Multabody constructs, compositions, and methods targeting malaria parasite proteins

Abstract

A fusion polypeptide comprises: (1) a malaria-associated antigen-binding moiety and (2) a nanocage monomer or subunit thereof, wherein the malaria-associated antigen-binding moiety comprises an antibody or antigen-binding fragment thereof comprising a heavy chain variable region (VH) comprising a CDR-H1, CDR-H2, and CDR-H3 having sequences which each differ by at most two amino acid residues from the CDR-H1, CDR-H2, and CDR-H3 sequences of a Pfs230 antibody, respectively; and / or a light chain variable region (VL or VK) comprising a CDR-L1, CDR-L2, and CDR-L3 having sequences which each differ by at most two amino acid residues from the CDR-L1, CDR-L2, and CDR-L3 sequences of a Pfs230 antibody, respectively.

Description

Attorney Docket No. RBT-014WOMULTABODY CONSTRUCTS, COMPOSITIONS, AND METHODS TARGETING MALARIA PARASITE PROTEINSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims benefit of and priority to U. S. Provisional Patent Application No. 63 / 759,880 filed February 18, 2025, the entire contents of which are incorporated by reference herein for all purposes.SEQUENCE LISTING

[0002] The present specification makes reference to a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML file, created on February 5, 2026, is named RBT-014WO_SL.xml and is 58,666 kilobytes in size.BACKGROUND

[0003] Malaria infections cause about 263 million clinical cases, and about 500,000 to 600,000 deaths worldwide annually. Malaria is a mosquito-borne infectious disease caused by protozoan parasites from the genus Plasmodium. Anopheles mosquitoes transmit malaria after being infected through a previous blood meal taken from an infected individual. There are four main types of malaria parasites which infect humans: Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium knowlesi, and Plasmodium ovale. Of those, falciparum malaria is the deadliest type. Many malaria parasites are now immune to the most common drugs used to treat the disease. According to the Malaria Eradication Research Agenda initiative, malaria eradication will be only achievable through effective vaccination. However, currently available malaria vaccine candidates have demonstrated only modest results in protecting against malaria. Thus, there is a need for improved therapeutics for treatment of malaria.SUMMARY

[0004] The present invention addresses this need with the provision of self-assembled polypeptide complexes which display IgG Fc fragments and malaria-associatedantigen-binders. Such self-assembled polypeptide complexes may be used to facilitate killingAttorney Docket No. RBT-014WOof pathogenic cells, including malaria parasites and / or cells (e.g., mammalian cells) infected therewith. In the provided self-assembled polypeptide complexes, the relative ratios, valency, and orientation of the binders and fragments are tunable, allowing modulation of the function and characteristics of the complexes. Also disclosed are related fusion proteins, complexes, compositions, and methods.

[0005] In one aspect, the disclosure provides fusion polypeptides comprising: (1) a malaria-associated antigen-binding moiety and (2) a nanocage monomer or subunit thereof, wherein the malaria-associated antigen-binding moiety comprises an antibody or antigen-binding fragment thereof comprising a heavy chain variable region (VH) comprising a CDR-H1, CDR-H2, and CDR-H3 having sequences which each differ by at most two amino acid residues from the CDR-H1, CDR-H2, and CDR-H3 sequences of a Pfs230 antibody, respectively; and / or a light chain variable region (VL or VK) comprising a CDR-L1, CDR-L2, and CDR-L3 having sequences which each differ by at most two amino acid residues from the CDR-L1, CDR-L2, and CDR-L3 sequences of a Pfs230 antibody, respectively.

[0006] In one aspect, the disclosure provides fusion polypeptides comprising: (1) a malaria-associated antigen-binding moiety and (2) a nanocage monomer or subunit thereof, wherein the malaria-associated antigen-binding moiety is capable of binding to an epitope of Pfs230. In some embodiments, the epitope comprises (a) amino acid residues E565, T566, T567, E568, S569, G570, D571, A573, V574, S575, E576, D577, Y579, K581, K610, E612, K653, E655, T656, K657, K698, A699, T700, V701, Y703, E724, and Y726 of SEQ ID NO: 50; or (b) amino acid residues D580, K581, Y582, A583, S584, V592, C593, D594, F595, D597, Q598, K600, P601, T602, E603, K607, V608, C611, E612, V632, L635, K716, andN719 of SEQ ID NO: 50. In some embodiments, the epitope of (b) further comprises an N585 amino acid residue.

[0007] In some embodiments, the malaria-associated antigen-binding moiety comprises an antibody or antigen-binding fragment thereof, such as an antibody or antigen-binding fragment thereof which comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL or VK).

[0008] In some embodiments, the VH comprises complementarity -determining regions (CDRs) CDR-H1, CDR-H2, and CDR-H3 having sequences which each differ by at most two amino acid residues from the sequences of SEQ ID NOs: 10, 11, and 12, respectively; and the VL or VK comprises CDRs CDR-L1, CDR-L2, and CDR-L3 having sequences which each differ by at most two amino acid residues from the sequences of SEQ ID NOs: 13, 14, and 15, respectively. In some embodiments, the CDR-H1, CDR-H2, and CDR-H3 have sequencesAttorney Docket No. RBT-014WOidentical to those of SEQ ID NOs: 10, 11, and 12, respectively; and the CDR-L1, CDR-L2, and CDR-L3 have sequences identical to those of SEQ ID NOs: 13, 14, andl5, respectively, except for one or two amino acid substitutions total across all six CDRs. In some embodiments, the CDR-H1, CDR-H2, and CDR-H3 have sequences identical to those of SEQ ID NOs: 10, 11, and 12, respectively; and the CDR-L1, CDR-L2, and CDR-L3 have sequences identical to those of SEQ ID NOs: 13, 14, 15, respectively. In some embodiments, the VH comprises a sequence having at least 85% identity to the sequence of SEQ ID NO: 8; and the VL or VK comprises a sequence having at least 85% identity to the sequence of SEQ ID NO: 9.

[0009] In some embodiments, the VH comprises CDRs CDR-H1, CDR-H2, and CDR-H3 having sequences which each differ by at most two amino acid residues from the sequences of SEQ ID NOs: 29, 30, and 31, respectively; and the Vi or VK comprises CDRs CDR-L1, CDR-L2, and CDR-L3 having sequences which each differ by at most two amino acid residues from the sequences of SEQ ID NOs: 32, 33, and 34, respectively. In some embodiments, the CDR-H1, CDR-H2, and CDR-H3 have sequences identical to those of SEQ ID NOs: 29, 30, and 31, respectively; andthe CDR-L1, CDR-L2, and CDR-L3 have sequences identical to those of SEQ ID NOs: 32, 33, and 34, respectively, except for one or two amino acid substitutions total across all six CDRs. In some embodiments, the CDR-H1, CDR-H2, and CDR-H3 have sequences identical to those of SEQ ID NOs: 29, 30, and 31, respectively; and the CDR-L1, CDR-L2, and CDR-L3have sequences identical to those of SEQ ID NOs: 32, 33, and 34, respectively. In some embodiments, the VH comprises a sequence having at least 85% identity to the sequence of SEQ ID NO: 27; and the VL or VK comprises a sequence having at least 85% identity to the sequence of SEQ ID NO: 28.

[0010] In some embodiments, the antibody or antigen-binding fragment thereof comprises or consists of a Fab fragment, such as a single-chain Fab fragment (scFab).

[0011] In some embodiments, the antibody or antigen-binding fragment thereof comprises or consists of an scFab which comprises a sequence having at least 80% identity to the sequence of SEQ ID NO: 51.

[0012] In some embodiments, the antibody or antigen-binding fragment thereof comprises or consists of an scFab which comprises a sequence having at least 80% identity to the sequence of SEQ ID NO: 53.

[0013] In some embodiments, the Pfs230 antibody is LMIV230-01.

[0014] In some embodiments, the Pfs230 antibody is RUPA-32.Attorney Docket No. RBT-014WO

[0015] In some embodiments, the nanocage monomer or subunit thereof is an apoferritin monomer or subunit thereof, such as a human apoferritin monomer or subunit thereof. In some embodiments, the apoferritin monomer is an apoferritin light chain.

[0016] In some embodiments, the malaria-associated antigen-binding moiety is linked via an amino acid linker to the nanocage monomer or subunit thereof, e.g., via the N-terminus of the nanocage monomer or subunit thereof.

[0017] In one aspect, the disclosure provides self-assembled polypeptide complexes comprising: (a) a plurality of malaria-associated antigen-binding fusion polypeptides, each fusion polypeptide being a fusion polypeptide as described herein; and (b) a plurality of Fc fusion polypeptides, each Fc fusion polypeptide comprising (1) an Fc polypeptide and (2) a nanocage monomer or subunit thereof.

[0018] In some embodiments, within each Fc fusion polypeptide, the Fc polypeptide is linked via an amino acid linker to the nanocage monomer or subunit thereof, e.g, via the N-terminus of the nanocage monomer or subunit thereof.

[0019] In one aspect, the disclosure provides self-assembled polypeptide complexes comprising: (a) a plurality of malaria-associated antigen-binding fusion polypeptides, each fusion polypeptide being a fusion polypeptide as disclosed herein; and (b) a plurality of Fc fusion polypeptides, each Fc fusion polypeptide comprising (1) an Fc polypeptide linked via an amino acid linker to the N-terminus of (2) a nanocage monomer subunit. In some embodiments, the nanocage monomer subunit is a C-half nanocage monomer.

[0020] In some embodiments, the C-half nanocage monomer is a C-half apoferritin and the nanocage monomers within the malaria-associated antigen-binding fusion polypeptide are each an apoferritin monomer or a subunit thereof. In some embodiments, the apoferritin monomer is a human apoferritin monomer. In some embodiments, the apoferritin monomer is an apoferritin light chain (e.g., human apoferritin light chain). In some embodiments, the selfassembled polypeptide complex of claim 33, which does not comprise any apoferritin heavy chains or subunits of apoferritin heavy chains. In some embodiments, the self-assembled polypeptide complex does not comprise or enclose iron.

[0021] In some embodiments, the self-assembled polypeptide complex comprises g a 2:1:1 ratio of a full-length human apoferritin; an Fc polypeptide fused to a first apoferritin monomer subunit; and a malaria-associated antigen-binding moiety fused to a second apoferritin monomer subunit.Attorney Docket No. RBT-014WO

[0022] In some embodiments, the self-assembled polypeptide complex comprises a 2:1:1 ratio of a full-length human apoferritin; an Fc polypeptide fused to C-half apoferritin; and a malaria-associated antigen-binding moiety fused to N-half apoferritin.

[0023] In some embodiments, the self-assembled polypeptide complex comprises a 2:1:1 ratio of a full-length human apoferritin; an Fc polypeptide fused to the N-terminus of C-half apoferritin; and a malaria-associated antigen-binding moiety fused to the N-terminus of N-half apoferritin.

[0024] In some embodiments, the Fc polypeptide is an IgGl Fc polypeptide.

[0025] In one aspect, provided are pharmaceutical compositions comprising a selfassembled polypeptide complex as disclosed herein and a pharmaceutically acceptable excipient.

[0026] In one aspect, provided are methods of treating, ameliorating, or preventing a disease or condition, the method comprising administering to a subject in need thereof a selfassembled polypeptide complex or pharmaceutical composition as disclosed herein.

[0027] In some embodiments, the subject is a mammal, e.g, a human.

[0028] In some embodiments, the disease or condition is malaria, e.g, malaria caused by an infection with P. falciparum.

[0029] In one aspect, the disclosure provides uses of a self-assembled polypeptide complexes or pharmaceutical compositions as disclosed herein to treat, ameliorate, or prevent a disease or condition (e.g., malaria, e.g., malaria caused by an infection with P. falciparum) in a subject in need thereof.

[0030] In one aspect, the disclosure provides methods comprising a step of contacting a cell with a self-assembled polypeptide complex as disclosed herein. In some embodiments, the cell is infected with a parasite, e.g., P. falciparum.

[0031] In some emobdiments, the step of contacting results in killing of the infected cell, such as an infected cell that expresses Pfs230. In some embodiments, the self-assembled polypeptide complex is capable of killing a parasite with an half-maximal inhibitory concentration (IC50) of no more than 10 nM, no more than 9 nM, no more than 8 nM, no more than 7 nM, no more than 6 nM, no more than 5 nM, no more than 4 nM, or no more than 3 nM.

[0032] In some embodiments, the step of contacting occurs inside a mammal (e.g., a human) having an infection.Attorney Docket No. RBT-014WOBRIEF DESCRIPTION OF THE DRAWINGS

[0033] FIG. 1A is a plot showing a luminescence signal (measured in relative light units (RLU)) generated in infected mosquitos by GFP:luc-expressing oocysts from a transgenic NF54-HGL P. falciparum line, wherein the infected mosquitos were treated with either (1) a multabody (“MB”) comprising fusion proteins comprising (a) an a-Pfs230 scFab derived from a LMIV230-01 parental antibody fused to N-half apoferritin (SEQ ID NO: 52) and (b) wild-type (WT) IgGl single chain Fc (scFc) polypeptide fused to C-half apoferritin (SEQ ID NO: 56), and (c) full-length apoferritin light chain (SEQ ID NO: 1) (“MB-WT”); (2) a full-length parental LMIV230-01 IgGl with WT Fc (“IgGl-WT”); (3) a MB comprising fusion proteins comprising (a) an a-Pfs230 scFab derived from LMIV230-01 fused to N-half apoferritin (SEQ ID NO: 52) and (b) an IgGl scFc polypeptide comprising “LLRAL” mutations fused to C-half apoferritin (SEQ ID NO: 49), and (c) full-length apoferritin light chain (SEQ ID NO: 1) (“MB-Fc mutant”); or (4) a full-length parental LMIV230-01 IgGl comprising LLRAL mutations (“IgGl-Fc mutant”). The luminescence intensity is shown as a function of the concentration of the MB or parental IgG (in nM). FIG. IB is a bar plot showing the half-maximal inhibitory concentration (IC50) of test molecules described in FIG.1A as a proxy for parasite-killing efficiency. FIG. 1C is a bar plot showing the IC50of test molecules similar to those described in FIG. 1A as a proxy for parasite-killing efficiency, except that the a-Pfs230 scFab is derived from RUPA-32, a Pfs230 antibody.

[0034] FIGS. 2A-2B are plots showing fluorescence intensity of P. falciparum gametocyte preparations treated with varying concentrations of (1) an MB comprising fusion proteins comprising (a) an a-Pfs230 scFab derived from a LMIV230-01 parental antibody (SEQ ID NO: 51) fused to N-half apoferritin (SEQ ID NO: 52) and (b) wild-type (WT) IgGl scFc polypeptide fused to C-half apoferritin (SEQ ID NO: 56), and (c) full-length apoferritin light chain (SEQ ID NO: 1) (“MB WT”; FIG. 2A); (2) a MB comprising fusion proteins comprising (a) an a-Pfs230 scFab derived from LMIV230-01 (SEQ ID NO: 51) fused to N-half apoferritin (SEQ ID NO: 52) and (b) an IgGl scFc polypeptide comprising “LLRAL” mutations fused to C-half apoferritin (SEQ ID NO: 49), and (c) full-length apoferritin light chain (SEQ ID NO: 1) (“MB mut”; FIG. 2A); (3) a full-length parental LMIV230-01 IgGl with WT Fc (“IgG WT”; FIG. 2B); or (4) a full-length parental LMIV230-01 IgGl comprising LLRAL mutations (“IgG mut”; FIG.2B). X-axis values correspond to exponent values, and the units are M (e.g., “-10” on the x-axis refers to 10-10M, “-9”on the x-axis refers to 10-9M).

[0035] FIGS. 3A-3B are plots showing fluorescence intensity of P. falciparum gametocyteAttorney Docket No. RBT-014WOpreparations treated with varying concentrations of (1) an MB comprising fusion proteins comprising (a) an a-Pfs230 scFab derived from a RUPA-32 parental antibody (SEQ ID NO: 53) fused to N-half apoferritin (SEQ ID NO: 54) and (b) wild-type (WT) IgGl scFc polypeptide fused to C-half apoferritin (SEQ ID NO: 56), and (c) full-length apoferritin light chain (SEQ ID NO: 1) (“MB WT”; FIG. 3A); (2) a MB comprising fusion proteins comprising (a) an a-Pfs230 scFab derived from RUPA-32 (SEQ ID NO: 53) fused to N-half apoferritin (SEQ ID NO: 54) and (b) an IgGl scFc polypeptide comprising “LLRAL” mutations fused to C-half apoferritin (SEQ ID NO: 49), and (c) full-length apoferritin light chain (SEQ ID NO: 1) (“MB mut”; FIG. 3A); (3) a full-length parental RUPA-32 IgGl with WT Fc (“IgG WT”; FIG. 3B); or (4) a full-length parental RUPA-32 IgGl comprising LLRAL mutations (“IgG mut”; FIG. 3B). X-axis values correspond to exponent values, and the units are M (e.g., “-10” on the x-axis refers to 10-10M, “-9”on the x-axis refers to 10-9M).DETAILED DESCRIPTION

[0036] Disclosed herein are fusion polypeptides each comprising (1) a nanocage monomer or subunit thereof and (2) an antibody fragment (such as a pathogen-associatedantigen-binder, e.g, a malaria-associated antigen binder) or an Fc polypeptide. The nanocage monomers or subunits thereof drive self-assembly of the fusion polypeptides into complexes which display one or more of the aforementioned malaria-associated antigen-binders and Fc polypeptides. Also disclosed are methods of treating, ameliorating, or preventing a disease or condition (e.g., an infectious disease or condition, such as malaria) by administering to a subject (e.g., a mammal, such as a human) a self-assembled polypeptide complex of the disclosure or a pharmaceutical composition comprising the same.Definitions

[0037] The terms “about” and “approximately,” when used herein in reference to a value, are used interchangeably and refer to a value that is similar to the referenced value. In general, those skilled in the art, familiar with the context, will appreciate the relevant degree of variation encompassed by “about” or “approximately” in that context. For example, in certain embodiments, the terms “about” and “approximately” may encompass a range of values that fall 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.Attorney Docket No. RBT-014WO

[0038] As used herein, the terms “alter,’’ “altered,’’ “decrease,’’ “decreased,’’ “increase,’’ “increased,” or “reduction,” “reduced,” (e.g., in reference to certain outcomes or effects) have meanings relative to a reference level. In certain embodiments, in the context of discussing mutations in an Fc chain or Fc polypeptide, the reference level is a level known or as determined with an IgG that does not contain the referenced mutation(s) in the Fc region.

[0039] As used herein, the term “antigen-binding fragment” of an antibody, refers to one or more fragments of an antibody that retain the ability to specifically bind to an antigen. Examples of binding fragments encompassed within the term “antigen binding fragment” of an antibody include a Fab fragment, a F(ab')2 fragment, a Fd fragment, a Fv fragment, a scFv fragment, a dAb fragment (Ward et al., (1989) Nature 341:544-546), and an isolated complementarity determining region (CDR). In certain embodiments, an “antigen binding fragment” comprises a heavy chain variable region and a light chain variable region. These antibody fragments can be obtained using conventional techniques known to those with skill in the art, and the fragments can be screened for utility in the same manner as are intact antibodies.

[0040] The term “apoferritin monomer,” is used herein to refer to a single chain of an apoferritin that, in the presence of other apoferritin chains, is capable of self-assembling into a polypeptide complex comprising a plurality of apoferritin chains, e.g, 24 or more apoferritin chains.

[0041] As used herein, the term “binding,” unless otherwise specified, refers to a non-covalent association between or among two or more entities. " Direct" binding involves physical contact between entities or moieties; indirect binding involves physical interaction by way of physical contact with one or more intermediate entities. Binding between two or more entities can typically be assessed in any of a variety of contexts, including where interacting entities or moieties are studied in isolation or in the context of more complex systems (e.g., while covalently or otherwise associated with a carrier entity and / or in a biological system or cell). As used herein, the phrases “non-binding” or “no binding,” or similar phrases, between two entities refers to 1) a lack of detectable binding or 2) binding below a set threshold that corresponds to no binding in an appropriate assay, e.g, an in vitro binding assay such as biolayer interferometry, surface plasmon resonance, a cell binding assay such as flow cytometry, or enzyme-linked immunosorbent assay (ELISA). For example, in certain embodiments, in an in vitro biolayer interferometry assay, a maximal association binding response of less than 0.1 nm after 180 seconds to a biosensor loaded withAttorney Docket No. RBT-014WO0.8 nm of target when the test article is present at a concentration of 20 nM is classified as “non-binding.”

[0042] The terms “ferritin” and “apoferritin” are used interchangeably herein and generally refer to a polypeptide (e.g., an apoferritin chain) that is capable of assembling into a ferritin complex (e.g., apoferritin complex) which typically comprises 24 protein subunits. In certain embodiments, the apoferritin is a human apoferritin, e.g, a human apoferritin light chain, e.g, a human apoferritin light chain having at least 85% sequence identity to SEQ ID NO: 1 or UniProt P02792. In certain embodiments, the apoferritin is a wild-type apoferritin. For example, the apoferritin may be a wild-type human apoferritin.

[0043] As used herein, the term “linker” is used to refer to an entity that connects two or more elements to form a multi-element agent. For example, those of ordinary skill in the art appreciate that a polypeptide (e.g., fusion polypeptide) whose structure includes two or more functional or organizational domains often includes a stretch of amino acids between such domains that links them to one another. In certain embodiments, a polypeptide comprising a linker element has an overall structure of the general form S1-L-S2, wherein SI and S2 may be the same or different and represent two domains associated with one another by the linker (L). In certain embodiments, the linker is an “amino acid linker,” that is, it comprises amino acid residues, e.g., an amino acid linker may comprise 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, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or more amino acid residues. In certain embodiments, a linker is characterized in that it tends not to adopt a rigid three-dimensional structure, but rather provides flexibility to the polypeptide.

[0044] The term “multispecific,” as used herein, refers to the characteristic of having at least two binding sites at which at least two different binding partners, e.g, an antigen or receptor (e.g., Fc receptor), can bind. For example, a polypeptide complex that comprises at least two Fab fragments, wherein each of the two Fab fragments is capable of binding (e.g., specifically binding) to a different antigen, is “multispecific.” As an additional example, a polypeptide complex that comprises an Fc fragment (which is capable of binding (e.g., specifically binding) to an Fc receptor) and a Fab fragment (which is capable of binding (e.g., specifically binding) to an antigen) is “multispecific.”

[0045] The term “multivalent,” as used herein, refers to the characteristic of having at least two binding sites at which a binding partner, e.g., an antigen or receptor (e.g., Fc receptor), can bind. The binding partners that can bind to at least two binding sites may be the same or different.Attorney Docket No. RBT-014WO

[0046] The term “nanocage monomer,” as used herein, refers to a single chain of a polypeptide that is capable of self-assembling with other nanocage monomers to form a self-assembled polypeptide complex comprising a plurality of nanocage monomers. In certain embodiments, the nanocage monomer is selected from monomers of ferritin, apoferritin, encapsulin, sulfur oxygenase reductase (SOR), lumazine synthase, pyruvate dehydrogenase, carboxysome, vault proteins, GroEL, heat shock protein, E2P coat protein, MS2 coat protein, fragments thereof, and variants thereof.

[0047] The term “polypeptide,” as used herein, generally has its art-recognized meaning of a polymer of at least three amino acids, e.g., linked to each other by peptide bonds. Those of ordinary skill in the art will appreciate that the term “polypeptide” is intended to be sufficiently general as to encompass not only polypeptides having a complete sequence recited herein, but also to encompass polypeptides that represent functional fragments (i.e., fragments retaining at least one activity) of such complete polypeptides. Moreover, those of ordinary skill in the art understand that protein sequences generally tolerate some substitution without destroying activity. Thus, any polypeptide that retains activity and shares at least about 30-40% overall sequence identity, often greater than about 50%, 60%, 70%, or 80%, and further usually including at least one region of much higher identity, often greater than 90% or even 95%, 96%, 97%, 98%, or 99% in one or more highly conserved regions, usually encompassing at least 3-4 and often up to 20 or more amino acids, with another polypeptide of the same class, is encompassed within the relevant term “polypeptide” as used herein. Polypeptides may contain L-amino acids, D-amino acids, or both and may contain any of a variety of amino acid modifications or analogs known in the art. Useful modifications include, e.g., terminal acetylation, amidation, methylation, glycosylation etc. In certain embodiments, proteins may comprise natural amino acids, non-natural amino acids, synthetic amino acids, and combinations thereof

[0048] As used herein, the term “specifically binds, “specifically binding,” “binds specifically,” or similar terms, means that a binding moiety (e.g., an antibody or an antigen-binding fragment thereof) forms a complex with an antigen that is relatively stable under physiologic conditions. Specific binding can be characterized by an equilibrium dissociation constant of 1×10-6M or less, 1×10-7M or less, 1×10-8M or less, or 1×10-9M or less (e.g., a smaller KD denotes a tighter binding). Methods for determining whether two molecules specifically bind are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, enzyme-linked immunosorbent assay, or biolayer interferometry measurements, etc. In certain embodiments, “specifically binds” and similarAttorney Docket No. RBT-014WOterms refers to a characteristic of the binding moiety in that the binding moiety is capable of binding to a target antigen but does is not capable of binding to other antigens such as distantly related family members of the antigen.

[0049] The term “self-assembled,” when used in reference to a macromolecular complex (e.g., a polypeptide complex), refers to the spontaneous formation of that complex when sufficient constituents of the complex (e.g., fusion polypeptides) to be formed are present. In certain embodiments, complexes self-assemble in physiological conditions, or in a buffer (e.g., a solution) that corresponds to physiological conditions.

[0050] As used herein, the term “subject” refers to an organism, typically a mammal (e.g., a human). In certain embodiments, a subject is suffering from or susceptible to a relevant disease, disorder or condition. In certain embodiments, a subject displays one or more symptoms or characteristics of a disease, disorder or condition. In certain embodiments, a subject is someone with one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition. In certain embodiments, a subject is a patient. In certain embodiments, a subject is a subject to whom diagnosis and / or therapy is and / or has been administered.

[0051] As used herein, the term “treatment” (also “treat” or “treating”) refers to any administration of a therapy that partially or completely alleviates, ameliorates, relieves, inhibits, delays onset of, reduces severity of, and / or reduces incidence of one or more symptoms, features, and / or causes of a particular disease, disorder, and / or condition. In certain embodiments, such treatment may be of a subject who does not exhibit signs of the relevant disease, disorder and / or condition and / or of a subject who exhibits only early signs of the disease, disorder, and / or condition. Alternatively, or additionally, such treatment may be of a subject who exhibits one or more established signs of the relevant disease, disorder and / or condition. In certain embodiments, treatment may be of a subject who has been diagnosed as suffering from the relevant disease, disorder, and / or condition. In certain embodiments, treatment may be of a subject known to have one or more susceptibility factors that are statistically correlated with increased risk of development of the relevant disease, disorder, and / or condition.

[0052] As used herein, the term “variant,” when used in reference to a polypeptide, e.g., an apoferritin monomer or subunit thereof, or an immunoglobulin molecule (such as an IgGl), refers to polypeptides having an amino acid sequence that differs from the sequence of the reference polypeptide by virtue of an insertion, deletion, modification and / or substitution of one or more amino acid residues within the sequence of the reference polypeptide but retainsAttorney Docket No. RBT-014WOat least some (if not all) biological activity of the reference polypeptide. A variant of a polypeptide typically has an amino acid sequence that has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the reference polypeptide. The term “variant” encompasses naturally-occurring variants (e.g., homologs and allelic variants) and non-naturally -occurring variants (e.g., mutants) of the polypeptide. Homologs of polypeptides differ in amino acid sequence from one species to another. Allelic variants of polypeptides differ in amino acid sequence from one individual to another within a species. Non-naturally -occurring variants include polypeptides that comprise a change in amino acid sequence, where the change in sequence is artificially introduced, e.g., the change is generated in the laboratory or other facility by human intervention (“hand of man”). Where particular polypeptides are disclosed herein, variants of such polypeptides are also contemplated and may be used in accordance with the fusion proteins, complexes, compositions, and method of the present disclosure.A. Fusion polypeptides

[0053] In many embodiments, fusion polypeptides compatible with compositions and methods disclosed herein generally comprise (1) a nanocage monomer (or subunit thereof) as described herein and (2) a cell-binder (e.g., a malaria-associated antigen binder) or an Fc polypeptide, which may be linked via a linker, such as a linker described herein.1. Nanocage monomers and subunits thereof

[0054] In certain embodiments, the nanocage monomer is an apoferritin monomer.

[0055] In certain embodiments, the apoferritin monomer is an apoferritin light chain. In certain embodiments, the apoferritin monomer does not include an apoferritin heavy chain or other apoferritin components capable of binding to iron or capable of ferroxidase activity.

[0056] In certain embodiments, each fusion polypeptide within the self-assembled polypeptide complex comprises an apoferritin light chain or a subunit of an apoferritin light chain. In these embodiments, the self-assembled polypeptide complex does not comprise any apoferritin heavy chains or subunits of apoferritin heavy chains. In such embodiments, the self-assembled polypeptide complex does not comprise and is not associated with Fe3+an iron ion normally carried by apoferritin complexes which comprise the apoferritin heavy chain.Attorney Docket No. RBT-014WO

[0057] In certain embodiments, the apoferritin monomer is a human apoferritin chain, e.g.. a human apoferritin light chain, e.g., a human apoferritin light chain having the sequence of at least residues 2-175 of SEQ ID NO: 1.

[0058] As used herein, a “full-length apoferritin light chain’’ refers to an apoferritin polypeptide that is capable of forming a four-helix bundle of two couples of a-parallel a-helices connected by a loop and further comprising a C-terminal a-helix. In certain embodiments, a full-length apoferritin light chain has an amino acid sequence which has at least 85%, at leat 87.5%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95% sequence, at least 96%, at least 97%, at least 98%, or at least 99% identity to the sequence corresponding to positions 2-175 of SEQ ID NO: 1. In certain embodiments, a full-length apoferritin light chain has an amino acid sequence corresponding to positions 2-175 of SEQ ID NO: 1. In certain embodiments, the full-length apoferritin light chain is a single contiguous apoferritin polypeptide.

[0059] A “subunit” of an apoferritin monomer refers to a portion of an apoferritin monomer that is capable of spontaneously associating with another, distinct subunit of an apoferritin monomer, so that the subunits together form an apoferritin monomer, which apoferritin monomer, in turn, is capable of self-assembling with other apoferritin monomers to form a polypeptide complex.

[0060] In certain embodiments, the apoferritin monomer subunit comprises approximately half of an apoferritin monomer. As used herein, the term “N-half apoferritin” refers to approximately half of an apoferritin chain, which half comprises the N-terminus of the apoferritin chain. As used herein, the term “C-half apoferritin” refers to approximately half an apoferritin chain, which half comprises the C-terminus of the apoferritin chain. The exact point at which an apoferritin chain may be divided to form the N-half apoferritin and the C-half apoferritin may vary depending on the embodiment. In the context of apoferritin monomer subunits based on human apoferritin light chain, for example, the halves may be divided at a point that corresponds to a position between about position 75 to about position 100 of SEQ ID NO: 1 (or a substantial portion thereof).

[0061] In certain embodiments, the halves are divided at a point that corresponds to a position between about position 85 to about position 95 of SEQ ID NO: 1. For example, in certain embodiments, an N-half apoferritin based on a human apoferritin light chain has an amino acid sequence corresponding to residues 1-90 of SEQ ID NO: 1 (or a substantial portion thereof, e.g, residues 2-90 of SEQ ID NO: 1), and a C-half apoferritin based on aAttorney Docket No. RBT-014WOhuman apoferritin light chain has an amino acid sequence corresponding to residues 91-175 of SEQ ID NO: 1 (or a substantial portion thereof).

[0062] Variants of the N-terminal or C-terminal portions of SEQ ID NO: 1 may also be used as the N-half apoferritin and C-half apoferritin, respectively. For example, in some embodiments, an N-half apoferritin has an amino acid sequence having at least 85%, at least 87.5%, 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% identical to that of SEQ ID NO: 2. In some embodiments, an N-half apoferritin has an amino acid sequence corresponding to residues 2-90 of SEQ ID NO: 2. In some embodiments, a C-half apoferritin has an amino acid sequence having at least 85%, at least 87.5%, 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% identical to that of SEQ ID NO: 3. In some embodiments, a C-half apoferritin has an amino acid sequence of SEQ ID NO: 3.2, Binding moieties

[0063] Binding moieties (e.g., malaria-associated antigen-binding moieties) typically comprise an antibody fragment.

[0064] In certain embodiments, the antibody fragment comprises or consists of a Fab. In certain embodiments, the antibody fragment comprises or consists of a single-chain Fab (scFab); for example, a fusion polypeptide comprising both the heavy and light chains of a Fab, optionally linked by a linker (e.g., amino acid linker as disclosed herein) is used.

[0065] In certain embodiments, the antibody fragment comprises a heavy chain variable region (e.g., a VH). In certain embodiments, the antibody fragment comprises a heavy chain variable domain (e.g., VH) and a light chain variable domain (e.g., a VL or VK). In certain embodiments, the antibody fragment comprises a Fab which comprises a heavy chain variable domain (e.g., VH) and a light chain variable domain (e.g., a VL or VK).

[0066] In certain embodiments, the antibody fragment does not comprise any domains from the Fc region, e.g., does not comprise any constant heavy (CH2 or CH3) domains. In certain embodiments, the antibody fragment is an antibody fragment of, or derived from, a fully human or humanized antibody. In certain embodiments, the antibody fragment is an antibody fragment of, or derived from, a chimeric antibody. The antibody from which the antibody fragment is obtained or derived can be of any of a variety of antibody classes, including, e.g., an IgGl antibody, an IgG2 antibody, or an IgG4 antibody.Attorney Docket No. RBT-014WO

[0067] In embodiments where multiple types of fusion polypeptides having antibody fragments are used, the antibody fragments in the various types of fusion polypeptides may be capable of binding to the same epitope on a given antigen (e.g., a malaria-associated antigen), capable of binding to epitopes that are distinct and non-overlapping on an antigen, or capable of binding to epitopes that are distinct but overlapping on the same antigen.A. Malaria-associated antigen-binding moieties

[0068] Malaria-associated antigen-binding moieties are generally capable of binding (e.g., specifically binding) to an antigen of a malaria protein (e.g., Pfs230). In certain embodiments, the malaria protein is expressed on the surface of malaria gametocytes or inside cells (e.g., mammalian cells, such as human cells) infected with the gametocytes.

[0069] In certain embodiments, the malaria-associated antigen-binding moiety is capable of binding (e.g., specifically binding) to Pfs230. In certain embodiments, the Pfs230 protein comprises an amino acid sequence of SEQ ID NO: 50 (UniProtKB ID: P68874.1) or a variant thereof having at least 85% sequence identity thereto.

[0070] In certain embodiments, the malaria-associated antigen-binding moiety comprises an antibody fragment comprising a heavy chain variable region (VH) and light chain variable region (VL or VK), wherein the CDRs of the VH and the VL or VK (CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3) have sequences which are similar to (e.g., each having at most one or two amino acids differing from) those of the heavy and light chain CDRs of a Pfs230 antibody (e.g., a human or humanized Pfs230 antibody).

[0071] In certain embodiments, the malaria-associated antigen-binding moiety is an antibody fragment comprising a heavy chain variable region (VH) and light chain variable region (VL or VK), wherein the CDRs of the VH and the VL or VK have sequences which are identical to those of the heavy and light chain of a Pfs230 antibody (e.g., a human or humanized Pfs230 antibody), except for one or two amino acid substitutions total across all six CDRs.

[0072] In certain embodiments, the malaria-associated antigen-binding moiety is an antibody fragment comprising a heavy chain variable region (VH) and light chain variable region (VL or VK), wherein the CDRs of the VH and the VL or VK are identical to those of the heavy and light chain CDRs of a Pfs230 antibody (e.g., a human or humanized Pfs230 antibody).

[0073] In certain embodiments, the malaria-associated antigen-binding moiety is an antibody fragment comprising heavy and light chain variable regions (e.g., VH and VL or VK) with sequences that have at least 85%, at least 87.5%, at least 90%, at least 91%, at leastAttorney Docket No. RBT-014WO92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity to those of the heavy and light chain variable regions of a Pfs230 antibody (e.g., a human or humanized Pfs230 antibody). In certain embodiments, themalaria-associated antigen-binding moiety is an antibody fragment comprising heavy and light chain variable regions (e.g., VH and VL or VK) with sequences identical to those of the heavy and light chain variable regions of a Pfs230 antibody (e.g., a human or humanized Pfs230 antibody).

[0074] In a non-limiting example, a Pfs230 antibody includes LMIV230-01 or an antigen-binding fragment thereof, those whose heavy chain variable region, light chain variable region, and CDR sequences are depicted in Tables 1A.

[0075] In another non-limiting example, a Pfs230 antibody includes RUPA-32 or an antigen-binding fragment thereof, those whose heavy chain variable region, light chain variable region, and CDR sequences are depicted in Tables IB.

[0076] Table 1A: Pfs230 Antibody (LMIV230-01) Sequences (VH / VL)Heavy chain variable region Light chain variable region QVQLVQSGAEVKKPGSSVKVSCKVSGGT DIVMTQSPASLALSLGERATLYCRASH FNTYAIIWVRQAPGQGLEWMGAIIPFQDR SISSWLAWYQQKPGKAPKLLIYKASTL GQYAQKFQGRVTITADKSTSTAYMELSSL ESGVPSRFSGSGSGTEFTLTISSLQPDDF RSEDTAVYYCAKESGRAVADRWGQGTL ATYYCQQYKSYPWTFGQGTKLEIKR VTVSS(SEQ ID NO: 9)(SEQ ID NO: 8)CDR Sequences (Kabat) CDR Sequences (Kabat)CDR-H1 TYAII CDR-L1 RASHSISSWLA(SEQ ID NO: 10) (SEQ ID NO: 13) CDR-H2 AIIPFQDRGQYAQKFQG CDR-L2 KASTLES(SEQ ID NO: 11) (SEQ ID NO: 14) CDR-H3 ESGRAVADR CDR-L3 QQYKSYPWT(SEQ ID NO: 12) (SEQ ID NO: 15) CDR Sequences (IMGT) CDR Sequences (IMGT)CDR-H1 GGTFNTYA CDR-L1 HSISSW(SEQ ID NO: 16) (SEQ ID NO: 19) CDR-H2 IIPFQDRG CDR-L2 KA(SEQ ID NO: 17)CDR-L3 QQYKSYPWTAttorney Docket No. RBT-014WOCDR-H3 AKESGRAVADR (SEQ ID NO: 20)(SEQ ID NO: 18)CDR Sequences (Chothia) CDR Sequences (Chothia)CDR-H1 GGTFNTY CDR-L1 RASHSISSWLA(SEQ ID NO: 21) (SEQ ID NO: 24) CDR-H2 IPFQDR CDR-L2 KASTLES(SEQ ID NO: 22) (SEQ ID NO: 25) CDR-H3 ESGRAVADR CDR-L3 QQYKSYPWT(SEQ ID NO: 23) (SEQ ID NO: 26)

[0077] Table IB: Pfs230 Antibody (RUPA-32) Sequences (VH / VL)Heavy chain variable region Light chain variable region EVQLVESGGGLVKPGGSLRLSCAASGFAF EIVLTQSPATLSLSPGERATLSCRTSQSL STYAMSWVRQAPGKGLEWVSSISESSSYT SISLVWYQQKPGQAPRLLISDASNRAP YYTD S VKGRFTISRDNAKKSLFLHMNSLR GIPARFSASGSGTDFTLTISSLEPEDFAV AEDTAVYYCARSDGLSMIAAPFDYWGQG YYCQQRSNWPPITFGQGTRLEMK ARVTVSS(SEQ ID NO: 28)(SEQ ID NO: 27)CDR Sequences (Kabat) CDR Sequences (Kabat)CDR-H1 TYAMS CDR-L1 RTSQSLSISLV(SEQ ID NO: 29) (SEQ ID NO: 32) CDR-H2 SISESSSYTYYTDSVKG CDR-L2 DASNRAP(SEQ ID NO: 30) (SEQ ID NO: 33) CDR-H3 SDGLSMIAAPFDY CDR-L3 QQRSNWPPIT(SEQ ID NO: 31) (SEQ ID NO: 34) CDR Sequences (IMGT) CDR Sequences (IMGT)CDR-H1 GFAFSTYA CDR-L1 QSLSIS(SEQ ID NO: 35) (SEQ ID NO: 38) CDR-H2 ISESSSYT CDR-L2 DA(SEQ ID NO: 36)CDR-L3 QQRSNWPPITCDR-H3 ARSDGLSMIAAPFDY (SEQ ID NO: 39)(SEQ ID NO: 37)CDR Sequences (Chothia) CDR Sequences (Chothia)CDR-H1 GFAFSTY CDR-L1 RTSQSLSISLV(SEQ ID NO: 40) (SEQ ID NO: 43)CDR-H2 SESSSY CDR-L2 DASNRAPAttorney Docket No. RBT-014WO(SEQ ID NO: 41) (SEQ ID NO: 44) CDR-H3 SDGLSMIAAPFDY CDR-L3 QQRSNWPPIT(SEQ ID NO: 42) (SEQ ID NO: 45)

[0078] In certain embodiments, a Pfs230-binding moiety disclosed herein is an antibody or an antigen-binding fragment thereof that specifically binds to an antigen of Pfs230. In certain embodiments, the antibody or antigen-binding fragment thereof comprises a Fab fragment. In certain embodiments, the Fab fragment is a single-chain Fab (scFab) fragment. In certain embodiments, a scFab that specifically binds to Pfs230 comprises an amino acid sequence of SEQ ID NO: 51 or a variant thereof having at least 80% (e.g., at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identity thereto. In certain embodiments, a scFab that specifically binds to Pfs230 comprises an amino acid sequence of SEQ ID NO: 53 or a variant thereof having at least 80% (e.g., at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identity thereto.

[0079] In certain embodiments, the Pfs230-binding moiety specifically binds to an epitope of a Pfs230 protein, the epitope comprising amino acids E565, T566, T567, E568, S569, G570, D571, A573, V574, S575, E576, D577, Y579, K581, K610, E612, K653, E655, T656, K657, K698, A699, T700, V701, Y703, E724, and Y726 of SEQ ID NO: 50. In certain embodiments, the Pfs230-binding moiety specifically binds to an epitope of a Pfs230 protein, the epitope comprising amino acids D580, K581, Y582, A583, S584, V592, C593, D594, F595, D597, Q598, K600, P601, T602, E603, K607, V608, C611, E612, V632, L635, K716, and N719 of SEQ ID NO: 50. In certain embodiments, the Pfs230-binding moiety specifically binds to an epitope of a Pfs230 protein, the epitope comprising amino acids D580, K581, Y582, A583, S584, N585, V592, C593, D594, F595, D597, Q598, K600, P601, T602, E603, K607, V608, C611, E612, V632, L635, K716, and N719 of SEQ ID NO: 50.

[0080] In certain embodiments, the malaria-associated antigen-binding moiety comprises a scFab comprising heavy and light chain variable regions (e.g., VH and VL or VK) with sequences that have at least 85%, at least 87.5%, 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 to those of the heavy and light chain variable regions of a Pfs230 antibody.

[0081] In certain embodiments, the malaria-associated antigen-binding moiety comprises a scFab comprising heavy chain variable region (e.g., VH) with an amino acid sequence having at least 85%, at least 87.5%, at least 90%, at least 91%, at least 92%, at least 93%, at leastAttorney Docket No. RBT-014WO94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity to SEQ ID NO: 8, and a light chain variable region (e.g., VL or VK) with an amino acid sequence having at least 85%, at least 87.5%, 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 to SEQ ID NO: 9. In certain embodiments, the malaria-associated antigen-binding moiety comprises a scFab comprising an amino acid sequence having at least 85%, at least 87.5%, 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 to SEQ ID NO: 51.

[0082] In certain embodiments, the malaria-associated antigen-binding moiety comprises a scFab comprising heavy chain variable region (e.g., VH) with an amino acid sequence having at least 85%, at least 87.5%, 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 to SEQ ID NO: 21, and a light chain variable region (e.g., VL or VK) with an amino acid sequence having at least 85%, at least 87.5%, 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 to SEQ ID NO: 28. In certain embodiments, the malaria-associated antigen-binding moiety comprises a scFab comprising an amino acid sequence having at least 85%, at least 87.5%, 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 to SEQ ID NO: 53.3, Fc polypeptides

[0083] In certain embodiments, the Fc polypeptide is a single chain Fc (scFc), which comprises two Fc chains linked together by a covalent linker, e.g., via an amino acid linker.

[0084] An IgG Fc chain (e.g., IgGl Fc chain) typically contains two constant heavy domains (CH2 and CH3) and a hinge region connected to the CH2 domain.

[0085] In certain embodiments, the Fc polypeptide comprises one or more IgGl Fc chains (e.g., human IgGl Fc chains or a human Fc chains), that is, the Fc polypeptide comprises an Fc chain that has an amino acid sequence that is substantially similar or identical to that of the chains within a wild type IgGl Fc.

[0086] In certain embodiments, the Fc polypeptide comprises one or more human IgGl Fc chains; that is, the Fc polypeptide comprises an Fc chain that is substantially similar or identical to that of the Fc chains within a wild type human IgGl. In certain embodiments, the wild type human IgGl Fc has an amino acid sequence of SEQ ID NO: 4.Attorney Docket No. RBT-014WO

[0087] In certain embodiments, an Fc polypeptide comprises an Fc chain that comprises the particular residue(s) at certain position(s) specifically described for that Fc chain, but has an amino acid sequence that is otherwise 100% identical to a corresponding Fc chain within a wild type Fc chain, e.g, wild type IgGl Fc chain. In certain embodiments, the Fc polypeptide comprises an Fc chain that has an amino acid sequence that differs by at least one, at least two, at least three, or at least four amino acid residues from the sequence of SEQ ID NO:4. In certain embodiments, the Fc polypeptide comprises an Fc chain that has an amino acid sequence that differs by no more than ten, no more than nine, no more than eight, no more than seven, no more than six, no more than five, or no more than four amino acid residues from the sequence of SEQ ID NO:4.

[0088] In certain embodiments, the Fc polypeptide comprises an Fc chain that has an amino acid sequence having at least 85%, at least 87.5%, 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 to that of SEQ ID NO: 4.4, Linkers

[0089] In certain embodiments, linkers are used within fusion polypeptides and / or within single-chain molecules such as scFcs. In certain embodiments, the linker is an amino acid linker. For example, a linker as employed herein may comprise from about 1 to about 100 amino acid residues, e.g, about 1 to about 70, about 2 to about 70, about 1 to about 30, or about 2 to about 30 amino acid residues. In certain embodiments, the linker comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, or at least 10 amino acid residues.

[0090] In certain embodiments, the linker comprises a glycine-serine sequence, e.g, a (GnS)m sequence (e.g., GGS, GGGS (SEQ ID NO: 6), and / or GGGGS (SEQ ID NO: 7) sequence) that is present in at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, or at least 14 copies within the linker (e.g, GGGGSGGGGSGGGGSGGGGSGGGGSGG (SEQ ID NO: 55)).B. Self-assembled polypeptide complexes

[0091] In one aspect, provided are self-assembled polypeptide complexes (“Multabodies” or “MBs”) comprising a plurality of fusion polypeptides as disclosed herein. Generally, provided self-assembled polypeptide complexes comprise (a) a plurality ofmalaria-associated antigen-binding fusion polypeptides, each malaria-associatedAttorney Docket No. RBT-014WOantigen-binding fusion polypeptide comprising (1) a malaria-associated antigen-binding moiety (e.g., a Pfs230-binding moiety) linked to (2) a nanocage monomer or subunit thereof, and (b) a plurality of Fc fusion polypeptides, each Fc fusion polypeptide comprising (1) an Fc polypeptide (e.g., a wild-type Fc polypeptide) linked to (2) a nanocage monomer or subunit thereof, wherein the Fc polypeptide comprises an Fc chain.

[0092] In certain embodiments, the nanocage monomer is an apoferritin monomer, and each fusion polypeptide within the self-assembled polypeptide complex comprises an apoferritin light chain or a subunit of an apoferritin light chain. In these embodiments, theself-assembled polypeptide complex does not comprise any apoferritin heavy chains, subunits of apoferritin heavy chains, or other apoferritin components capable of binding to iron or capable of ferroxidase activity.

[0093] In certain embodiments, the nanocage monomer or subunit thereof is an apoferritin monomer subunit, and (a) each malaria-associated antigen-binding fusion polypeptide comprises an apoferritin monomer subunit which is N-half apoferritin and each Fc fusion polypeptide comprises an apoferritin monomer subunit which is C-half apoferritin, or (b) each malaria-associated antigen-binding fusion polypeptide comprises an apoferritin monomer subunit which is C-half apoferritin and each Fc fusion polypeptide comprises an apoferritin monomer subunit which is N-half apoferritin.

[0094] In certain embodiments, the self-assembled polypeptide complex comprises between 24 and 48 fusion polypeptides in total. In certain embodiments, the self-assembled polypeptide complex comprises 24 fusion polypeptides in total. In certain embodiments, the self-assembled polypeptide complex comprises more than 24 fusion polypeptides, e.g, at least 26, at least 28, at least 30, at least 32 fusion polypeptides, at least 34 fusion polypeptides, at least 36 fusion polypeptides, at least 38 fusion polypeptides, at least 40 fusion polypeptides, at least 42 fusion polypeptides, at least 44 fusion polypeptides, at least 46 fusion polypeptides, or at least 48 fusion polypeptides in total. In certain embodiments, the self-assembled polypeptide complex comprises about 32 fusion polypeptides.

[0095] In certain embodiments, the self-assembled polypeptide complex comprises at least 4, at least 5, least 6, at least 7, or at least 8 malaria-associated antigen-binding fusion polypeptides.

[0096] In certain embodiments, the self-assembled polypeptide complex comprises at least 4, at least 5, least 6, at least 7, or at least 8 Fc fusion polypeptides.Attorney Docket No. RBT-014WO

[0097] In certain embodiments, the self-assembled polypeptide complex comprises a ratio of approximately 1:1, 11:13, 3:5, 1:2, 7:17, 1:3, 2:7, 5:19, 1:4, 1:5, 1:6, 1:7, 1:8, 1:12, 1:24 of malaria-associated antigen-binding fusion polypeptides to Fc fusion polypeptides.Pharmacokinetic characteristics

[0098] In certain embodiments, when administered to a subject in need thereof, a provided self-assembled polypeptide complex, has one or more pharmacokinetic features similar to that of a reference IgG molecule (e.g., an IgG molecule whose class matches the class of an Fc chain within an Fc polypeptide of an Fc fusion polypeptide within the self-assembled polypeptide complex).C. Methods of treatment and methods of contacting cells

[0099] In one aspect, provided are methods that may be useful for treating, ameliorating, or preventing a disease or a condition, generally comprising a step of administering a self-assembled polypeptide complex of the present disclosure (or a composition thereof) to a subject.

[0100] In certain embodiments, the subject is a mammal, e.g, a human.

[0101] Compositions for administration to subjects generally comprise a self-assembled polypeptide complex as disclosed herein. In certain embodiments, such compositions further comprise a pharmaceutically acceptable excipient.

[0102] Compositions may be formulated for administration for any of a variety of routes of administration, including systemic routes (e.g., oral, inhalation, intranasal, intravenous, intraperitoneal, subcutaneous, or intramuscular administration).

[0103] In certain embodiments, the step of administering results in improvement in one or more clinical outcomes or metrics in the subject.

[0104] For example, in certain embodiments, the subject has an infectious disease. In certain embodiments, the infectious disease is malaria.

[0105] In certain embodiments, the step of administering results in slowing or inhibiting progression of a malaria infection and / or amelioration of one or more symptoms of malaria (e.g., fever, chills, headache, sweating, fatigue, dry cough, muscle pain, back pain, enlarged spleen, nausea, vomiting, abnormal posture, nystagmus, conjugate gaze palsy, opisthotonus, seizures, and coma) by any amount, e.g., 10% or more (e.g., 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or more).Methods of contactingAttorney Docket No. RBT-014WO

[0106] The present disclosure also provides methods of contacting self-assembled polypeptide complexes as disclosed herein with a cell (e.g., a parasitic cell, a mammalian cell infected with a parasitic cell). In certain embodiments, the step of contacting occurs inside a mammalian body, e.g., a human body.

[0107] In certain embodiments, the step of contacting induces apoptosis of the cell. In certain embodiments, a self-assembled polypeptide complex disclosed herein is capable of killing a cell (e.g., parasitic cell or mammalian cell infected with a parasitic cell) with an half-maximal inhibitory concentration (IC50) of no more than 10 nM, no more than 9 nM, no more than 8 nM, no more than 7 nM, no more than 6 nM, no more than 5 nM, no more than 4 nM, or no more than 3 nM. In certain embodiments, the cell expresses Pfs230.EXAMPLESExample 1. Construction and Expression of «-Pfs230 Multabodies (MBs)

[0001] This Example describes the generation of a-Pfs230 Multabodies (MBs) which selfassembled from pluralities of at least two types of apoferritin-containing fusion polypeptides, specifically, MBs which comprise:

[0002] (1) a full-length human apoferritin light chain;

[0003] (2) a malaria-associated antigen-binding fusion polypeptide comprising a subunit of human apoferritin light chain and a single-chain Fab fragment (scFab) that is capable of binding (e.g., specifically binding) to the malaria-associated antigen, Pfs230 (SEQ ID NO: 50);

[0004] (3) an Fc fusion polypeptide comprising a subunit of human apoferritin light chain and either (a) a wild-type (WT) IgGl single chain Fc (scFc) polypeptide or (b) an IgGl scFc polypeptide comprising: an alanine at position 234, an alanine at position 235, an arginine at position 236, an alanine at position 237, and a leucine at position 330 (“LLRAL” mutations), according to EU numbering.

[0005] Genes encoding (1) full-length apoferritin light chain (SEQ ID NO: 1; “hFTL”); (2) a fusion protein which includes (a) a scFab derived from an a-Pfs230, LMIV230-01 (Coehlo et al. Nat. Comm. 12(1): 1750 (2021); SEQ ID NO: 51; see also Table 1A) or (b) a scFab derived from a a-Pfs230, RUPA-32 (Ivanochko et al. Immunity 56(2):420-32 (2023); SEQ ID NO: 53; see also Table IB), fused to the N-terminus of an N-half apoferritin (LMIV230-01 scFab-N hFTL - SEQ ID NO: 52; RUPA-32 scFab-N hFTL - SEQ ID NO: 54) via an amino acid linker (SEQ ID NO: 55), and (3) a scFc polypeptide fused to the N-terminus of a C-half apoferritin (SEQ ID NO: 48 or SEQ ID NO: 49) via an amino acid linker (SEQ ID NO: 55) were prepared, mixed at a molar ratio of 2: 1: 1 and transiently transfected into HEKAttorney Docket No. RBT-014WO293F cells for the production and formation of a Pfs230-targeting MBs. Upon expression, the encoded polypeptides self-assembled to form 8-mer a-Pfs230 MBs.

[0006] MBs were subsequently purified with Protein A columns using Buffer A (20 mM Tris, pH 8, and 150 mM NaCl) and Buffer B (20 mM Tris, pH 8, 3 M MgCl2and 10% glycerol). MBs were concentrated to 500 µL in a 100K concentrator and run on a size exclusion chromatography column (Superose6) with 1X PBS. MBs were adjusted to a concentration of 1 mg / mL, sterile-filtered using a 0.22 µm filter, aliquoted, and stored at -80°C.

[0007] Additionally, parental full-length LMIV230-01 or RUPA-32 monoclonal antibodies (IgGs) were produced with WT IgGl Fc or LLRAL Fc and purified with Protein A columns using Buffer C (IX PBS) and Buffer D (100 mM glycine, pH 2.2) neutralized with 1 M Tris buffer, pH 9. Parental IgGs were concentrated to 500 µL in a 100K concentrator and run on a SEC column (Superose6) with 1X PBS. Antibodies were adjusted to a concentration of 1 mg / mL, sterile-filtered with a 0.22 µm filter, aliquoted, and stored at -80°C.Example 2. Assessment of in vitro parasite inhibitory activity facilitated by «-Pfs230 MBs

[0008] The efficiency of a-Pfs230 MBs in facilitating killing of Plasmodium falciparum (P. falciparum) gametocytes expressing Pfs230 was assessed in a standard membrane feeding assay (SMFA). A transgenic NF54-HGL P. falciparum line expressing GFP: Luc from an hsp70 promoter was employed. a-Pfs230 MBs and full-length LMIV230-01 or RUPA-32 antibodies were generated as described in Example 1. Stage V gametocytes were combined in human type A serum and active complement at 50% hematocrit in the presence of (1) a- Pfs230 MBs with WT scFc, (2) a-Pfs230 MBs with LLRAL scFc, (3) full-length LMIV230- 01 or RUPA-32 antibody with WT Fc, or (4) full-length LMIV230-01 or RUPA-32 antibody with LLRAL Fc, and fed to Anopheles stephensi mosquitos. The MBs were provided at the following concentrations: 0.09117124 nM, 0.28810113 nM, 0.91039958 nM, 2.87686268 nM, 9.09088608 nM, and 28.7272 nM; whereas the full-length antibodies were provided at the following concentrations: 0.66859093 nM, 2.11274733 nM, 6.67628157 nM, 21.0970498 nM, 66.6666772 nM, and 210.6667 nM. Fed mosquitos were subsequently transferred to cages and non-fed mosquitos were removed. Mosquitos were kept for 8 days after feeding and dissected to enumerate oocysts by luciferase assay on the same day. The luminescence signal in infected mosquitos was proportional to the oocyst intensity. Therefore, greater cytotoxicity of the tested molecules was expected to produce a reduced RLU signal.Attorney Docket No. RBT-014WO

[0009] Compared to parental IgGs with WT Fc, a-Pfs230 MBs with WT scFc exhibited a substantially reduced RLU signal at all tested concentrations, especially apparent at highest concentrations, demonstrating the advantageous effect of increased avidity of a-Pfs230 MBs to Pfs230 expressed in gametocytes (FIGs. 1A-1C). Both the parental IgGs with WT Fc and a-Pfs230 MBs with WT scFc produced a dose-dependent reduction in the RLU signal. On the other hand, parental IgGs with LLRAL Fc and a-Pfs230 MBs with LLRAL scFc exhibited minimal reduction in RLU with increasing concentrations of the antibodies or MBs, as compared to their counterparts with WT Fc polypeptides (FIGs. 1A-1C). Moreover, the LLRAL parental IgGs and LLRAL MBs did not substantially differ from one another in their ability to reduce the RLU signal (FIGs. 1A-1C). The above findings were consistent between LMIV230-01-based MBs and IgGs and RUPA-32-based MBs and IgGs (compare FIG. IB against FIG. 1C). Without wishing to be bound by any particular theory, these findings indicate that (1) the anti-malaria effect observed with MBs having WT Fc polypeptides was dependent on intact complement signaling, and (2) the increased anti-parasitic effect observed with WT scFc MBs may relate at least in part to an increased avidity of the MBs to Pfs230 proteins expressed on gametocytes, as compared to WT Fc IgGs. Moreover, the above findings demonstrate that the anti-malaria effect of MBs was consistent regardless of the a-Pfs230 scFab employed, thereby demonstrating the plug-and-play nature of the MB platform, wherein antigen-binding fragments against distinct or overlapping epitopes of a target protein may be substituted without having a negative impact on the anti-malaria effect of the MB.Example 3. Assessment of «-Pfs230 MB and IgG gametocyte binding using a suspension immunofluorescence assay

[0010] To assess whether the LLRAL Fc mutations included in a-Pfs230 MBs and IgGs affect Pfs230 surface antigen binding, a suspension immunofluorescence assay (SIFA) was performed on Stage V gametes from the NF54 line of P. falciparum. Gametogenesis was induced with xanturenic acid and gametocytes were allowed to activate at room temperature for 30 minutes in complete medium (RPMI 1640 containing 25.85 mM HEPES, 382.66 mM hypoxanthine and 2 mM L-glutamine, supplemented with 23.7 mM sodium carbonate and 8.85% pooled human serum blood type A or AB). Freshly isolated gametocytes were counted and concentrated in SIFA buffer (1% heat-inactivated fetal bovine serum, 0.05% sodium azide in PBS). Samples and controls were diluted in SIFA buffer and were subsequently incubated with gametocytes (10,000 gametes per well) in a 96-well V-bottom plate and incubated for 1 h at 4°C. Samples included varying concentrations of (1) a-Pfs230 MBsAttorney Docket No. RBT-014WOcomprising a scFab derived from LMIV230-01 or RUPA-32 with WT scFc, (2) a-Pfs230 MBs comprising a scFab derived from LMIV230-01 or RUPA-32 with LLRAL scFc, (3) full-length LMIV230-01 or RUPA-32 antibody with WT Fc, or (4) full-length LMIV230-01 or RUPA-32 antibody with LLRAL Fc. The MBs were provided at the following concentrations: 0.09117124 nM, 0.28810113 nM, 0.91039958 nM, 2.87686268 nM, 9.09088608 nM, and 28.7272 nM; whereas the full-length antibodies were provided at the following concentrations: 0.66859093 nM, 2.11274733 nM, 6.67628157 nM, 21.0970498 nM, 66.6666772 nM, and 210.6667 nM. Following incubation, gametes were washed 3 times in ice-cold SIFA buffer and stained using Hoechst and Alexa Fluor TM 488 goat anti-human IgG (H+L) secondary antibodies for 1 h at room temperature in the dark. After staining, gametes were washed 3 times in ice-cold SIFA buffer, fixed using 4% paraformaldehyde, and imaged by automated microscopy on a ImageXpress PICO (Molecular Devices). The number and intensity of stained gametes were quantified using Cell Reporter Xpress (CRX) software.

[0011] As shown in FIGs. 2A-2D (LMIV230-01-based constructs) and FIGs.3A-3D (RUPA-32-based constructs), introduction of LLRAL Fc mutations did not lead to systematic reductions in surface antigen binding of MBs and IgGs, as compared to counterpart constructs containing WT Fc polypeptides. The EC50values for antigen-binding between the various tested constructs are provided in Table 4, below.Table 4: EC50malaria antigen-binding values from SIFA experiments Construct EC50(nM) MB (LMIV230-01 scFab) with WT Fc 222IgGl (LMIV230-01) with WT Fc 351MB (LMIV230-01 scFab) with LLRAL Fc 325IgGl (LMIV230-01) with LLRAL Fc 343MB (RUPA-32 scFab) with WT Fc 57IgGl (RUPA-32) with WT Fc 73MB (RUPA-32 scFab) with LLRAL Fc 41IgGl (RUPA-32) with LLRAL Fc 177

[0012] These findings suggest that the functional difference in the anti-malaria effect between MBs and IgGs containing WT Fc polypeptides and those containing LLRAL FcAttorney Docket No. RBT-014WOpolypeptides is associated with the complement-activating effects of the WT Fc polypeptides and not from any differences in Pfs230 antigen binding.SEQUENCE LISTING

[0013] Underlining within fusion sequences indicate linker sequences.

[0014] Bolding within fusion sequences indicate apoferritin or apoferritin subunit sequences.

[0015] Within variable region sequences, underlining and bolding together indicate complementary determining regions sequences.

[0016] Boxed and bolded residues indicate residues that are mutated relative to a reference molecule, e.g. relative to anlgGl Fc.

[0017] SEQ ID NO:! hFTL (human apoferritin light chain)

[0018] MSSQIRQNYSTDVEAAVNSLVNLYLQASYTYLSLGFYFDRDDVALEGVSHFFRELA EEKREGYERLLKMQNQRGGRALFQDIKKPAEDEWGKTPDAMKAAMALEKKLNQALLDLHALG SARTDPHLCDFLETHFLDEEVKLIKKMGDHLTNLHRLGGPEAGLGEYLFERLTLRHD

[0019] SEQ ID NO: 2 N hFTL (N -half human apoferritin light chain)

[0020] MSSQIRQNYSTDVEAAVNSLVNLYLQASYTYLSLGFYFDRDDVALEGVSHFFRELA EEKREGYERLLKMQNQRGGRALFQDIKKPAEDEW

[0021] SEQ ID NO: 3 C hFTL (N-half human apoferritin light chain)

[0022] GKTPDAMKAAMALEKKLNQALLDLHALGSARTDPHLCDFLETHFLDEEVKLIKKMG DHLTNLHRLGGPEAGLGEYLFERLTLRHD

[0023] SEQ ID NO: 4 Wild-type IgGl Fc

[0024] DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0025] SEQ ID NO: 5 Wild-type IgGl scFc

[0026] DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNAttorney Docket No. RBT-014WO WYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS PGS FFLYS KLTVDKSRWQQGNVFS CS VMHEALHNHYTQKS LS LS PGKGGGGSGGGGSGGGGS GGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0027] SEQ ID NO: 6 Example GS linker

[0028] GGGS

[0029] SEQ ID NO: 7 Example GS linker

[0030] GGGGS

[0031] SEQ ID NOs: 8-45 as shown in Tables 1A-1B.

[0032] SEQ ID NO: 46 human IgGl heavy chain constant region sequence

[0033] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELL GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN VFSCSVMHEALHNHYTQKSLSLSPGK

[0034] SEQ ID NO: 47 IgGl Fc LLRAL

[0035] DKTHTCPPCPAPE|AARA|PSVFLFPPKPKDTLMISRTPEVTCWVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALP|L|PIEKTISK AKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS DGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKAttorney Docket No. RBT-014WO

[0036] SEQ ID NO: 48 IgGl scFc LLRAL

[0037] DKTHTCPPCPAPE|AARA|PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALP|L|PIEKTISK AKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS PGS FFLYS KLTVDKSRWQQGNVFS CS VMHEALHNHYTQKS LS LS PGKGGGGSGGGGSGGGGS GGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSDKTHTCP PCPAPE|AARA]PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP|L|PIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD KSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0038] SEQ ID NO: 49 scFc-C_hFTL IgGl LLRAL

[0039] DKTHTCPPCPAPE|AARA|PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALP|L|PIEKTISK AKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS PGS FFLYS KLTVDKSRWQQGNVFS CS VMHEALHNHYTQKS LS LS PGKGGGGSGGGGSGGGGS GGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSDKTHTCP PCPAPE|AARA|PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP|L|PIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD KSRWQQGNVFS CS VMHEALHNHYTQKS LS LS PGKGGGGSGGGGSGGGGSGGGGSGGGGSGGG KTPDAMKAAMALEKKLNQALLDLHALGSARTDPHLCDFLETHFLDEEVKLIKKMGDHLTNLH RLGGPEAGLGEYLFERLTLRHD

[0040] SEQ ID NO: 50 Pfs230 amino acid sequence (UniProtKB ID: P68874.1) MKKIITLKNLFLIILVYIFSEKKDLRCNVIKGNNIKDDEDKRFHLFYYSHNLFKTPETKEKK NKKECFYKNGGIYNLSKEIRMRKDTSVKIKQRTCPFHKEGSSFEMGSKNITCFYPIVGKKER KTLDTI I IKKNVTNDHVVSSDMHSNVQEKNMILIRNIDKENKNDIQNVEEKIQRDTYENKDY ESDDTLIEWFDDNTNEENFLLTFLKRCLMKIFSSPKRKKTWQKKHKSNFFINSSLKYIYMY LTPSDSFNLVRRNRNLDEEDMSPRDNFVIDDEEEEEEEEEEEEEEEEEEEEEEEEEYDDYVY EESGDETEEQLQEEHQEEVGAESSEESFNDEDEDSVEARDGDMIRVDEYYEDQDGDTYDSTI KNEDVDEEVGEEVGEEVGEEVGEEVGEEVGEEVGEEVGEEVGEEEGEEVGEGVGEEVGEEEG EEVGEEEGEYVDEKERQGEIYPFGDEEEKDEGGESFTYEKSEVDKTDLFKFIEGGEGDDVYKAttorney Docket No. RBT-014WO VDGSKVLLDDDTISRVSKKHTARDGEYGEYGEAVEDGENVIKI IRSVLQSGALPSVGVDELD KIDLSYETTESGDTAVSEDSYDKYASNNTNKEYVCDFTDQLKPTESGPKVKKCEVKVNEPLI KVKIICPLKGSVEKLYDNIEYVPKKSPYVVLTKEETKLKEKLLSKLIYGLLISPTVNEKENN FKEGVIEFTLPPVVHKATVFYFICDNSKTEDDNKKGNRGIVEVYVEPYGNKINGCAFLDEDE EEEKYGNQIEEDEHNEKIKMKTFFTQNIYKKNNIYPCYMKLYSGDIGGILFPKNIKSTTCFE EMIPYNKEIKWNKENKSLGNLVNNSWYNKEMNAKYFNVQYVHIPTSYKDTLNLFCSIILKE EESNLISTSYLVYVSINEELNFSLFDFYESFVPIKKTIQVAQKNVNNKEHDYTCDFTDKLDK TVPSTANGKKLFICRKHLKEFDTFTLKCNVNKTQYPNIEIFPKTLKDKKEVLKLDLDIQYQM FSKFFKFNTQNAKYLNLYPYYLIFPFNHIGKKELKNNPTYKNHKDVKYFEQSSVLSPLSSAD SLGKLLNFLDTQETVCLTEKIRYLNLSINELGSDNNTFSVTFQVPPYIDIKEPFYFMFGCNN NKGEGNIGIVELLISKQEEKIKGCNFHESKLDYFNENISSDTHECTLHAYENDI IGENCLET THPNEVEVEVEDAEIYLQPENCFNNVYKGLNSVDITTILKNAQTYNINNKKTPTFLKIPPYN LLEDVEISCQCTIKQVVKKIKVI ITKNDTVLLKREVQSESTLDDKIYKCEHENFINPRVNKT FDENVEYTCNIKIENFFNYIQIFCPAKDLGIYKNIQMYYDIVKPTRVPQFKKFNNEELHKLI PNSEMLHKTKEMLILYNEEKVDLLHFYVFLPIYIKDIYEFNIVCDNSKTMWKNQLGGKVIYH ITVSKREQKVKGCSFDNEHAHMFSYNKTNVKNCI IDAKPKDLIGFVCPSGTLKLTNCFKDAI VHTNLTNINGI LYLKNNLANFTYKHQFNYME I PALMDND I S FKC I CVDLKKKKYNVKS PLGP KVLRALYKKLNIKFDNYVTGTDQNKYLMTYMDLHLSHKRNYLKELFHDLGKKKPADTDANPE SIIESLSINESNESGPFPTGDVDAEHLILEGYDTWESLYDEQLEEVIYNDIESLELKDIEQY VLQVNLKAPKLMMSAQIHNNRHVCDFSKNNLIVPESLKKKEELGGNPVNIHCYALLKPLDTL YVKCPTSKDNYEAAKVNISENDNEYELQVISLIEKRFHNFETLESKKPGNGDVVVHNGVVDT GPVLDNSTFEKYFKNIKIKPDKFFEKVINEYDDTEEEKDLESILPGAIVSPMKVLKKKDPFT SYAAFVVPPIVPKDLHFKVECNNTEYKDENQYISGYNGI IHIDISNSNRKINGCDFSTNNSS ILTSSVKLVNGETKNCEININNNEVFGIICDNETNLDPEKCFHEIYSKDNKTVKKFREVIPN IDIFSLHNSNKKKVAYAKVPLDYINKLLFSCSCKTSHTNTIGTMKVTLNKDEKEEEDFKTAQ GIKHNNVHLCNFFDNPELTFDNNKIVLCKIDAELFSEVIIQLPIFGTKNVEEGVQNEEYKKF SLKPSLVFDDNNNDIKVIGKEKNEVSISLALKGVYGNRIFTFDKNGKKGEGISFFIPPIKQD TDLKFIINETIDNSNIKQRGLIYIFVRKNVSENSFKLCDFTTGSTSLMELNSQVKEKKCTVK IKKGDIFGLKCPKGFAIFPQACFSNVLLEYYKSDYEDSEHINYYIHKDKKYNLKPKDVIELM DENFRELQNIQQYTGISNITDVLHFKNFNLGNLPLNFKNHYSTAYAKVPDTFNSIINFSCNC YNPEKHVYGTMQVESDNRNFDNIKKNENVIKNFLLPNIEKYALLLDDEERQKKIKQQQEEEQ QEQILKDQDDRLSRHDDYNKNHTYILYDSNEHICDYEKNESLISTLPNDTKKIQKSICKINA KALDVVTIKCPHTKNFTPKDYFPNSSLITNDKKIVITFDKKNFVTYIDPTKKTFSLKDIYIQ SFYGVSLDHLNQIKKIHEEWDDVHLFYPPHNVLHNWLNNHIVNLSSALEGVLFMKSKVTGDAttorney Docket No. RBT-014WO ETATKKNTTLPTDGVSSILIPPYVKEDITFHLFCGKSTTKKPNKKNTSLALIHIHISSNRNI IHGCDFLYLENQTNDAISNNNNNSYSIFTHNKNTENNLICDISLIPKTVIGIKCPNKKLNPQ TCFDEVYYVKQEDVPSKTITADKYNTFSKDKIGNILKNAISINNPDEKDNTYTYLILPEKFE EELIDTKKVLACTCDNKYI IHMKIEKSTMDKIKIDEKKTIGKDICKYDVTTKVATCEI IDTI DSSVLKEHHTVHYSITLSRWDKLIIKYPTNEKTHFENFFVNPFNLKDKVLYNYNKPINIEHI LPGAITTDIYDTRTKIKQYILRIPPYVHKDIHFSLEFNNSLSLTKQNQNIIYGNVAKIFIHI NQGYKEIHGCDFTGKYSHLFTYSKKPLPNDDDICNVTIGNNTFSGFACLSHFELKPNNCFSS VYDYNEANKVKKLFDLSTKVELDHIKQNTSGYTLSYIIFNKESTKLKFSCTCSSNYSNYTIR ITFDPNYI IPEPQSRAI IKYVDLQDKNFAKYLRKL

[0041] SEQ ID NO: 51 Amino acid sequence of scFab derived from LMIV230-01 antibody

[0042] MEFGLSWVFLVALFRGVQSDIVMTQSPASLALSLGERATLYCRASHSISSWLAWYQ QKPGKAPKLLIYKASTLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYKSYPWTFG QGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGG SGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSQ VQLVQSGAEVKKPGSS VKVSCKVSGGTFNTYAI I WVRQAPGQGLEWMGAI I PFQDRGQYAQK FQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAKESGRAVADRWGQGTLVTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC

[0043] SEQ ID NO: 52 LMIV230-01 scFab-N hFTL

[0044] MEFGLSWVFLVALFRGVQSDIVMTQSPASLALSLGERATLYCRASHSISSWLAWYQ QKPGKAPKLLIYKASTLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYKSYPWTFG QGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGG SGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSQ VQLVQSGAEVKKPGSS VKVSCKVSGGTFNTYAI I WVRQAPGQGLEWMGAI I PFQDRGQYAQK FQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAKESGRAVADRWGQGTLVTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSGGGGSGGGGSGGGGSGGGGSGGMAttorney Docket No. RBT-014WO SSQIRQNYSTDVEAAVNSLVNLYLQASYTYLSLGFYFDRDDVALEGVSHFFRELAEEKREGY ERLLKMQNQRGGRALFQD I KKP AEDEW

[0045] SEQ ID NO: 53 Amino acid sequence of scFab derived from RUPA-32 antibody EIVLTQSPATLSLSPGERATLSCRTSQSLSISLVWYQQKPGQAPRLLISDASNRAPGIPARF SASGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPITFGQGTRLEMKRTVAAPSVFIFPPSDE QLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGG GS GGGGS GGGGS GGGGS GGGGS GGGGS GGGGS GGGGS E VQLVE S GGGL VKPGGS LRLS CAAS GFAFSTYAMSWVRQAPGKGLEWVSSISESSSYTYYTDSVKGRFTISRDNAKKSLFLHMNSLR AEDTAVYYCARSDGLSMIAAPFDYWGQGARVTVSSASTKGPSVFPLAPSSKSTSGGTAALGC LVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYICNVNHKP SNTKVDKKVEPKSC

[0046] SEQ ID NO: 54 RUPA-32 scFab-N hFTL

[0047] EIVLTQSPATLSLSPGERATLSCRTSQSLSISLVWYQQKPGQAPRLLISDASNRAP GIPARFSASGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPITFGQGTRLEMKRTVAAPSVFI FPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLS KADYEKHKVYACE VTHQGLS S PVTKS FNRGECGGGGSGGGGSGGGGSGGGGSGGGGSGG GGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSEVQLVE S GGGL VKPGGS LR LSCAASGFAFSTYAMSWVRQAPGKGLEWVSSISESSSYTYYTDSVKGRFTISRDNAKKSLFL HMNSLRAEDTAVYYCARSDGLSMIAAPFDYWGQGARVTVSSASTKGPSVFPLAPSSKSTSGG TAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCGGGGSGGGGSGGGGSGGGGSGGGGSGGMSSQIRQNYSTDVEA AVNSLVNLYLQASYTYLSLGFYFDRDDVALEGVSHFFRELAEEKREGYERLLKMQNQRGGRA LFQDIKKPAEDEW

[0048] SEQ ID NO: 55 Example GS linker

[0049] GGGGSGGGGSGGGGSGGGGSGGGGSGG

[0050] SEQ ID NO: 56 scFc-C_hFTL wild-type IgGlAttorney Docket No. RBT-014WO

[0051] DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDS PGS FFLYS KLTVDKSRWQQGNVFS CS VMHEALHNHYTQKS LS LS PGKGGGGSGGGGSGGGGS GGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSGGGGSDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKT KPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVD KSRWQQGNVFS CS VMHEALHNHYTQKS LS LS PGKGGGGSGGGGSGGGGSGGGGSGGGGSGGG KTPDAMKAAMALEKKLNQALLDLHALGSARTDPHLCDFLETHFLDEEVKLIKKMGDHLTNLH RLGGPEAGLGEYLFERLTLRHDEQUIVALENTS / OTHER EMBODIMENTS

[0108] While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure that come within known or customary practice within the art to which the invention pertains and may be applied to the essential features herein before set forth.

Claims

Attorney Docket No. RBT-014WOCLAIMS1. A fusion polypeptide comprising: (1) a malaria-associated antigen-binding moiety and (2) a nanocage monomer or subunit thereof,wherein the malaria-associated antigen-binding moiety comprises an antibody or antigen-binding fragment thereof comprising a heavy chain variable region (VH) comprising a CDR-H1, CDR-H2, and CDR-H3 having sequences which each differ by at most two amino acid residues from the CDR-H1, CDR-H2, and CDR-H3 sequences of a Pfs230 antibody, respectively; and / or a light chain variable region (VL or VK) comprising a CDR-L1, CDR-L2, and CDR-L3 having sequences which each differ by at most two amino acid residues from the CDR-L1, CDR-L2, and CDR-L3 sequences of a Pfs230 antibody, respectively.

2. A fusion polypeptide comprising: (1) a malaria-associated antigen-binding moiety and (2) a nanocage monomer or subunit thereof, wherein the malaria-associated antigen-binding moiety is capable of binding to an epitope of Pfs230.

3. The fusion polypeptide of claim 2, wherein the epitope comprises(a) amino acid residues E565, T566, T567, E568, S569, G570, D571, A573, V574, S575, E576, D577, Y579, K581, K610, E612, K653, E655, T656, K657, K698, A699, T700, V701, Y703, E724, and Y726 of SEQ ID NO: 50; or(b) amino acid residues D580, K581, Y582, A583, S584, V592, C593, D594, F595, D597, Q598, K600, P601, T602, E603, K607, V608, C611, E612, V632, L635, K716, and N719 of SEQ ID NO: 50.

4. The fusion polypeptide of claim 3, wherein the epitope of (b) further comprises an N585 amino acid residue.

5. The fusion polypeptide of any one of claims 1-4, wherein the malaria-associated antigen-binding moiety comprises an antibody or antigen-binding fragment thereof.

6. The fusion polypeptide of claim 5, wherein the antibody or antigen-binding fragment thereof comprises an immunoglobulin heavy chain variable domain (VH) and an immunoglobulin light chain variable domain (VL or VK).Attomey Docket No. RBT-014WO7. The fusion polypeptide of claim 6, wherein the VH comprises complementarity -determining regions (CDRs) CDR-H1, CDR-H2, and CDR-H3 having sequences which each differ by at most two amino acid residues from the sequences of SEQ ID NOs: 10, 11, and 12, respectively; and the VL or VK comprises CDRs CDR-L1, CDR-L2, and CDR-L3 having sequences which each differ by at most two amino acid residues from the sequences of SEQ ID NOs: 13, 14, and 15, respectively.

8. The fusion polypeptide of claim 7, wherein the CDR-H1, CDR-H2, and CDR-H3 have sequences identical to those of SEQ ID NOs: 10, 11, and 12, respectively; and the CDR-Ll, CDR-L2, and CDR-L3 have sequences identical to those of SEQ ID NOs: 13, 14, andl5, respectively, except for one or two amino acid substitutions total across all six CDRs.

9. The fusion polypeptide of claim 8, wherein the CDR-H1, CDR-H2, and CDR-H3 have sequences identical to those of SEQ ID NOs: 10, 11, and 12, respectively; and the CDR-Ll, CDR-L2, and CDR-L3 have sequences identical to those of SEQ ID NOs: 13, 14, 15, respectively.

10. The fusion polypeptide of any one of claims 6-9, wherein the VH comprises a sequence having at least 85% identity to the sequence of SEQ ID NO: 8; and the VL or VK comprises a sequence having at least 85% identity to the sequence of SEQ ID NO: 9.

11. The fusion polypeptide of claim 6, wherein the VH comprises CDRs CDR-H1, CDR-H2, and CDR-H3 having sequences which each differ by at most two amino acid residues from the sequences of SEQ ID NOs: 29, 30, and 31, respectively; and the VL or VK comprises CDRs CDR-L1, CDR-L2, and CDR-L3 having sequences which each differ by at most two amino acid residues from the sequences of SEQ ID NOs: 32, 33, and 34, respectively.

12. The fusion polypeptide of claim 11, wherein the CDR-H1, CDR-H2, and CDR-H3 have sequences identical to those of SEQ ID NOs: 29, 30, and 31, respectively; andthe CDR-Ll, CDR-L2, and CDR-L3 have sequences identical to those of SEQ ID NOs: 32, 33, and 34, respectively, except for one or two amino acid substitutions total across all six CDRs.

13. The fusion polypeptide of claim 12, wherein the CDR-H1, CDR-H2, and CDR-H3 have sequences identical to those of SEQ ID NOs: 29, 30, and 31, respectively; and the CDR-Attorney Docket No. RBT-014WOLI, CDR-L2, and CDR-L3have sequences identical to those of SEQ ID NOs: 32, 33, and 34, respectively.

14. The fusion polypeptide of any one of claims 6 or 11-13, wherein the VH comprises a sequence having at least 85% identity to the sequence of SEQ ID NO: 27; and the VL or VK comprises a sequence having at least 85% identity to the sequence of SEQ ID NO: 28.

15. The fusion polypeptide of any one of claims 5-14, wherein the antibody or antigenbinding fragment thereof comprises or consists of a Fab fragment.

16. The fusion polypeptide of claim 15, wherein the Fab fragment is a single-chain Fab fragment (scFab).

17. The fusion polypeptide of claim 5, wherein the antibody or antigen-binding fragment thereof comprises or consists of an scFab which comprises a sequence having at least 80% identity to the sequence of SEQ ID NO: 51.

18. The fusion polypeptide of claim 5, wherein the antibody or antigen-binding fragment thereof comprises or consists of an scFab which comprises a sequence having at least 80% identity to the sequence of SEQ ID NO: 53.

19. The fusion protein of claim 1 or 2, wherein the Pfs230 antibody is LMIV230-01.

20. The fusion protein of claim 1 or 2, wherein the Pfs230 antibody is RUPA-32.

21. The fusion polypeptide of any one of claims 1-20, wherein the nanocage monomer or subunit thereof is an apoferritin monomer or subunit thereof.

22. The fusion polypeptide of claim 21, wherein the apoferritin monomer is a human apoferritin monomer r.

23. The fusion polypeptide of claim 21 or 22, wherein the apoferritin monomer is an apoferritin light chain.Attorney Docket No. RBT-014WO24. The fusion polypeptide of any one of claims 1-23, wherein the malaria-associated antigen-binding moiety is linked via an amino acid linker to the nanocage monomer or subunit thereof.

25. The fusion polypeptide of any one of claims 1-23, wherein the malaria-associated antigen-binding moiety is linked via the N-terminus of the nanocage monomer or subunit thereof.

26. A self-assembled polypeptide complex comprising:(a) a plurality of malaria-associated antigen-binding fusion polypeptides, each fusion polypeptide being a fusion polypeptide of any one of claims 1-25; and (b) a plurality of Fc fusion polypeptides, each Fc fusion polypeptide comprising (1) an Fc polypeptide and (2) a nanocage monomer or subunit thereof.

27. The self-assembled polypeptide complex of claim 26, wherein, within each Fc fusion polypeptide, the Fc polypeptide is linked via an amino acid linker to the nanocage monomer or subunit thereof.

28. The self-assembled polypeptide complex of claim 27, wherein the Fc polypeptide is linked via the N-terminus of the nanocage monomer or subunit thereof.

29. A self-assembled polypeptide complex comprising:(a) a plurality of malaria-associated antigen-binding fusion polypeptides, each fusion polypeptide being a fusion polypeptide of any one of claims 1-25; and (b) a plurality of Fc fusion polypeptides, each Fc fusion polypeptide comprising (1) an Fc polypeptide linked via an amino acid linker to the N-terminus of (2) a nanocage monomer subunit.

30. The self-assembling polypeptide complex of claim 29, wherein the nanocage monomer subunit is a C-half nanocage monomer.

31. The self-assembling polypeptide complex of claim 30, wherein the C-half nanocage monomer is a C-half apoferritin and the nanocage monomers within the malaria-associated antigen-binding fusion polypeptide are each an apoferritin monomer or a subunit thereof.Attorney Docket No. RBT-014WO32. The self-assembled polypeptide complex of claim 31, wherein the apoferritin monomer is a human apoferritin monomer.

33. The self-assembled polypeptide complex of claim 31 or 32, wherein the apoferritin monomer is an apoferritin light chain.

34. The self-assembled polypeptide complex of claim 33, which does not comprise any apoferritin heavy chains or subunits of apoferritin heavy chains.

35. The self-assembled polypeptide complex of any one of claims 30-34, which does not comprise or enclose iron.

36. The self-assembled polypeptide complex of claim 29, comprising a 2: 1: 1 ratio of a full-length human apoferritin; an Fc polypeptide fused to a first apoferritin monomer subunit; and a malaria-associated antigen-binding moiety fused to a second apoferritin monomer subunit.

37. The self-assembled polypeptide complex of claim 29, comprising a 2: 1: 1 ratio of a full-length human apoferritin; an Fc polypeptide fused to C-half apoferritin; and a malaria-associated antigen-binding moiety fused to N-half apoferritin.

38. The self-assembled polypeptide complex of claim 29, comprising a 2: 1: 1 ratio of a full-length human apoferritin; an Fc polypeptide fused to the N-terminus of C-half apoferritin; and a malaria-associated antigen-binding moiety fused to the N-terminus of N-half apoferritin.

39. The self-assembled polypeptide complex of any one claims 26-38, wherein the Fc polypeptide is an IgGl Fc polypeptide.

40. A pharmaceutical composition comprising a self-assembled polypeptide complex of any one of claims 26-39 and a pharmaceutically acceptable excipient.Attorney Docket No. RBT-014WO41. A method of treating, ameliorating, or preventing a disease or condition, the method comprising administering to a subject in need thereof the self-assembled polypeptide complex of any of claims 26-39 or the pharmaceutical composition of claim 40.

42. The method of claim 41, wherein the subject is a mammal.

43. The method of claim 42, wherein the subject is human.

44. The method of any one of claims 41-43, wherein the disease or condition is malaria.

45. The method of claim 44, wherein the malaria is caused by an infection with P. falciparum.

46. Use of the self-assembled polypeptide complex of any one of claims 26-39 or the pharmaceutical composition of claim 40 to treat, ameliorate, or prevent a disease or condition in a subject in need thereof.

47. The use of claim 46, wherein the disease or condition is malaria.

48. The use of claim 47, wherein the malaria is caused by an infection with P. falciparum.

49. A method comprising a step of contacting a cell with a self-assembled polypeptide complex of any one of claims 26-39.

50. The method of claim 49, wherein the cell is infected with a parasite.

51. The method of claim 50, wherein the parasite is P. falciparum.

52. The method of claim 51, wherein the step of contacting results in killing of the infected cell.

53. The method of any one of claims 49-52, wherein the infected cell expresses Pfs230.Attorney Docket No. RBT-014WO54. The method of any one of claims 49-53, wherein the self-assembled polypeptide complex is capable of killing a parasite with an half-maximal inhibitory concentration (IC50) of no more than 10 nM, no more than 9 nM, no more than 8 nM, no more than 7 nM, no more than 6 nM, no more than 5 nM, no more than 4 nM, or no more than 3 nM.

55. The method of any one of claims 49-54, wherein the step of contacting occurs inside a mammal having an infection.

56. The method of claim 54, wherein the mammal is human.

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