHTCFTLTGLLGTLVTMGLLT (SEQ ID NO: 382).
  • a transmembrane region is located at the N-terminus of a Plasmodium polypeptide construct. In some embodiments, a transmembrane region is located at the C-terminus of a Plasmodium polypeptide construct. In some embodiments, a transmembrane region is not located at the N-terminus or C- terminus of a Plasmodium polypeptide construct.
  • a Plasmodium polypeptide construct described herein does not comprise a transmembrane region. 3.
  • Linkers [0568] In some embodiments, a Plasmodium polypeptide construct described herein includes one or more linkers. In some embodiments, a linker is or comprises 2, 3, 4, 5, 6, 7, 8, 9, 10 or more amino acids. In some embodiments, a linker is or comprises no more than about 30, 25, 20, 15, 10 or fewer amino acids. A linker can include any amino acid sequence and is not limited to any particular amino acids. In some embodiments, a linker comprises one or more glycine (G) amino acids. In some embodiments, a linker comprises one or more serine (S) amino acids.
  • G glycine
  • S serine
  • a linker comprises a glycine-serine linker.
  • a “glycine-serine linker” as used herein refers to a linker that comprises predominantly (e.g., 80% or more) glycine and serine amino acids.
  • a linker includes amino acids selected based on a cleavage predictor to generate highly-cleavable linkers.
  • a linker is or comprises S-G4-S-G4-S (SEQ ID NO: 405).
  • a linker is or comprises GSPGSGSGS (SEQ ID NO: 455).
  • a linker is or comprises GGSGGGGSGG (SEQ ID NO: 404). In some embodiments, a linker is or comprises AGNRVRRSVG (SEQ ID NO: 412). In some embodiments, a linker is one presented in Table 7. In some embodiments, a linker is or comprises a sequence as set forth in WO2017/081082, which is incorporated herein by reference in its entirety (see SEQ ID NOs: 1509-1565, or a fragment or variant thereof). [0570] In some embodiments, a Plasmodium T-cell string polypeptide construct described herein comprises a linker between two Plasmodium T-cell antigens or between two or more antigenic polypeptide fragments from the same T-cell antigen.
  • a Plasmodium CSP polypeptide construct described herein comprises a linker between a C-terminal region or portion thereof and a transmembrane region. In some embodiments, a Plasmodium CSP polypeptide construct described herein comprises a linker after a minor repeat sequence. In some embodiments, a Plasmodium CSP polypeptide construct described herein comprises a linker after a major repeat sequence or portion thereof. [0571] Exemplary linkers are provided in the following Table 7: Table 7: Exemplary linkers D.
  • Polyribonucleotide Constructs [0572] Polyribonucleotides described herein encode one or more constructs (e.g., one or more Plasmodium T- cell string polypeptide constructs, one or more Plasmodium CSP polypeptide constructs, or both) as described herein. In some embodiments, polyribonucleotides described herein can be included in an RNA construct. In some embodiments, an RNA construct provided herein comprises a nucleotide sequence that encodes a 5’UTR and/or a 3’ UTR. In some embodiments, polyribonucleotides described herein can comprise a ribonucleotide sequence that encodes a polyA tail.
  • polyribonucleotides described herein may comprise a 5’ cap, which may be incorporated during transcription, or joined to a polyribonucleotide post-transcription.
  • 5 A structural feature of mRNAs is cap structure at five-prime end (5’).
  • Natural eukaryotic mRNA comprises a 7-methylguanosine cap linked to the mRNA via a 5 ⁇ to 5 ⁇ -triphosphate bridge resulting in cap0 structure (m 7 GpppN).
  • RNA capping is well researched and is described, e.g., in Decroly E et al. (2012) Nature Reviews 10: 51- 65; and in Ramanathan A. et al., (2016) Nucleic Acids Res; 44(16): 7511–7526, the entire contents of each of which is hereby incorporated by reference.
  • a 5’-cap structure which may be suitable in the context of the present invention is a cap0 (methylation of the first nucleobase, e.g., m 7 GpppN), cap1 (additional methylation of the ribose of the adjacent nucleotide of m 7 GpppN), cap2 (additional methylation of the ribose of the 2nd nucleotide downstream of the m 7 GpppN), cap3 (additional methylation of the ribose of the 3rd nucleotide downstream of the m 7 GpppN), cap4 (additional methylation of the ribose of the 4th nucleotide downstream of the m 7 GpppN), ARCA (“anti-reverse cap analogue”), modified ARCA (e.g.
  • RNA e.g., mRNA
  • 5'-cap refers to a structure found on the 5'-end of an RNA, e.g., mRNA, and generally includes a guanosine nucleotide connected to an RNA, e.g., mRNA, via a 5'- to 5'-triphosphate linkage (also referred to as Gppp or G(5')ppp(5')).
  • a guanosine nucleoside included in a 5’ cap may be modified, for example, by methylation at one or more positions (e.g., at the 7-position) on a base (guanine), and/or by methylation at one or more positions of a ribose.
  • a guanosine nucleoside included in a 5’ cap comprises a 3’O methylation at a ribose (3’OMeG).
  • a guanosine nucleoside included in a 5’ cap comprises methylation at the 7-position of guanine (m 7 G).
  • a guanosine nucleoside included in a 5’ cap comprises methylation at the 7-position of guanine and a 3’ O methylation at a ribose (m 7 (3’OMeG)).
  • m 7 (3’OMeG) a ribose that is used in the above paragraph, e.g., “(m 2 7,3’-O )G” or “m 7 (3’OMeG)”, applies to other structures described herein.
  • providing an RNA with a 5'-cap disclosed herein may be achieved by in vitro transcription, in which a 5'-cap is co-transcriptionally expressed into an RNA strand, or may be attached to an RNA post-transcriptionally using capping enzymes.
  • co-transcriptional capping with a cap disclosed improves the capping efficiency of an RNA compared to co-transcriptional capping with an appropriate reference comparator.
  • improving capping efficiency can increase a translation efficiency and/or translation rate of an RNA, and/or increase expression of an encoded polypeptide.
  • alterations to polynucleotides generates a non-hydrolyzable cap structure which can, for example, prevent decapping and increase RNA half-life.
  • a utilized 5’ caps is a cap0, a cap1, or cap2 structure. See, e.g., Fig. 1 of Ramanathan A et al., and Fig.
  • an RNA described herein comprises a cap1 structure. In some embodiments, an RNA described herein comprises a cap2. [0578] In some embodiments, an RNA described herein comprises a cap0 structure. In some embodiments, a cap0 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m 7 )G).
  • such a cap0 structure is connected to an RNA via a 5'- to 5'-triphosphate linkage and is also referred to herein as (m 7 )Gppp.
  • a cap0 structure comprises a guanosine nucleoside methylated at the 2’-position of the ribose of guanosine.
  • a cap0 structure comprises a guanosine nucleoside methylated at the 3’-position of the ribose of guanosine.
  • a guanosine nucleoside included in a 5’ cap comprises methylation at the 7-position of guanine and at the 2’-position of the ribose ((m 2 7,2’-O )G). In some embodiments, a guanosine nucleoside included in a 5’ cap comprises methylation at the 7-position of guanine and at the 2’-position of the ribose ((m 2 7,3’-O )G).
  • a cap1 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m 7 )G) and optionally methylated at the 2’ or 3’ position of the ribose, and a 2’O methylated first nucleotide in an RNA ((m 2’-O )N 1 ).
  • a cap1 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m 7 )G) and the 3’ position of the ribose, and a 2’O methylated first nucleotide in an RNA ((m 2’-O )N 1 ).
  • a cap1 structure is connected to an RNA via a 5'- to 5'- triphosphate linkage and is also referred to herein as, e.g., ((m 7 )Gppp( 2'-O )N 1 ) or (m 2 7,3’-O )Gppp( 2'-O )N 1 ), wherein N 1 is as defined and described herein.
  • a cap1 structure comprises a second nucleotide, N 2 , which is at position 2 and is chosen from A, G, C, or U, e.g., (m 7 )Gppp( 2'-O )N 1 pN 2 or (m 2 7,3’-O )Gppp( 2'-O )N 1 pN 2 , wherein each of N 1 and N 2 is as defined and described herein.
  • a cap2 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m 7 )G) and optionally methylated at the 2’ or 3’ position of the ribose, and a 2’O methylated first and second nucleotides in an RNA ((m 2’-O )N 1 p(m 2’-O )N 2 ).
  • a cap2 structure comprises a guanosine nucleoside methylated at the 7-position of guanine ((m 7 )G) and the 3’ position of the ribose, and a 2’O methylated first and second nucleotide in an RNA.
  • a cap2 structure is connected to an RNA via a 5'- to 5'- triphosphate linkage and is also referred to herein as, e.g., ((m 7 )Gppp( 2'-O )N 1 p( 2'-O )N 2 ) or (m 2 7,3’-O )Gppp( 2'-O )N 1 p( 2'- O )N 2 ), wherein each of N 1 and N 2 is as defined and described herein.
  • the 5’ cap is a dinucleotide cap structure.
  • the 5’ cap is a dinucleotide cap structure comprising N 1 , wherein N 1 is as defined and described herein.
  • the 5’ cap is a dinucleotide cap G*N 1 , wherein N 1 is as defined above and herein, and G* comprises a structure of formula (I): or a salt thereof, wherein each R 2 3 d X is O o [0582]
  • R 2 is -OH.
  • R 2 is -OCH 3 .
  • R 3 is - OH.
  • R 3 is -OCH 3 .
  • R 2 is -OH and R 3 is -OH.
  • R 2 is -OH and R 3 is -CH 3 . In some embodiments, R 2 is -CH 3 and R 3 is -OH. In some embodiments, R 2 is -CH 3 and R 3 is - CH 3 . [0583] In some embodiments, X is O. In some embodiments, X is S.
  • the 5’ cap is a dinucleotide cap0 structure (e.g., (m 7 )GpppN 1 , (m 2 7,2’-O )GpppN 1 , (m 2 7,3’-O )GpppN 1 , (m 7 )GppSpN 1 , (m 2 7,2’-O )GppSpN 1 , or (m 2 7,3’-O )GppSpN 1 ), wherein N 1 is as defined and described herein.
  • N 1 is as defined and described herein.
  • the 5’ cap is a dinucleotide cap0 structure (e.g., (m 7 )GpppN 1 , (m 2 7,2’-O )GpppN 1 , (m 2 7,3’- O )GpppN 1 , (m 7 )GppSpN 1 , (m 2 7,2’-O )GppSpN 1 , or (m 2 7,3’-O )GppSpN 1 ), wherein N 1 is G.
  • a dinucleotide cap0 structure e.g., (m 7 )GpppN 1 , (m 2 7,2’-O )GpppN 1 , (m 2 7,3’- O )GpppN 1 , wherein N 1 is G.
  • the 5’ cap is a dinucleotide cap0 structure (e.g., (m 7 )GpppN 1 , (m 2 7,2’-O )GpppN 1 , (m 2 7,3’-O )GpppN 1 , (m 7 )GppSpN 1 , (m 2 7,2’- O )GppSpN 1 , or (m 2 7,3’-O )GppSpN 1 ), wherein N 1 is A, U, or C.
  • a dinucleotide cap0 structure e.g., (m 7 )GpppN 1 , (m 2 7,2’-O )GpppN 1 , (m 2 7,3’-O )GpppN 1 , wherein N 1 is A, U, or C.
  • the 5’ cap is a dinucleotide cap1 structure (e.g., (m 7 )Gppp(m 2’-O )N 1 , (m 2 7,2’-O )Gppp(m 2’-O )N 1 , (m 2 7,3’-O )Gppp(m 2’-O )N 1 , (m 7 )GppSp(m 2’-O )N 1 , (m 2 7,2’- O )GppSp(m 2’-O )N 1 , or (m 2 7,3’-O )GppSp(m 2’-O )N 1 ), wherein N 1 is as defined and described herein.
  • N 1 is as defined and described herein.
  • the 5’ cap is selected from the group consisting of (m 7 )GpppG (“Ecap0”), (m 7 )Gppp(m 2’-O )G (“Ecap1”), (m 2 7,3’- O )GpppG (“ARCA” or “D1”), and (m 2 7,2’-O )GppSpG (“beta-S-ARCA”).
  • the 5’ cap is (m 7 )GpppG (“Ecap0”), having a structure: or a salt thereof.
  • the 5’ cap is (m 7 )Gppp(m 2’-O )G (“Ecap1”), having a structure: or a salt thereof.
  • the 5’ cap is (m 2 7,3’-O )GpppG (“ARCA” or “D1”), having a structure: or a salt thereof.
  • the 5’ cap is (m 2 7,3’-O )GppSpG (“beta-S-ARCA”), having a structure:
  • the 5’ cap is a trinucleotide cap structure. In some embodiments, the 5’ cap is a trinucleotide cap structure comprising N 1 pN 2 , wherein N 1 and N 2 are as defined and described herein. In some embodiments, the 5’ cap is a dinucleotide cap G*N 1 pN 2 , wherein N 1 and N 2 are as defined above and herein, and G* comprises a structure of formula (I): or a salt thereof, wherein R 2 , R 3 , and X are as defined and described herein.
  • the 5’ cap is a trinucleotide cap0 structure (e.g. (m 7 )GpppN 1 pN 2 , (m 2 7,2’- O )GpppN 1 pN 2 , or (m 2 7,3’-O )GpppN 1 pN 2 ), wherein N 1 and N 2 are as defined and described herein).
  • the 5’ cap is a trinucleotide cap1 structure (e.g., (m 7 )Gppp(m 2’-O )N 1 pN 2 , (m 2 7,2’-O )Gppp(m 2’-O )N 1 pN 2 , (m 2 7,3’-O )Gppp(m 2’-O )N 1 pN 2 ), wherein N 1 and N 2 are as defined and described herein.
  • the 5’ cap is a trinucleotide cap2 structure (e.g., (m 7 )Gppp(m 2’-O )N 1 p(m 2’-O )N 2 , (m 2 7,2’-O )Gppp(m 2’-O )N 1 p(m 2’-O )N 2 , (m 2 7,3’- O )Gppp(m 2’-O )N 1 p(m 2’-O )N 2 ), wherein N 1 and N 2 are as defined and described herein.
  • the 5’ cap is selected from the group consisting of (m2 7,3’-O )Gppp(m 2’-O )ApG (“CleanCap AG”, “CC413”), (m2 7,3’-O )Gppp(m 2’- O )GpG (“CleanCap GG”), (m 7 )Gppp(m 2’-O )ApG, (m 7 )Gppp(m 2’-O )G, (m 2 7,3’-O )Gppp(m 2 6,2’-O )ApG, and (m 7 )Gppp(m 2’- O )ApU.
  • the 5’ cap is (m 2 7,3’-O )Gppp(m 2’-O )ApG (“CleanCap AG”, “CC413”), having a structure:
  • the 5’ cap is (m 2 7,3’-O )Gppp(m 2’-O )GpG (“CleanCap GG”), having a structure: or a salt thereof.
  • the 5’ cap is (m 7 )Gppp(m 2’-O )ApG, having a structure:
  • the 5’ cap is (m 7 )Gppp(m 2’-O )GpG, having a structure: or a salt thereof.
  • the 5’ cap is (m 2 7,3’-O )Gppp(m 2 6,2’-O )ApG, having a structure:
  • the 5’ cap is (m 7 )Gppp(m 2’-O )ApU, having a structure: or a salt thereof.
  • the 5’ cap is a tetranucleotide cap structure.
  • the 5’ cap is a tetranucleotide cap structure comprising N 1 pN 2 pN 3 , wherein N 1 , N 2 , and N 3 are as defined and described herein.
  • the 5’ cap is a tetranucleotide cap G*N 1 pN 2 pN 3 , wherein N 1 , N 2 , and N 3 are as defined above and herein, and G* comprises a structure of formula (I): or a salt thereof, wherein R 2 , R 3 , and X are as defined and described herein.
  • the 5’ cap is a tetranucleotide cap0 structure (e.g.
  • the 5’ cap is a tetranucleotide Cap1 structure (e.g., (m 7 )Gppp(m 2’-O )N 1 pN 2 pN 3 , (m 2 7,2’-O )Gppp(m 2’- O )N 1 pN 2 pN 3 , (m 2 7,3’-O )Gppp(m 2’-O )N 1 pN 2 N 3 ), wherein N 1 , N 2 , and N 3 are as defined and described herein.
  • tetranucleotide Cap1 structure e.g., (m 7 )Gppp(m 2’-O )N 1 pN 2 pN 3 , (m 2 7,2’-O )Gppp(m 2’- O )N 1 pN 2 pN 3 , (m 2 7,3’-O )Gppp(m 2’-O )N 1 pN 2 N 3 ), wherein N 1
  • the 5’ cap is a tetranucleotide Cap2 structure (e.g., (m 7 )Gppp(m 2’-O )N 1 p(m 2’-O )N 2 pN 3 , (m 2 7,2’- O )Gppp(m 2’-O )N 1 p(m 2’-O )N 2 pN 3 , (m 2 7,3’-O )Gppp(m 2’-O )N 1 p(m 2’-O )N 2 pN 3 ), wherein N 1 , N 2 , and N 3 are as defined and described herein.
  • N 1 , N 2 , and N 3 are as defined and described herein.
  • the 5’ cap is selected from the group consisting of (m 2 7,3’-O )Gppp(m 2’- O )Ap(m 2’-O )GpG, (m 2 7,3’-O )Gppp(m 2’-O )Gp(m 2’-O )GpC, (m 7 )Gppp(m 2’-O )Ap(m 2’-O )UpA, and (m 7 )Gppp(m 2’-O )Ap(m 2’-O )GpG.
  • the 5’ cap is (m 2 7,3’-O )Gppp(m 2’-O )Ap(m 2’-O )GpG, having a structure: or a salt thereof.
  • the 5’ cap is (m 2 7,3’-O )Gppp(m 2’-O )Gp(m 2’-O )GpC, having a structure:
  • the 5’ cap is (m 7 )Gppp(m 2’-O )Ap(m 2’-O )UpA, having a structure: or a salt thereof.
  • the 5’ cap is (m 7 )Gppp(m 2’-O )Ap(m 2’-O )GpG, having a structure:
  • a 5 UTR utilized in accordance with the present disclosure comprises a cap proximal sequence, e.g., as disclosed herein.
  • a cap proximal sequence comprises a sequence adjacent to a 5’ cap.
  • a cap proximal sequence comprises nucleotides in positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide.
  • a cap structure comprises one or more polynucleotides of a cap proximal sequence.
  • a cap structure comprises an m 7 Guanosine cap and nucleotide +1 (N 1 ) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m 7 Guanosine cap and nucleotide +2 (N 2 ) of an RNA polynucleotide. In some embodiments, a cap structure comprises an m 7 Guanosine cap and nucleotides +1 and +2 (N 1 and N 2 ) of an RNA polynucleotide.
  • a cap structure comprises an m 7 Guanosine cap and nucleotides +1, +2, and +3 (N 1 , N 2 , and N 3 ) of an RNA polynucleotide.
  • one or more residues of a cap proximal sequence may be included in an RNA by virtue of having been included in a cap entity (e.g., a cap1 or cap2 structure, etc.); alternatively, in some embodiments, at least some of the residues in a cap proximal sequence may be enzymatically added (e.g., by a polymerase such as a T7 polymerase).
  • the 5’ cap is a dinucleotide cap structure, wherein the cap proximal sequence comprises N 1 of the 5’ cap, where N 1 is any nucleotide, e.g., A, C, G or U.
  • the 5’ cap is a trinucleotide cap structure (e.g., the trinucleotide cap structures described above and herein), wherein the cap proximal sequence comprises N 1 and N 2 of the 5’ cap, wherein N 1 and N 2 are independently any nucleotide, e.g., A, C, G or U.
  • the 5’ cap is a tetranucleotide cap structure (e.g., the trinucleotide cap structures described above and herein), wherein the cap proximal sequence comprises N 1 , N 2 , and N 3 of the 5’ cap, wherein N 1 , N 2 , and N 3 are any nucleotide, e.g., A, C, G or U.
  • a cap proximal sequence comprises N 1 of a the 5’ cap, and N 2 , N 3 , N and N 5 , wherein N 1 to N 5 correspond to positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide.
  • a cap proximal sequence comprises N 1 and N 2 of a the 5’ cap, and N 3 , N 4 and N 5 , wherein N 1 to N 5 correspond to positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide.
  • a cap proximal sequence comprises N 1 , N 2 , and N 3 of a the 5’ cap, and N 4 and N 5 , wherein N 1 to N 5 correspond to positions +1, +2, +3, +4, and/or +5 of an RNA polynucleotide.
  • N 1 is A.
  • N 1 is C.
  • N 1 is G.
  • N 1 is U.
  • N 2 is A.
  • N 2 is C.
  • N 2 is G.
  • N 2 is U.
  • N 3 is A. In some embodiments, N 3 is C. In some embodiments, N 3 is G. In some embodiments, N 3 is U. In some embodiments, N 4 is A. In some embodiments, N 4 is C. In some embodiments, N 4 is G. In some embodiments, N 4 is U. In some embodiments, N 5 is A. In some embodiments, N 5 is C. In some embodiments, N 5 is G. In some embodiments, N 5 is U. It will be understood that, each of the embodiments described above and herein (e.g., for N 1 through N 5 ) may be taken singly or in combination and/or may be combined with other embodiments of variables described above and herein (e.g., 5’ caps).
  • a nucleic acid utilized in accordance with the present disclosure comprises a 5'-UTR.
  • 5’-UTR may comprise a plurality of distinct sequence elements; in some embodiments, such plurality may be or comprise multiple copies of one or more particular sequence elements (e.g., as may be from a particular source or otherwise known as a functional or characteristic sequence element).
  • a 5’ UTR comprises multiple different sequence elements.
  • untranslated region or “UTR” is commonly used in the art to a region in a DNA molecule which is transcribed but is not translated into an amino acid sequence, or to the corresponding region in an RNA polynucleotide, such as an mRNA molecule.
  • An untranslated region (UTR) can be present 5' (upstream) of an open reading frame (5'-UTR) and/or 3' (downstream) of an open reading frame (3'-UTR).
  • the terms “five prime untranslated region” or “5' UTR” refer to a sequence of a polyribonucleotide between the 5' end of the polyribonucleotide (e.g., a transcription start site) and a start codon of a coding region of the polyribonucleotide.
  • “5' UTR” refers to a sequence of a polyribonucleotide that begins at the 5' end of the polyribonucleotide (e.g., a transcription start site) and ends one nucleotide (nt) before a start codon (usually AUG) of a coding region of the polyribonucleotide, e.g., in its natural context.
  • a 5' UTR comprises a Kozak sequence.
  • a 5'-UTR is downstream of the 5'-cap (if present), e.g., directly adjacent to the 5'-cap.
  • a 5’ UTR disclosed herein comprises a cap proximal sequence, e.g., as defined and described herein.
  • a cap proximal sequence comprises a sequence adjacent to a 5’ cap.
  • Exemplary 5’ UTRs include a human alpha globin (hAg) 5’UTR or a fragment thereof, a TEV 5’ UTR or a fragment thereof, a HSP705’ UTR or a fragment thereof, or a c-Jun 5’ UTR or a fragment thereof.
  • an RNA disclosed herein comprises a hAg 5’ UTR or a fragment thereof.
  • an RNA disclosed herein comprises a 5’ UTR having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a 5’ UTR with the sequence AGAAUAAACUAGUAUUCUUCUGGUCCCCACAGACUCAGAGAACCCGCCACC (SEQ ID NO: 563).
  • an RNA disclosed herein comprises a 5’ UTR having the sequence AGAAUAAACUAGUAUUCUUCUGGUCCCCACAGACUCAGAGAGAACCCGCCACC (SEQ ID NO: 563).
  • a polynucleotide e.g., DNA, RNA
  • a polyA sequence is situated downstream of a 3'- UTR, e.g., adjacent to a 3'-UTR.
  • poly(A) sequence or “poly-A tail” refers to an uninterrupted or interrupted sequence of adenylate residues which is typically located at the 3'-end of an RNA polynucleotide.
  • Poly(A) sequences are known to those of skill in the art and may follow the 3’-UTR in the RNAs described herein.
  • An uninterrupted poly(A) sequence is characterized by consecutive adenylate residues. In nature, an uninterrupted poly(A) sequence is typical.
  • polynucleotides disclosed herein comprise an uninterrupted Poly(A) sequence.
  • polynucleotides disclosed herein comprise interrupted Poly(A) sequence.
  • RNAs disclosed herein can have a poly(A) sequence attached to the free 3'-end of the RNA by a template- independent RNA polymerase after transcription or a poly(A) sequence encoded by DNA and transcribed by a template-dependent RNA polymerase.
  • a poly(A) sequence of about 120 A nucleotides has a beneficial influence on the levels of RNA in transfected eukaryotic cells, as well as on the levels of protein that is translated from an open reading frame that is present upstream (5’) of the poly(A) sequence (Holtkamp et al., 2006, Blood, vol. 108, pp. 4009-4017, which is herein incorporated by reference).
  • a poly(A) sequence in accordance with the present disclosure is not limited to a particular length; in some embodiments, a poly(A) sequence is any length.
  • a poly(A) sequence comprises, essentially consists of, or consists of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 A nucleotides, and, in particular, about 120 A nucleotides.
  • nucleotides in the poly(A) sequence typically at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% by number of nucleotides in the poly(A) sequence are A nucleotides, but permits that remaining nucleotides are nucleotides other than A nucleotides, such as U nucleotides (uridylate), G nucleotides (guanylate), or C nucleotides (cytidylate).
  • consists of means that all nucleotides in the poly(A) sequence, i.e., 100% by number of nucleotides in the poly(A) sequence, are A nucleotides.
  • a nucleotide or “A” refers to adenylate.
  • a poly(A) sequence is attached during RNA transcription, e.g., during preparation of in vitro transcribed RNA, based on a DNA template comprising repeated dT nucleotides (deoxythymidylate) in the strand complementary to the coding strand.
  • the DNA sequence encoding a poly(A) sequence (coding strand) is referred to as poly(A) cassette.
  • the poly(A) cassette present in the coding strand of DNA essentially consists of dA nucleotides but is interrupted by a random sequence of the four nucleotides (dA, dC, dG, and dT). Such random sequence may be 5 to 50, 10 to 30, or 10 to 20 nucleotides in length.
  • a cassette is disclosed in WO 2016/005324 A1, hereby incorporated by reference. Any poly(A) cassette disclosed in WO 2016/005324 A1 may be used in accordance with the present disclosure.
  • a poly(A) cassette that essentially consists of dA nucleotides but is interrupted by a random sequence having an equal distribution of the four nucleotides (dA, dC, dG, dT) and having a length of e.g., 5 to 50 nucleotides shows, on DNA level, constant propagation of plasmid DNA in E. coli and is still associated, on RNA level, with the beneficial properties with respect to supporting RNA stability and translational efficiency is encompassed.
  • the poly(A) sequence contained in an RNA polynucleotide described herein essentially consists of A nucleotides but is interrupted by a random sequence of the four nucleotides (A, C, G, U).
  • Such random sequence may be 5 to 50, 10 to 30, or 10 to 20 nucleotides in length.
  • no nucleotides other than A nucleotides flank a poly(A) sequence at its 3'-end, i.e., the poly(A) sequence is not masked or followed at its 3'-end by a nucleotide other than A.
  • the poly(A) sequence may comprise at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 nucleotides.
  • the poly(A) sequence may essentially consist of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 nucleotides. In some embodiments, the poly(A) sequence may consist of at least 20, at least 30, at least 40, at least 80, or at least 100 and up to 500, up to 400, up to 300, up to 200, or up to 150 nucleotides. In some embodiments, the poly(A) sequence comprises at least 100 nucleotides. In some embodiments, the poly(A) sequence comprises about 150 nucleotides. In some embodiments, the poly(A) sequence comprises about 120 nucleotides.
  • a polyA tail comprises a specific number of Adenosines, such as about 50 or more, about 60 or more, about 70 or more, about 80 or more, about 90 or more, about 100 or more, about 120, or about 150 or about 200.
  • a polyA tail of a string construct may comprise 200 A residues or less.
  • a polyA tail of a string construct may comprise about 200 A residues.
  • a polyA tail of a string construct may comprise 180 A residues or less.
  • a polyA tail of a string construct may comprise about 180 A residues.
  • a polyA tail may comprise 150 residues or less.
  • RNA comprises a poly(A) sequence comprising the nucleotide sequence of AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 569), or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA (SEQ ID NO: 569).
  • a poly(A) tail comprises a plurality of A residues interrupted by a linker.
  • a linker comprises the nucleotide sequence GCAUAUGAC (SEQ ID NO: 414).
  • 3' UTR [0622]
  • an RNA utilized in accordance with the present disclosure comprises a 3'-UTR.
  • the terms “three prime untranslated region,” “3' untranslated region,” or “3' UTR” refer to a sequence of an mRNA molecule that begins following a stop codon of a coding region of an open reading frame sequence. In some embodiments, the 3' UTR begins immediately after a stop codon of a coding region of an open reading frame sequence, e.g., in its natural context.
  • the 3' UTR does not begin immediately after stop codon of the coding region of an open reading frame sequence, e.g., in its natural context.
  • an RNA disclosed herein comprises a 3’ UTR with the sequence of CUGGUACUGCAUGCACGCAAUGCUAGCUGCCCCUUUCCCGUCCUGGGUACCCCGAGUCUCCCCCGACCUCGGGUCCCAGGUA UGCUCCCACCUCCACCUGCCCCACUCACCACCUCUGCUAGUUCCAGACACCUCCCAAGCACGCAGCAAUGCAGCUCAAAACGCU UAGCCUAGCCACACCCCCACGGGAAACAGCAGUGAUUAACCUUUAGCAAUAAACGAAAGUUUAACUAAGCUAUACUAACCCCA GGGUUGGUCAAUUUCGUGCCAGCCACACC (SEQ ID NO: 567).
  • a 3’UTR is an FI element as described in WO2017/060314, which is herein incorporated by reference in its entirety.
  • RNA Formats At least three distinct formats useful for RNA compositions (e.g., pharmaceutical compositions) have been developed, namely non-modified uridine containing mRNA (uRNA), nucleoside-modified mRNA (modRNA), and self-amplifying mRNA (saRNA). Each of these platforms displays unique features. In general, in all three formats, RNA is capped, contains open reading frames (ORFs) flanked by untranslated regions (UTR), and have a polyA-tail at the 3' end.
  • ORFs open reading frames flanked by untranslated regions
  • an ORF of an uRNA and modRNA vectors encode an antibody agent or portion thereof.
  • An saRNA has multiple ORFs.
  • the RNA described herein may have modified nucleosides.
  • the RNA comprises a modified nucleoside in place of at least one (e.g., every) uridine.
  • uracil describes one of the nucleobases that can occur in the nucleic acid of RNA.
  • the structure of uracil is: .
  • uridine describes one of the nucleosides that can occur in RNA.
  • the structure of uridine is: .
  • UTP uridine 5’-triphosphate
  • Pseudo-UTP pseudouridine 5’-triphosphate
  • Pseudouridine is one example of a modified nucleoside that is an isomer of uridine, where the uracil is attached to the pentose ring via a carbon-carbon bond instead of a nitrogen-carbon glycosidic bond.
  • Another exemplary modified nucleoside is N1-methyl-pseudouridine (m1 ⁇ ), which has the structure: .
  • N1-methyl-pseudo-UTP has the following structure: .
  • RNA comprises a modified nucleoside in place of at least one uridine. In some embodiments, RNA comprises a modified nucleoside in place of each uridine.
  • the modified nucleoside is independently selected from pseudouridine ( ⁇ ), N1- methyl-pseudouridine (m1 ⁇ ), and 5-methyl-uridine (m5U).
  • the modified nucleoside comprises pseudouridine ( ⁇ ). In some embodiments, the modified nucleoside comprises N1-methyl-pseudouridine (m1 ⁇ ). In some embodiments, the modified nucleoside comprises 5-methyl-uridine (m5U). In some embodiments, RNA may comprise more than one type of modified nucleoside, and the modified nucleosides are independently selected from pseudouridine ( ⁇ ), N1-methyl-pseudouridine (m1 ⁇ ), and 5-methyl-uridine (m5U). In some embodiments, the modified nucleosides comprise pseudouridine ( ⁇ ) and N1-methyl-pseudouridine (m1 ⁇ ).
  • the modified nucleosides comprise pseudouridine ( ⁇ ) and 5-methyl-uridine (m5U). In some embodiments, the modified nucleosides comprise N1-methyl-pseudouridine (m1 ⁇ ) and 5-methyl-uridine (m5U). In some embodiments, the modified nucleosides comprise pseudouridine ( ⁇ ), N1-methyl-pseudouridine (m1 ⁇ ), and 5-methyl-uridine (m5U).
  • the modified nucleoside replacing one or more, e.g., all, uridine in the RNA may be any one or more of 3-methyl-uridine (m3U), 5-methoxy-uridine (mo5U), 5-aza-uridine, 6-aza-uridine, 2-thio-5- aza-uridine, 2-thio-uridine (s2U), 4-thio-uridine (s4U), 4-thio-pseudouridine, 2-thio-pseudouridine, 5-hydroxy-uridine (ho5U), 5-aminoallyl-uridine, 5-halo-uridine (e.g., 5-iodo-uridine or 5-bromo-uridine), uridine 5-oxyacetic acid (cmo5U), uridine 5-oxyacetic acid methyl ester (mcmo5U), 5-carboxymethyl-uridine (cm5U), 1-carboxymethyl- pseudouridine, 5-carboxyhydroxymethyl-uridine (chm
  • the RNA comprises other modified nucleosides or comprises further modified nucleosides, e.g., modified cytidine.
  • modified cytidine in the RNA 5-methylcytidine is substituted partially or completely, preferably completely, for cytidine.
  • the RNA comprises 5- methylcytidine and one or more selected from pseudouridine ( ⁇ ), N1-methyl-pseudouridine (m1 ⁇ ), and 5-methyl- uridine (m5U).
  • the RNA comprises 5-methylcytidine and N1-methyl-pseudouridine (m1 ⁇ ).
  • the RNA comprises 5-methylcytidine in place of each cytidine and N1-methyl-pseudouridine (m1 ⁇ ) in place of each uridine.
  • the RNA is “replicon RNA” or simply a “replicon,” in particular “self-replicating RNA” or “self-amplifying RNA.”
  • the replicon or self-replicating RNA is derived from or comprises elements derived from a single-stranded (ss) RNA virus, in particular a positive-stranded ssRNA virus, such as an alphavirus. Alphaviruses are typical representatives of positive- stranded RNA viruses.
  • Alphaviruses replicate in the cytoplasm of infected cells (for review of the alphaviral life cycle see Jose et al., Future Microbiol., 2009, vol. 4, pp. 837–856, which is incorporated herein by reference in its entirety).
  • the total genome length of many alphaviruses typically ranges between 11,000 and 12,000 nucleotides, and the genomic RNA typically has a 5’-cap, and a 3’ poly(A) tail.
  • the genome of alphaviruses encodes non-structural proteins (involved in transcription, modification and replication of viral RNA and in protein modification) and structural proteins (forming the virus particle). There are typically two open reading frames (ORFs) in the genome.
  • the four non-structural proteins are typically encoded together by a first ORF beginning near the 5′ terminus of the genome, while alphavirus structural proteins are encoded together by a second ORF which is found downstream of the first ORF and extends near the 3’ terminus of the genome.
  • first ORF is larger than the second ORF, the ratio being roughly 2:1.
  • RNA RNA molecule that resembles eukaryotic messenger RNA
  • mRNA messenger RNA
  • the (+) stranded genomic RNA directly acts like a messenger RNA for the translation of the open reading frame encoding the non-structural poly- protein (nsP1234).
  • Alphavirus-derived vectors have been proposed for delivery of foreign genetic information into target cells or target organisms.
  • a first ORF encodes an alphavirus-derived RNA-dependent RNA polymerase (replicase), which upon translation mediates self-amplification of the RNA.
  • a second ORF encoding alphaviral structural proteins is replaced by an open reading frame encoding a malarial T cell peptide string construct described herein.
  • Alphavirus-based trans-replication systems rely on alphavirus nucleotide sequence elements on two separate nucleic acid molecules: one nucleic acid molecule encodes a viral replicase, and the other nucleic acid molecule is capable of being replicated by said replicase in trans (hence the designation trans-replication system).
  • Trans-replication requires the presence of both these nucleic acid molecules in a given host cell.
  • the nucleic acid molecule capable of being replicated by the replicase in trans must comprise certain alphaviral sequence elements to allow recognition and RNA synthesis by the alphaviral replicase.
  • a non-modified uridine platform may include, for example, one or more of intrinsic adjuvant effect, as well as good tolerability and safety.
  • modified uridine (e.g., pseudouridine) platform may include reduced adjuvant effect, blunted immune innate immune sensor activating capacity and thus good tolerability and safety.
  • RNA constructs optimized for example, for improved manufacturability, encapsulation, expression level (and/or timing), etc. Certain components are discussed below, and certain preferred embodiments are exemplified herein.
  • an “ERMA” construct is an “RNA construct,” and, for example, “ERMA 1” corresponds to “RNA Construct 1”, “ERMA 2” corresponds to “RNA construct 2,” etc. 3.
  • Codon Optimization and GC Enrichment refers to alteration of codons in a coding region of a nucleic acid molecule (e.g., a polyribonucleotide) to reflect the typical codon usage of a host organism (e.g., a subject receiving a nucleic acid molecule (e.g., a polyribonucleotide)) without preferably altering the amino acid sequence encoded by the nucleic acid molecule.
  • coding regions are codon-optimized for optimal expression in a subject to be treated using the RNA molecules described herein.
  • codon-optimization may be performed such that codons for which frequently occurring tRNAs are available are inserted in place of “rare codons.”
  • codon-optimization may include increasing guanosine/cytosine (G/C) content of a coding region of RNA described herein as compared to the G/C content of the corresponding coding sequence of a wild-type RNA, wherein the amino acid sequence encoded by the RNA is preferably not modified compared to the amino acid sequence.
  • G/C guanosine/cytosine
  • a coding sequence (also referred to as a “coding region”) is codon optimized for expression in the subject to whom a composition (e.g., a pharmaceutical composition) is to be administered (e.g., a human).
  • a composition e.g., a pharmaceutical composition
  • sequences in such a polynucleotide may differ from wild type sequences encoding the relevant antigen or fragment or epitope thereof, even when the amino acid sequence of the antigen or fragment or epitope thereof is wild type.
  • strategies for codon optimization for expression in a relevant subject e.g., a human
  • a relevant subject e.g., a human
  • Various species exhibit particular bias for certain codons of a particular amino acid.
  • codon bias differences in codon usage between organisms
  • codon bias often correlates with the efficiency of translation of messenger RNA (mRNA), which is in turn believed to be dependent on, among other things, the properties of the codons being translated and the availability of particular transfer RNA (tRNA) molecules.
  • mRNA messenger RNA
  • tRNA transfer RNA
  • the predominance of selected tRNAs in a cell may generally be a reflection of the codons used most frequently in peptide synthesis.
  • genes may be tailored for optimal gene expression in a given organism based on codon optimization.
  • Codon usage tables are available, for example, at the "Codon Usage Database” available at www.kazusa.orjp/codon/ and these tables may be adapted in a number of ways.
  • Computer algorithms for codon optimizing a particular sequence for expression in a particular subject or its cells are also available, such as Gene Forge (Aptagen; Jacobus, PA), are also available.
  • a polynucleotide (e.g., a polyribonucleotide) of the present disclosure is codon optimized, wherein the codons in the polynucleotide (e.g., the polyribonucleotide) are adapted to human codon usage (herein referred to as “human codon optimized polynucleotide”). Codons encoding the same amino acid occur at different frequencies in a subject, e.g., a human.
  • the coding sequence of a polynucleotide of the present disclosure is modified such that the frequency of the codons encoding the same amino acid corresponds to the naturally occurring frequency of that codon according to the human codon usage, e.g., as shown in Table 8.
  • the wild type coding sequence is preferably adapted in a way that the codon “GCC” is used with a frequency of 0.40, the codon “GCT” is used with a frequency of 0.28, the codon “GCA” is used with a frequency of 0.22 and the codon “GCG” is used with 30 a frequency of 0.10 etc. (see Table 8).
  • such a procedure (as exemplified for Ala) is applied for each amino acid encoded by the coding sequence of a polynucleotide to obtain sequences adapted to human codon usage.
  • Table 8 Human codon usage table with frequencies indicated for each amino acid
  • a coding sequence may be optimized using a multiparametric optimization strategy.
  • optimization parameters may include parameters that influence protein expression, which can be, for example, impacted on a transcription level, an mRNA level, and/or a translational level.
  • exemplary optimization parameters include, but are not limited to transcription-level parameters (including, e.g., GC content, consensus splice sites, cryptic splice sites, SD sequences, TATA boxes, termination signals, artificial recombination sites, and combinations thereof); mRNA-level parameters (including, e.g., RNA instability motifs, ribosomal entry sites, repetitive sequences, and combinations thereof); translation-level parameters (including, e.g., codon usage, premature poly(A) sites, ribosomal entry sites, secondary structures, and combinations thereof); or combinations thereof.
  • a coding sequence may be optimized by a GeneOptimizer algorithm as described in Fath et al.
  • a coding sequence may be optimized by Eurofins’ adaption and optimization algorithm “GENEius” as described in Eurofins’ Application Notes: Eurofins’ adaption and optimization software “GENEius” in comparison to other optimization algorithms, the entire content of which is incorporated by reference for the purposes described herein.
  • a coding sequence utilized in accordance with the present disclosure has G/C content that is increased compared to a wild type coding sequence for a malarial construct described herein, or a portion thereof.
  • guanosine/cytidine (G/C) content of a coding region is modified relative to a wild type coding sequence for a malarial construct described herein, but the amino acid sequence encoded by the polyribonucleotide not modified.
  • G/C guanosine/cytidine
  • GC enrichment may improve translation of a payload sequence.
  • sequences having an increased G (guanosine)/C (cytidine) content are more stable than sequences having an increased A (adenosine)/U (uridine) content.
  • codons which contain A and/or U nucleosides can be modified by substituting these codons by other codons, which code for the same amino acids but contain no A and/or U or contain a lower content of A and/or U nucleosides.
  • G/C content of a coding region of a polyribonucleotide described herein is increased by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, or even more compared to the G/C content of the coding region prior to codon optimization, e.g., of the wild type RNA.
  • G/C content of a coding region of a polyribonucleotide described herein is decreased by at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, or even more compared to the G/C content of the coding region prior to codon optimization, e.g., of the wild type RNA.
  • stability and translation efficiency of a polyribonucleotide may incorporate one or more elements established to contribute to stability and/or translation efficiency of the polyribonucleotide; exemplary such elements are described, for example, in PCT/EP2006/009448 incorporated herein by reference.
  • a polyribonucleotide may be modified within the coding region, i.e., the sequence encoding the expressed peptide or protein, without altering the sequence of the expressed peptide or protein, for example so as to increase the GC- content to increase mRNA stability and/or to perform a codon optimization and, thus, enhance translation in cells.
  • a polyribonucleotide encoding a Plasmodium T-cell string polypeptide construct as described herein has a nucleotide sequence provided in Table 9 or 10.
  • Exemplary Plasmodium T-cell string polypeptide constructs are also shown schematically in FIG.3.
  • Table 9 Exemplary DNA Sequences Encoding a Plasmodium T-cell String Polypeptide Construct 57 57/ Table 10: Exemplary RNA Sequences Encoding a Plasmodium T-cell string Polypeptide Construct
  • Linker Sequences coding for peptide linkers as described in Section IIC(iii) above.
  • the linker is selected from an amino acid sequence as defined in Table 7.
  • a linker has the amino acid sequence GGSGGGGSGG (SEQ ID NO: 404).
  • a linker has the amino acid sequence GGGS (SEQ ID NO: 411).
  • a linker has the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 408).
  • a linker has the amino acid sequence AGNRVRRSVG (SEQ ID NO: 412).
  • FI element The 3'-UTR sequence as described in Section IID(i) above. In some embodiments, FI comprises the nucleotide sequence of SEQ ID NO: 567.
  • A30L70 A poly(A)-tail as described in Section IID(i) above. In some embodiments, A30L70 comprises the nucleotide sequence of SEQ ID NO: 569.
  • a polyribonucleotide encoding a malarial T cell peptide string construct described herein has one of the following structures: cap-hAg-Kozak-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-FI-A30L70; cap-hAg-Kozak-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-FI-A30L70; cap-hAg-Kozak-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-FI- A30L70; cap-hAg-Kozak-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-Antigen-Linker-
  • the sequence encoding a malarial T cell peptide string construct described herein comprises a modified nucleoside replacing (partially or completely, preferably completely) uridine, wherein the modified nucleoside is selected from the group consisting of pseudouridine ( ⁇ ), N1-methyl-pseudouridine (m1 ⁇ ), and 5-methyl-uridine.
  • the sequence encoding a malarial T cell peptide string construct described herein is codon-optimized.
  • the G/C content of the sequence encoding a malarial T cell peptide string construct described herein is increased compared to the wild type coding sequence.
  • the RNA (in particular, mRNA) described herein comprises: a 5’ UTR comprising the nucleotide sequence of SEQ ID NO: 563, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 562; a 3’ UTR comprising the nucleotide sequence of SEQ ID NO: 567, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 566; and a poly-A sequence comprising the nucleotide sequence of SEQ ID NO: 569.
  • the RNA (in particular, mRNA) described herein comprises: m27,3’-OGppp(m12’-O) ApG as capping structure at the 5'-end of the mRNA; a 5’ UTR comprising the nucleotide sequence of SEQ ID NO: 563, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 563; a 3’ UTR comprising the nucleotide sequence of SEQ ID NO: 567, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 567; and a poly-A sequence comprising the nucleotide sequence of SEQ ID NO: 569.
  • the RNA is unmodified. In some embodiments, the RNA is modified. In some embodiments, the RNA comprises N1-methyl-pseudouridine (m1 ⁇ ) in place of at least one uridine (e.g., in place of each uridine).
  • m1 ⁇ N1-methyl-pseudouridine
  • the RNA (in particular, mRNA) described herein comprises: m27,3’-OGppp(m12’-O) ApG as capping structure at the 5'-end of the mRNA; a 5’ UTR comprising the nucleotide sequence of SEQ ID NO: 563, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 563; a 3’ UTR comprising the nucleotide sequence of SEQ ID NO: 567, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 567; a poly-A sequence comprising the nucleotide sequence of SEQ ID NO: 569; and N1-methyl-pseudouridine (m1 ⁇
  • a malarial T cell peptide string construct described herein includes one or more T-cell antigens from proteins of Plasmodium falciparum.
  • F. Certain Exemplary Polyribonucleotides Encoding Plasmodium CSP Polypeptide Constructs [0676] In some embodiments, a polyribonucleotide encoding a Plasmodium CSP polypeptide construct as described herein has a nucleotide sequence provided in Table 11. Table 11: Exemplary RNA Sequences Encoding a Plasmodium CSP Polypeptide Construct
  • hAg-Kozak 5'-UTR sequence as described in Section IID(i) above.
  • hAg- Kozak/5’UTR comprises the nucleotide sequence of SEQ ID NO: 565.
  • hAg-Kozak comprises the nucleotide sequence of SEQ ID NO: 565.
  • sec Sequences encoding a secretory signal as described in Section IIC(i) above.
  • the secretory signal is from Plasmodium falciparum, preferably Plasmodium falciparum isolate 3D7, and referred to herein as Pfsec and has the sequence as defined in Table 5 (SEQ ID NO: 332).
  • the secretory signal is heterologous and selected from the amino acid sequences as defined in Table 5.
  • the secretory signal is HSV1-gD and referred to herein as HSV-1gDsec and has the sequence as defined in Table 5 (SEQ ID NO: 314, SEQ ID NO: 317, SEQ ID NO: 320).
  • Antigen Sequences encoding a Plasmodium CSP polypeptide construct, including regions from Plasmodium falciparum, preferably Plasmodium falciparum isolate 3D7, as described herein.
  • TMD Sequences encoding a transmembrane region as described in Section IIC(ii) above.
  • the TMD is a glycosylphosphatidylinositol (GPI) anchor region from Plasmodium CSP, preferably from Plasmodium falciparum isolate 3D7, and referred to herein as PfTMD and has the amino acid sequence of SEQ ID NO: 385.
  • the TMD is heterologous.
  • the TMD is from HSV1-gD and referred to herein as HSV-1TMD and has the amino acid sequence of SEQ ID NO: 379.
  • Linker Sequences coding for peptide linkers as described in Section IIC(iii) above.
  • FI element The 3'-UTR sequence as described in Section IID(i) above. In some embodiments, FI comprises the nucleotide sequence of SEQ ID NO: 567.
  • A30L70 A poly(A)-tail sequence as described in Section IID(i) above. In some embodiments, A30L70 comprises the nucleotide sequence of SEQ ID NO: 569.
  • a polyribonucleotide encoding a Plasmodium CSP polypeptide construct described herein has one of the following structures: cap-hAg-Kozak-Antigen-FI-A30L70; cap-hAg-Kozak-sec-Antigen-FI-A30L70; cap-hAg-Kozak-Antigen-TMD-FI-A30L70; or cap-hAg-Kozak-sec-Antigen-TMD-FI-A30L70.
  • the Antigen comprises a full-length CSP construct as defined above, a CSP construct with noncontiguous minor repeat regions as defined above, a N-terminal region deletion CSP construct as defined above, a N-terminal region and major region deleted CSP construct as defined above, a N-terminal domain deleted CSP construct as defined above, a N-terminal domain and major repeat region deleted CSP construct as defined above, a N-terminal domain and major repeat region deleted CSP construct with modified junction region variants or portions as defined above, a major repeat region portion and C-terminal region containing CSP constructs as defined above, an N-terminal and C-terminal deleted CSP construct with noncontiguous minor repeat regions as defined above, a 6NANP CSP construct as defined above, an 18 NANP CSP construct as defined above, a T-Cell C- term CSP construct as defined above, or a CSP construct set out in the Additional Select Exemplary CSP construct list.
  • a polyribonucleotide described herein has one of the following structures: cap-hAg-Kozak-Antigen-FI-A30L70; cap-hAg-Kozak-sec-Antigen-FI-A30L70; cap-hAg-Kozak-Antigen-TMD-FI-A30L70; cap-hAg-Kozak-sec-Antigen-TMD-FI-A30L70; cap-hAg-Kozak-Pfsec-Antigen-FI-A30L70; cap-hAg-Kozak-Antigen-PfTMD-FI-A30L70; cap-hAg-Kozak-Pfsec-Antigen-PfTMD-FI-A30L70; cap-hAg-Kozak-HSV-1gDsec-Antigen-FI-A30L70; cap-hAg-Kozak-Antigen-HSV-1TMD-FI-A30L70; cap-hAg-Kozak-Antigen-
  • the different elements may be linked by one or more linkers, e.g., a linker selected from an amino acid sequence as defined in Table 7.
  • a linker has the amino acid sequence GGSGGGGSGG (SEQ ID NO: 404).
  • a linker has the amino acid sequence GGGS.
  • a linker has the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO: 408).
  • a linker has the amino acid sequence AGNRVRRSVG (SEQ ID NO: 412).
  • the sequence encoding a Plasmodium CSP polypeptide construct described herein comprises a modified nucleoside replacing (partially or completely, preferably completely) uridine, wherein the modified nucleoside is selected from the group consisting of pseudouridine ( ⁇ ), N1-methyl-pseudouridine (m1 ⁇ ), and 5-methyl-uridine.
  • the sequence encoding a Plasmodium CSP polypeptide construct described herein is codon-optimized.
  • the G/C content of the sequence encoding a Plasmodium CSP polypeptide construct described herein is increased compared to the wild type coding sequence.
  • the RNA (in particular, mRNA) described herein comprises: a 5’ UTR comprising the nucleotide sequence of SEQ ID NO: 565, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 565; a 3’ UTR comprising the nucleotide sequence of SEQ ID NO: 567, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 567; and a poly-A sequence comprising the nucleotide sequence of SEQ ID NO: 569.
  • the RNA (in particular, mRNA) described herein comprises: m27,3’-OGppp(m12’-O) ApG as capping structure at the 5'-end of the mRNA; a 5’ UTR comprising the nucleotide sequence of SEQ ID NO: 565, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 565; a 3’ UTR comprising the nucleotide sequence of SEQ ID NO: 567, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 567; and a poly-A sequence comprising the nucleotide sequence of SEQ ID NO: 569.
  • the RNA is unmodified. In some embodiments, the RNA is modified. In some embodiments, the RNA comprises N1-methyl-pseudouridine (m1 ⁇ ) in place of at least one uridine (e.g., in place of each uridine).
  • m1 ⁇ N1-methyl-pseudouridine
  • the RNA (in particular, mRNA) described herein comprises: m27,3’-OGppp(m12’-O) ApG as capping structure at the 5'-end of the mRNA; a 5’ UTR comprising the nucleotide sequence of SEQ ID NO: 565, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 565; a 3’ UTR comprising the nucleotide sequence of SEQ ID NO: 567, or a nucleotide sequence having at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, or 80% identity to the nucleotide sequence of SEQ ID NO: 567; and a poly-A sequence comprising the nucleotide sequence of SEQ ID NO: 569; and N1-methyl-pseudouridine (m1
  • a Plasmodium CSP polypeptide construct described herein includes one or more malarial polypeptides or portions thereof from Plasmodium falciparum.
  • a Plasmodium CSP polypeptide construct described herein includes one or more regions or portions thereof derived from a Plasmodium falciparum CSP protein, an immunogenic variant thereof, or an immunogenic fragment of the Plasmodium falciparum CSP protein or the immunogenic variant thereof.
  • the RNA used in the present disclosure encodes an amino acid sequence comprising an Plasmodium falciparum CSP protein, an immunogenic variant thereof, or an immunogenic fragment of the Plasmodium falciparum CSP protein or the immunogenic variant thereof.
  • G. Certain Exemplary Combination of Plasmodium Polypeptide Constructs [0698] The present disclosure provides combinations of polyribonucleotides that can be used to express two or more Plasmodium polypeptide constructs.
  • a combination as described herein comprises two or more polyribonucleotides that encode one or more Plasmodium T-cell string polypeptide constructs as described herein and one or more polyribonucleotides that encode one or more Plasmodium CSP polypeptide constructs as described herein. In some embodiments, a combination as described herein comprises two or more polyribonucleotides that encode a Plasmodium T-cell string polypeptide construct as described herein and a Plasmodium CSP polypeptide construct as described herein.
  • a combination as described herein comprises one or two polyribonucleotides that encode a Plasmodium T-cell string polypeptide construct as described herein and one Plasmodium CSP polypeptide construct as described herein.
  • Certain exemplary combinations of Plasmodium T-cell string polypeptide constructs and Plasmodium CSP polypeptide constructs expressed by the one or more polyribonucleotides are included in Table 12 below.
  • combinations of polyribonucleotides can be used to express two Plasmodium polypeptide constructs, wherein the two Plasmodium polypeptide constructs expressed from the combination comprise amino acid sequences listed in the third and fourth columns of Table 12 below or amino acid sequences with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the amino acid sequences listed in the third and fourth columns of Table 12 below.
  • Table 12 Certain Exemplary Combinations
  • the present disclosure also provides combinations comprising three or more polyribonucleotides that encode a first Plasmodium T-cell string polypeptide construct as described herein, a second Plasmodium T-cell string polypeptide construct as described herein, and a Plasmodium CSP polypeptide construct as described herein.
  • Certain exemplary combinations of Plasmodium T-cell string polypeptide constructs and Plasmodium CSP polypeptide constructs expressed by a combination of the three or more polyribonucleotides are included in Table 13 below.
  • combinations of polyribonucleotides can be used to express three Plasmodium polypeptide constructs, wherein the three Plasmodium polypeptide constructs expressed from the combination comprise amino acid sequences listed in the fourth, fifth and sixth columns of Table 13 below or amino acid sequences with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the amino acid sequences listed in the fourth, fifth and sixth columns of Table 13 below.
  • Table 13 Certain Exemplary Combinations
  • a combination as described herein comprises a first polyribonucleotide encoding a Plasmodium T-cell string polypeptide construct and a second polyribonucleotide encoding a Plasmodium CSP polypeptide construct.
  • a first polyribonucleotide comprises the antigens included in Mas3a and a second polyribonucleotide comprises Plasmodium full-length CSP polypeptide construct including the Plasmodium CSP secretory signal and the Plasmodium CSP GPI anchor region.
  • the combination further comprises a third polyribonucleotide encoding a Plasmodium T-cell string polypeptide construct that comprises the antigens included in Mas4f.
  • a combination as described herein comprises a first polyribonucleotide encoding a Plasmodium T-cell string polypeptide construct and a second polyribonucleotide encoding a Plasmodium CSP polypeptide construct.
  • a Plasmodium T-cell string polypeptide construct comprises: (i) an antigenic Plasmodium CSP polypeptide fragment, (ii) an antigenic Plasmodium TRAP polypeptide fragment, (iii) an antigenic Plasmodium UIS3 polypeptide fragment, (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment, and (v) an antigenic Plasmodium LSAP2 polypeptide fragment.
  • a Plasmodium T-cell string polypeptide construct comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 203.
  • a Plasmodium CSP polypeptide construct comprises (i) a secretory signal, (ii) a Plasmodium CSP N-terminal region, (iii) a Plasmodium CSP N-terminal end region, (iv) a Plasmodium CSP junction region, (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (vi) a Plasmodium CSP major repeat region, (vii) a Plasmodium CSP C- terminal region, and (viii) a transmembrane region.
  • a Plasmodium CSP polypeptide construct comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 33.
  • a Plasmodium CSP polypeptide construct comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 33.
  • a combination further comprises a third polyribonucleotide that encodes a second Plasmodium T-cell string polypeptide construct.
  • a second Plasmodium T-cell string polypeptide construct comprises (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment, (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment, (iii) an antigenic Plasmodium LISP-2 polypeptide fragment, and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a second Plasmodium T-cell string polypeptide construct comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 209.
  • a combination as described herein comprises a first polyribonucleotide encoding a Plasmodium T-cell string polypeptide construct and a second polyribonucleotide encoding a Plasmodium CSP polypeptide construct.
  • a Plasmodium T-cell string polypeptide construct comprises (i) an antigenic Plasmodium CSP polypeptide fragment, (ii) an antigenic Plasmodium TRAP polypeptide fragment, (iii) an antigenic Plasmodium UIS3 polypeptide fragment, (iv) an antigenic Plasmodium ETRAMP10.3 polypeptide fragment, and (v) an antigenic Plasmodium LSAP2 polypeptide fragment.
  • a Plasmodium T-cell string polypeptide construct comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 203.
  • a Plasmodium CSP polypeptide construct comprises (i) a secretory signal, (ii) a Plasmodium CSP N-terminal end region, (iii) a Plasmodium CSP junction region, (iv) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (v) a Plasmodium CSP C-terminal region, (vi) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region, (vii) a linker, and (viii) a transmembrane region, and wherein the Plasmodium CSP polypeptide construct does not comprise any of (a) a Plasmodium CSP N-terminal region or portion thereof, and (b) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a Plasmodium CSP polypeptide construct comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 81.
  • a combination further comprises a third polyribonucleotide that encodes a second Plasmodium T-cell string polypeptide construct.
  • a second Plasmodium T-cell string polypeptide construct comprises (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment, (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment, (iii) an antigenic Plasmodium LISP-2 polypeptide fragment, and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a second Plasmodium T-cell string polypeptide construct comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 209.
  • a combination as described herein comprises a first polyribonucleotide encoding a Plasmodium T-cell string polypeptide construct and a second polyribonucleotide encoding a Plasmodium CSP polypeptide construct.
  • a Plasmodium T-cell string polypeptide construct comprises (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment, (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment, (iii) an antigenic Plasmodium LISP-2 polypeptide fragment, and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a Plasmodium T-cell string polypeptide construct comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 209.
  • a Plasmodium CSP polypeptide construct comprises (i) a secretory signal, (ii) a Plasmodium CSP N-terminal region, (iii) a Plasmodium CSP N-terminal end region, (iv) a Plasmodium CSP junction region, (v) three repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (vi) a Plasmodium CSP major repeat region, (vii) a Plasmodium CSP C-terminal region, and (viii) a transmembrane region.
  • a Plasmodium CSP polypeptide construct comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 33.
  • a Plasmodium CSP polypeptide construct comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 33.
  • a combination as described herein comprises a first polyribonucleotide encoding a Plasmodium T-cell string polypeptide construct and a second polyribonucleotide encoding a Plasmodium CSP polypeptide construct.
  • a Plasmodium T-cell string polypeptide construct comprises (i) an antigenic Plasmodium LSA-1(a) polypeptide fragment, (ii) an antigenic Plasmodium LSA-1(b) polypeptide fragment, (iii) an antigenic Plasmodium LISP-2 polypeptide fragment, and (iv) an antigenic Plasmodium LISP-1 polypeptide fragment.
  • a Plasmodium T-cell string polypeptide construct comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 209.
  • a Plasmodium CSP polypeptide construct comprises (i) a secretory signal, (ii) a Plasmodium CSP N-terminal end region, (iii) a Plasmodium CSP junction region, (iv) nine repeats of the amino acid sequence of NANPNVDP (SEQ ID NO: 223), (v) a Plasmodium CSP C-terminal region, (vi) a serine-valine sequence immediately following the Plasmodium CSP C-terminal region, (vii) a linker, and (viii) a transmembrane region, and wherein the Plasmodium CSP polypeptide construct does not comprise any of (a) a Plasmodium CSP N-terminal region or portion thereof, and (b) an amino acid sequence of NPNA (SEQ ID NO: 228).
  • a Plasmodium CSP polypeptide construct comprises or consists of an amino acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to an amino acid sequence according to SEQ ID NO: 81.
  • H. Certain Exemplary Combination of Polyribonucleotide Constructs [0707] As discussed throughout, the present disclosure provides combinations of polyribonucleotides that can be used to express one or more Plasmodium polypeptide constructs.
  • a combination as described herein comprises one or more polyribonucleotides that encode one or more Plasmodium T-cell string polypeptide constructs as described herein and one or more polyribonucleotides that encode one or more Plasmodium CSP polypeptide constructs as described herein.
  • a combination as described herein comprises a first polyribonucleotide that encodes a Plasmodium T-cell string polypeptide construct as described herein and a second polyribonucleotide that encodes a Plasmodium CSP polypeptide construct as described herein.
  • two polyribonucleotides in a combination can have a ribonucleic acid sequence as listed in the third and fourth columns of Table 12 above or ribonucleic acid sequences with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the ribonucleic acid sequences listed in the third and fourth columns of Table 12 above.
  • the present disclosure also provides combinations a first polyribonucleotide that encodes a first Plasmodium T-cell string polypeptide construct as described herein, a second polyribonucleotide that encodes a second Plasmodium T-cell string polypeptide construct as described herein, and a third polyribonucleotide that encodes a Plasmodium CSP polypeptide construct as described herein.
  • Certain exemplary combinations of Plasmodium T-cell string polypeptide constructs and Plasmodium CSP polypeptide constructs expressed by a combination of the three or more polyribonucleotides are included in Table 13 above.
  • three polyribonucleotides in a combination can have a ribonucleic acid sequences as listed in the fourth, fifth, and sixth columns of Table 13 above or ribonucleic acid sequences with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity with the ribonucleic acid sequences listed in the fourth, fifth, and sixth columns of Table 13 above.
  • a combination as described herein comprises a first polyribonucleotide encoding a Plasmodium T-cell string polypeptide construct and a second polyribonucleotide encoding a Plasmodium CSP polypeptide construct.
  • a first polyribonucleotide comprises a ribonucleic acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a ribonucleic acid sequence according to SEQ ID NO: 204.
  • a first polyribonucleotide comprises a ribonucleic acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a ribonucleic acid sequence according to SEQ ID NOs: 34, 145, 147, 149, 151, 153, 155, and 157.
  • a first polyribonucleotide comprises a ribonucleic acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a ribonucleic acid sequence according to SEQ ID NO: 147.
  • a combination as described herein further comprises a third polyribonucleotide encoding a second Plasmodium T-cell string polypeptide construct.
  • a third polyribonucleotide comprises a ribonucleic acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a ribonucleic acid sequence according to SEQ ID NO: 210.
  • a combination as described herein comprises a first polyribonucleotide encoding a Plasmodium T-cell string polypeptide construct and a second polyribonucleotide encoding a Plasmodium CSP polypeptide construct.
  • a first polyribonucleotide comprises a ribonucleic acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a ribonucleic acid sequence according to SEQ ID NO: 204.
  • a first polyribonucleotide comprises a ribonucleic acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a ribonucleic acid sequence according to SEQ ID NO: 82.
  • a combination as described herein further comprises a third polyribonucleotide encoding a second Plasmodium T-cell string polypeptide construct.
  • a third polyribonucleotide comprises a ribonucleic acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a ribonucleic acid sequence according to SEQ ID NO: 210.
  • a combination as described herein comprises a first polyribonucleotide encoding a Plasmodium T-cell string polypeptide construct and a second polyribonucleotide encoding a Plasmodium CSP polypeptide construct.
  • a first polyribonucleotide comprises a ribonucleic acid sequence with at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a ribonucleic acid sequence according to SEQ ID NO: 210.

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Abstract

La présente divulgation concerne des compositions (par exemple, des compositions pharmaceutiques) pour l'administration d'antigènes protéiques de Plasmodium et des technologies associées (par exemple, des composants correspondants et/ou des méthodes associées). Entre autres, la présente divulgation concerne des combinaisons comprenant une première composition pharmaceutique contenant un premier polyribonucléotide et une seconde composition pharmaceutique contenant un second polyribonucléotide. Dans certains modes de réalisation, un premier polyribonucléotide code un premier polypeptide qui comprend un ou plusieurs antigènes de lymphocytes T de Plasmodium. Dans certains modes de réalisation, un second polyribonucléotide code un second polypeptide qui comprend un ou plusieurs polypeptides de Plasmodium (par exemple, CSP) ou des parties antigéniques de celui-ci.
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Family Cites Families (56)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5965542A (en) 1997-03-18 1999-10-12 Inex Pharmaceuticals Corp. Use of temperature to control the size of cationic liposome/plasmid DNA complexes
JP2002502831A (ja) 1998-02-03 2002-01-29 イネックス ファーマシューティカルズ コーポレイション 癌の治療に用いる血清に対して安定なプラスミド脂質粒子の全身供与
US6410328B1 (en) 1998-02-03 2002-06-25 Protiva Biotherapeutics Inc. Sensitizing cells to compounds using lipid-mediated gene and compound delivery
US6211140B1 (en) 1999-07-26 2001-04-03 The Procter & Gamble Company Cationic charge boosting systems
ATE291925T1 (de) 2001-06-05 2005-04-15 Curevac Gmbh Stabilisierte mrna mit erhöhtem g/c-gehalt und optimierter codon usage für die gentherapie
EP1519714B1 (fr) 2002-06-28 2010-10-20 Protiva Biotherapeutics Inc. Appareil liposomal et procedes de fabrication
AU2004257373B2 (en) 2003-07-16 2011-03-24 Arbutus Biopharma Corporation Lipid encapsulated interfering RNA
US6927663B2 (en) 2003-07-23 2005-08-09 Cardiac Pacemakers, Inc. Flyback transformer wire attach method to printed circuit board
CN1882693B (zh) 2003-09-15 2012-08-15 普洛体维生物治疗公司 聚乙二醇修饰的脂质化合物及其应用
CA2569664C (fr) 2004-06-07 2013-07-16 Protiva Biotherapeutics, Inc. Arn interferant encapsule dans des lipides
US7745651B2 (en) 2004-06-07 2010-06-29 Protiva Biotherapeutics, Inc. Cationic lipids and methods of use
JP5639338B2 (ja) 2005-07-27 2014-12-10 プロチバ バイオセラピューティクス インコーポレイティッド リポソームの製造システムおよび製造方法
EP3100718B1 (fr) 2008-01-02 2019-11-27 Arbutus Biopharma Corporation Compositions améliorées et procédés d'administration d'acides nucléiques
EP2279254B1 (fr) 2008-04-15 2017-07-05 Protiva Biotherapeutics Inc. Nouvelles formulations lipidiques pour l'administration d'acides nucléiques
WO2010053572A2 (fr) 2008-11-07 2010-05-14 Massachusetts Institute Of Technology Lipidoïdes aminoalcool et leurs utilisations
EP3699172A3 (fr) 2008-11-10 2020-11-18 Arbutus Biopharma Corporation Nouveaux lipides et compositions d'administration d'agents thérapeutiques
ES2613498T3 (es) 2009-07-01 2017-05-24 Protiva Biotherapeutics Inc. Nuevas formulaciones de lípidos para el suministro de agentes terapéuticos a tumores sólidos
US8569256B2 (en) 2009-07-01 2013-10-29 Protiva Biotherapeutics, Inc. Cationic lipids and methods for the delivery of therapeutic agents
WO2011066651A1 (fr) 2009-12-01 2011-06-09 Protiva Biotherapeutics, Inc. Préparations de snalp contenant des antioxydants
CA3044884A1 (fr) 2009-12-07 2011-06-16 Arbutus Biopharma Corporation Compositions utilisees pour l'administration d'acides nucleiques
US20130017223A1 (en) 2009-12-18 2013-01-17 The University Of British Columbia Methods and compositions for delivery of nucleic acids
US20130123338A1 (en) 2010-05-12 2013-05-16 Protiva Biotherapeutics, Inc. Novel cationic lipids and methods of use thereof
EP4481047A3 (fr) 2010-06-03 2025-04-16 Alnylam Pharmaceuticals, Inc. Lipides biodégradables pour l'administration d'agents actifs
WO2012016184A2 (fr) 2010-07-30 2012-02-02 Alnylam Pharmaceuticals, Inc. Procédés et compositions pour la délivrance d'agents actifs
US8466122B2 (en) 2010-09-17 2013-06-18 Protiva Biotherapeutics, Inc. Trialkyl cationic lipids and methods of use thereof
JP6293485B2 (ja) * 2010-09-27 2018-03-14 ザ トラスティーズ オブ ザ ユニバーシティ オブ ペンシルバニア コンセンサス抗原コンストラクトおよびそれらから作製されるワクチン、ならびにそれらを使用するマラリアの治療方法
US9999673B2 (en) 2011-01-11 2018-06-19 Alnylam Pharmaceuticals, Inc. PEGylated lipids and their use for drug delivery
US8691750B2 (en) 2011-05-17 2014-04-08 Axolabs Gmbh Lipids and compositions for intracellular delivery of biologically active compounds
LT2714071T (lt) * 2011-05-24 2019-10-10 Biontech Rna Pharmaceuticals Gmbh Individualizuotos vakcinos nuo vėžio
KR20220025112A (ko) 2011-06-08 2022-03-03 샤이어 휴먼 지네틱 테라피즈 인크. Mrna 전달을 위한 지질 나노입자 조성물 및 방법
WO2013016058A1 (fr) 2011-07-22 2013-01-31 Merck Sharp & Dohme Corp. Nouveaux lipides cationiques contenant du bis-azote pour administration d'oligonucléotide
JP6250543B2 (ja) 2011-09-27 2017-12-20 アルニラム・ファーマシューティカルズ・インコーポレーテッド ジ脂肪族置換ｐｅｇ化脂質
US8762704B2 (en) 2011-09-29 2014-06-24 Apple Inc. Customized content for electronic devices
WO2013086322A1 (fr) 2011-12-07 2013-06-13 Alnylam Pharmaceuticals, Inc. Lipides biodégradables ramifiés à terminaisons alkyle et cycloalkyle destinés à l'administration d'agents actifs
US20140308304A1 (en) 2011-12-07 2014-10-16 Alnylam Pharmaceuticals, Inc. Lipids for the delivery of active agents
DE21212055T1 (de) 2011-12-07 2022-08-04 Alnylam Pharmaceuticals, Inc. Biologisch abbaubare lipide zur freisetzung von wirkstoffen
ES2923757T3 (es) 2011-12-16 2022-09-30 Modernatx Inc Composiciones de ARNm modificado
WO2013143555A1 (fr) 2012-03-26 2013-10-03 Biontech Ag Formulation d'arn pour immunothérapie
US9415109B2 (en) 2012-07-06 2016-08-16 Alnylam Pharmaceuticals, Inc. Stable non-aggregating nucleic acid lipid particle formulations
ES3032935T3 (en) 2013-10-22 2025-07-29 Translate Bio Inc Lipid formulations for delivery of messenger rna
EP2923709A1 (fr) * 2014-03-28 2015-09-30 Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. Vaccin contre la malaria multi-composants-multi-étapes
CA2953341C (fr) 2014-06-25 2023-01-24 Acuitas Therapeutics Inc. Lipides et formulations de nanoparticules de lipides pour l'administration d'acides nucleiques
WO2016005004A1 (fr) 2014-07-11 2016-01-14 Biontech Rna Pharmaceuticals Gmbh Stabilisation de séquences d'adn codant pour une séquence poly (a)
HUE067372T2 (hu) 2015-06-29 2024-10-28 Acuitas Therapeutics Inc Lipidek és nanoszemcsés lipid formulázások nukleinsavak beadására
WO2017059902A1 (fr) 2015-10-07 2017-04-13 Biontech Rna Pharmaceuticals Gmbh Séquences utr 3' permettant la stabilisation d'arn
WO2017070624A1 (fr) * 2015-10-22 2017-04-27 Modernatx, Inc. Vaccins contre des maladies tropicales
CN108368028B (zh) 2015-10-28 2021-09-03 爱康泰生治疗公司 用于递送核酸的新型脂质和脂质纳米颗粒制剂
WO2018081480A1 (fr) 2016-10-26 2018-05-03 Acuitas Therapeutics, Inc. Formulations de nanoparticules lipidiques
EP3374504B1 (fr) 2015-11-09 2025-03-19 CureVac SE Molécules d'acide nucléique optimisées
SI3445850T1 (sl) 2016-04-22 2021-12-31 BioNTech SE Postopki za zagotavljanje enoverižne RNA
WO2017201454A1 (fr) * 2016-05-19 2017-11-23 Weiner, David Immunogènes synthétiques du paludisme, combinaisons de ceux-ci et leur utilisation pour prévenir et traiter les infections malariques
KR20190093816A (ko) 2016-10-26 2019-08-26 큐어백 아게 지질 나노입자 mRNA 백신
EP3357506A1 (fr) * 2017-02-02 2018-08-08 Institut Pasteur Plusieurs antigènes pré-érythrocytiques du paludisme et leur utilisation dans le déclenchement d'une réponse immunitaire protectrice chez un hôte
EP3796931A1 (fr) * 2018-05-23 2021-03-31 Institut Pasteur Antigènes pré-érythrocytaires du paludisme sous la forme de polypeptide de fusion et leur utilisation dans l'élicitation d'une réponse immunitaire protectrice chez un hôte
AU2019410737A1 (en) 2018-12-21 2021-06-10 CureVac SE RNA for malaria vaccines
IL319414A (en) * 2022-09-23 2025-05-01 BioNTech SE Compositions for delivering antigens at the liver level and related methods

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