Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
  • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
  • C07K16/20—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans from protozoa
  • C07K16/205—Plasmodium
• • 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
• • 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
  • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
  • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
  • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
  • C07K2317/33—Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
  • C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
  • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
  • C07K2317/732—Antibody-dependent cellular cytotoxicity [ADCC]
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
  • C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K2317/00—Immunoglobulins specific features
  • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
  • C07K2317/94—Stability, e.g. half-life, pH, temperature or enzyme-resistance
• • 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 antibody is a recombinant anti-circumsporozoite (CSP) antibody.
• the recombinant antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), wherein the light chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 3; and the heavy chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 4, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 5, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 6
• the VL comprises the amino acid sequence set forth in SEQ ID NO: 163, and the VH comprises the amino acid sequence set forth in SEQ ID NO: 164; the VL comprises the amino acid sequence set forth in SEQ ID NO: 33, and the VH comprises the amino acid sequence set forth in SEQ ID NO: 34; the VL comprises the amino acid sequence set forth in SEQ ID NO: 43, and the VH comprises the amino acid sequence set forth in SEQ ID NO: 44; the VL comprises the amino acid sequence set forth in SEQ ID NO: 53, and the VH comprises the amino acid sequence set forth in SEQ ID NO: 54; the VL comprises the amino acid sequence set forth in SEQ ID NO: 63, and the VH comprises the amino acid sequence set forth in SEQ ID NO: 64; the VL comprises the amino acid sequence set forth in SEQ ID NO: 73, and the VH comprises the amino acid sequence set forth in SEQ ID NO: 74; the VL comprises the amino acid sequence set forth in SEQ ID NO: 83, and the VH comprises
• the recombinant antibody comprises a light chain (LC) and a heavy chain (HC).
• the LC comprises the amino acid sequence set forth in SEQ ID NO: 27; the amino acid sequence set forth in SEQ ID NO: 37; the amino acid sequence set forth in SEQ ID NO: 47; the amino acid sequence set forth in SEQ ID NO: 57; the amino acid sequence set forth in SEQ ID NO: 67; the amino acid sequence set forth in SEQ ID NO: 77; the amino acid sequence set forth in SEQ ID NO: 87; the amino acid sequence set forth in SEQ ID NO: 97; the amino acid sequence set forth in SEQ ID NO: 107; the amino acid sequence set forth in SEQ ID NO: 117; the amino acid sequence set forth in SEQ ID NO: 127; the amino acid sequence set forth in SEQ ID NO: 137; the amino acid sequence set forth in SEQ ID NO: 147; the amino acid sequence set forth in SEQ ID NO: 157; the amino acid sequence set forth in SEQ ID NO: 27; the amino acid sequence
• the present disclosure provides a recombinant anti circumsporozoite (CSP) antibody comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein the light chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 205, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 206, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 207; and the heavy chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 208, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 209, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 210.
• CSP recombinant anti circumsporozoite
• the recombinant antibody binds to a NANP repeat region. In certain embodiments, the recombinant antibody binds to a polypeptide comprising or consisting of the amino acid sequence set forth in SEQ ID NO: 280. In certain embodiments, the recombinant antibody comprises at least one modification relative to the native AB-000224 variable heavy chain amino acid sequence set forth in SEQ ID NO: 14. In certain embodiments, the recombinant antibody comprises at least one modification relative to the native AB-000224 variable light chain amino acid sequence set forth in SEQ ID NO: 13.
• the present disclosure provides a method of preventing and/or treating malaria in a subject in need thereof, comprising administering an effective amount of the antibody disclosed herein or of the composition disclosed herein.
• the subject is a pediatric patient.
• Figure 6 shows the ASN numbering system for the light chain of AB-000224.
• Figure 7 shows the ASN numbering system for the heavy chain of AB-000224.
• Figures 11 A-l 1C illustrate the survival rate of mice following administration of anti-CSP antibody variants disclosed herein and exposure to mosquitoes infected with chimeric P. berghei expressing P. falciparum CSP protein.
• Figures 11D-F show concentration of human antibodies that were circulating in the mice at the time of infection as determined by ELISA for the assays shown in Figures 11A-C, respectively.
• AB-001245 is a non-malaria-specific antibody that was used as a negative control.
• AB-000317 is a positive control.
• the term “about” or “approximately” refers to the usual error range for the respective value readily known to the skilled person in this technical field, for example, ⁇ 20%, ⁇ 10%, or ⁇ 5%, are within the intended meaning of the recited value.
• anti-CSP antibody and “CSP antibody” are used synonymously and refer to an antibody that binds to Plasmodium falciparum circumsporozoite (CSP) antigen.
• V-region refers to an antibody variable region domain comprising the segments of Framework 1, CDR1, Framework 2, CDR2, Framework 3, CDR3, and Framework 4.
• the heavy chain V-region, VH is a consequence of rearrangement of a V-gene (HV), a D-gene (HD), and a J-gene (HJ), in what is known as V(D)J recombination during B-cell differentiation.
• the light chain V-region, VL is a consequence of the rearrangement of a V-gene (LV) and a J- gene.
• VH and “heavy chain variable” refer to the heavy chain V-region of an antibody.
• the terms “VL” and “light chain variable” refer to the light chain V-region of an antibody.
• ASN Antibody Structural Numbering
• ASN is a numbering system developed based on the AHo numbering system (Honegger & Pluckthun, J. Mol. Biol. 309:657-670 (2001)) defined by Annemarie Honegger for the variable region, but extended to include constant domains.
• Figures 6-7 illustrates ASN numbering for AB-000224-LS light and heavy chain, respectively.
• Figures 8- 9 illustrates ASN numbering for AB-007088-LS light and heavy chain, respectively.
• one or more amino acids can be deleted from the N-terminus or C-terminus of the Fc region of an immunoglobulin without substantial loss of biological function.
• Such variants can be selected according to general rules known in the art to have minimal effect on activity (see, e.g., Bowie, et al., Science 247:306-1310, 1990).
• a single amino acid substitution S228P according to Kabat numbering; designated IgG4Pro
• IgG4Pro a single amino acid substitution
• IgG4Pro an amino acid substitution
• the Fc region includes substitutions that improve pharmacokinetics properties of an antibody, e.g., increased serum half-life.
• substitutions of the Fc region can be found in U.S. Patent No. 8,088,376, the content of which is incorporated by reference in its entirety.
• an "improved" KD refers to a lower KD.
• an antibody of the present disclosure has a KD of less than 5 x 10 5 M, less than 10 5 M, less than 5 x 10 6 M, less than 10 6 M, less than 5 x 10 7 M, less than 10 7 M, less than 5 x 10 8 M, less than 10 8 M, less than 5 x 10 9 M, less than 10 9 M, less than 5 xlO 10 M, less than 10 10 M, less than 5 x 10 U M, less than 10 U M, less than 5 x 10 12 M, less than 10 12 M, less than 5 x 10 13 M, less than 10 13 M, less than 5 x 10 14 M, less than 10 14 M, less than 5 x 10 15 M, or less than 10 15 M or lower as measured as a monovalent antibody, such as a monovalent Fab.
• monovalent molecule refers to a molecule that has one antigen binding site, e.g, a Fab or scFv.
• identity in the context of two or more polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues that are the same (e.g., at least 70%, at least 75%, at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher) identity over a specified region, e.g., the length of the two sequences, when compared and aligned for maximum correspondence over a comparison window or designated region.
• the dipeptide DG poses a medium risk of development liability.
• D may also exhibit low risk of development liability for other successor residues, e.g., N, H, or P.
• an Fc region of the heavy chain of a variant of an anti-CSP antibody AB- 000224 disclosed herein can include an isoleucine at position 250, a tyrosine at position 252, an isoleucine at position 259, a glutamine at position 307, a phenylalanine at position 308, a leucine at position 319, a leucine at position 428, a histidine at position 434, a phenylalanine at position 434, an alanine at position 434, a serine at position 434, a methionine at position 434, or a combination thereof, wherein the numbering is defined by EU index as in Kabat.
• the VL of the variant of an anti-CSP antibody includes at least one, at least two, or at least three mutations, e.g., substitutions, in the framework of the amino acid sequence set forth in SEQ ID NO: 13.
• An exemplary nucleic acid sequence of SEQ ID NO: 13 is set forth in SEQ ID NO: 15.
• the VL of the variant of an anti-CSP antibody includes a substitution at position 1 of the SEQ ID NO: 13.
• the substitution is E1Q.
• the VL of the variant of an anti-CSP antibody includes a substitution at position 4 of the SEQ ID NO: 13.
• the substitution is R44T.
• a variant of an anti-CSP antibody includes one, two, or three CDRs of a VEl sequence of Table 5.
• a variant of an anti-CSP antibody includes at least one mutation, e.g., substitutions, and no more than 10, 20, 30, 40, or 50 mutations in the VH amino acid sequence set forth in SEQ ID NO: 14.
• the VH of the variant of an anti-CSP antibody includes at least one, at least two, or at least three mutations, e.g., substitutions, in the CDR1 amino acid sequence set forth in SEQ ID NO: 4.
• the VH of the variant of an anti-CSP antibody includes a substitution at position 80 of the SEQ ID NO: 14. In certain embodiments, the substitution is I80T. In certain embodiments, the VH of the variant of an anti-CSP antibody includes a substitution at position 90 of the SEQ ID NO: 14. In certain embodiments, the substitution is T90A. In certain embodiments, the VH of the variant of an anti-CSP antibody includes a substitution at position 99 of the SEQ ID NO: 14. In certain embodiments, the substitution is T99A.
• the heavy chain of the variant of an anti-CSP antibody includes a substitution at position 444 of the SEQ ID NO: 18. In certain embodiments, the substitution is N444S. In certain embodiments, the heavy chain of the variant of an anti-CSP antibody includes a substitution at position 438 of the SEQ ID NO: 18 and a substitution at position 444 of the SEQ ID NO: 18. In certain embodiments, the substitutions are M438L and N444S. In certain embodiments, the heavy chain of the variant of an anti-CSP antibody has an amino acid sequence set forth in SEQ ID NO: 19. An exemplary nucleic acid sequence of SEQ ID NO: 19 is set forth in SEQ ID NO: 22.
• the anti-CSP antibody variant comprises a light chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 23, as shown in Table 6.
• the anti-CSP antibody variant comprises a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 24, as shown in Table 6.
• the anti-CSP antibody variant comprises a light chain having the amino acid sequence set forth in SEQ ID NO: 27, as shown in Table 6.
• the anti-CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 28, as shown in Table 6.
• the anti-CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 29, as shown in Table 6.
• Exemplary nucleic acid sequences of SEQ ID NOS: 23, 24, 27, 28, and 29 are provided in Table 6 below.
• the anti- CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 68, as shown in Table 10. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 69, as shown in Table 10. Exemplary nucleic acid sequences of SEQ ID NOS: 63, 64, 67, 68, and 69 are provided in Table 10 below.
• the anti-CSP antibody variant comprises a light chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 73, as shown in Table 11.
• the anti-CSP antibody variant comprises a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 74, as shown in Table 11.
• the anti-CSP antibody variant comprises a light chain having the amino acid sequence set forth in SEQ ID NO: 77, as shown in Table 11.
• the anti- CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 78, as shown in Table 11. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 79, as shown in Table 11. Exemplary nucleic acid sequences of SEQ ID NOS: 73, 74, 77, 78, and 79 are provided in Table 11 below. Table 11
• Antibody ID AB-000224.006
• the anti-CSP antibody variant comprises a light chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 83, as shown in Table 12.
• the anti-CSP antibody variant comprises a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 84, as shown in Table 12.
• the anti-CSP antibody variant comprises a light chain having the amino acid sequence set forth in SEQ ID NO: 87, as shown in Table 12.
• the anti- CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 88, as shown in Table 12. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 89, as shown in Table 12. Exemplary nucleic acid sequences of SEQ ID NOS: 83, 84, 87, 88, and 89 are provided in Table 12 below. Table 12
• the anti-CSP antibody variant comprises a light chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 93, as shown in Table 13.
• the anti-CSP antibody variant comprises a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 94, as shown in Table 13.
• the anti-CSP antibody variant comprises a light chain having the amino acid sequence set forth in SEQ ID NO: 97, as shown in Table 13.
• the anti- CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 98, as shown in Table 13. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 99, as shown in Table 13. Exemplary nucleic acid sequences of SEQ ID NOS: 93, 94, 97, 98, and 99 are provided in Table 13 below.
• the anti-CSP antibody variant comprises a light chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 103, as shown in Table 14.
• the anti-CSP antibody variant comprises a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 104, as shown in Table 14.
• the anti-CSP antibody variant comprises a light chain having the amino acid sequence set forth in SEQ ID NO: 107, as shown in Table 14.
• the anti- CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 108, as shown in Table 14. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 109, as shown in Table 14. Exemplary nucleic acid sequences of SEQ ID NOS: 103, 104, 107, 108, and 109 are provided in Table 14 below.
• the anti- CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 118, as shown in Table 15. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 119, as shown in Table 15. Exemplary nucleic acid sequences of SEQ ID NOS: 113, 114, 117, 118, and 119 are provided in Table 15 below. Table 15
• the anti- CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 148, as shown in Table 18. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 149, as shown in Table 18. Exemplary nucleic acid sequences of SEQ ID NOS: 143, 144, 147, 148, and 149 are provided in Table 18 below.
• the anti-CSP antibody variant comprises a light chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 163, as shown in Table 20.
• the anti-CSP antibody variant comprises a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 164, as shown in Table 20.
• the anti-CSP antibody variant comprises a light chain having the amino acid sequence set forth in SEQ ID NO: 167, as shown in Table 20.
• the anti- CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 178, as shown in Table 21. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 179, as shown in Table 21. Exemplary nucleic acid sequences of SEQ ID NOS: 173, 174, 177, 178, and 179 are provided in Table 21 below.
• the anti-CSP antibody variant comprises a light chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 195, as shown in Table 22.
• the anti-CSP antibody variant comprises a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 196, as shown in Table 22.
• the anti-CSP antibody variant comprises a light chain having the amino acid sequence set forth in SEQ ID NO: 199, as shown in Table 22.
• a variant of an anti-CSP antibody AB-007088 disclosed herein has at least one modification, e.g., substitution, relative to the native AB-007088 variable heavy chain amino acid sequence (SEQ ID NO: 196) or variable light chain amino acid sequence (SEQ ID NO: 195), and has improved developability, e.g., decreased heterogeneity, increased yield, increased stability, improved net charges to improve pharmacokinetics, and/or reduced immunogenicity.
• a VH region or a VL region of such a variant of an anti-CSP antibody AB-007088 disclosed herein has at least two, three, four, five, or six, or more modifications, e.g, substitutions.
• a variant of the an anti-CSP antibody AB-007088 disclosed herein has a total of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,
• a variant of an anti-CSP antibody AB-007088 disclosed herein exhibits increased serum half-life as compared to AB-007088.
• a variant of an anti-CSP antibody AB-007088 disclosed herein has at least one modification, e.g., substitution, relative to the native AB-007088 Fc region of the heavy chain herein, and has improved pharmacokinetics properties, e.g., half-life.
• an Fc region of the heavy chain of such a variant of an anti-CSP antibody AB-007088 disclosed herein has at least two, three, four, five, or six, or more modifications, e.g. , substitutions.
• a variant of the an anti-CSP antibody AB-007088 disclosed herein has a total of 1, 2, 3, 4, 5, 6, 7,
• an Fc region of the heavy chain of a variant of an anti-CSP antibody AB-007088 disclosed herein includes a leucine at position 428 and a serine at position 434, wherein the numbering is defined by EU index as in Kabat.
• a variant of an anti-CSP antibody includes one, two, or three CDRs of a VL sequence of Table 25.
• a variant of an anti-CSP antibody includes at least one mutation, e.g., a substitution, and no more than 10, 20, 30, 40, or 50 mutations in the VL amino acid sequence set forth in SEQ ID NO: 217.
• the VL of the variant of an anti-CSP antibody includes at least one, at least two, or at least three mutations, e.g., substitutions, in the CDR1 amino acid sequence set forth in SEQ ID NO: 205.
• the VL of the variant of an anti-CSP antibody includes at least one, at least two, or at least three mutations, e.g., substitutions, in the framework of the amino acid sequence set forth in SEQ ID NO: 217.
• An exemplary nucleic acid sequence of SEQ ID NO: 217 is set forth in SEQ ID NO: 219.
• the VH of the variant of an anti-CSP antibody includes at least one, at least two, or at least three mutations, e.g., substitutions, in the CDR3 amino acid sequence set forth in SEQ ID NO: 210. In certain embodiments, the VH of the variant of an anti-CSP antibody includes at least one, at least two, or at least three mutations, e.g., substitutions, in the CDR1 amino acid sequence set forth in SEQ ID NO: 214. In certain embodiments, the VH of the variant of an anti-CSP antibody includes at least one, at least two, or at least three mutations, e.g., substitutions, in the CDR2 amino acid sequence set forth in SEQ ID NO: 215.
• the VH of the variant of an anti-CSP antibody includes at least one, at least two, or at least three mutations, e.g., substitutions, in the CDR3 amino acid sequence set forth in SEQ ID NO: 216.
• the mutation is a conservative substitution.
• the VH of the variant of an anti-CSP antibody includes at least one, at least two, at least three mutations, at least four mutations, at least five mutations, or at least six mutations, e.g., substitutions, in the framework of the amino acid sequence set forth in SEQ ID NO: 218.
• An exemplary nucleic acid sequence of SEQ ID NO: 218 is set forth in SEQ ID NO: 220.
• the VH of the variant of an anti-CSP antibody includes a substitution at position 28 of the SEQ ID NO: 218. In certain embodiments, the substitution is A28T. In certain embodiments, the VH of the variant of an anti-CSP antibody includes a substitution at position 40 of the SEQ ID NO: 218. In certain embodiments, the substitution is T40A. In certain embodiments, the VH of the variant of an anti-CSP antibody includes a substitution at position 69 of the SEQ ID NO: 218. In certain embodiments, the substitution is I69T. In certain embodiments, the VH of the variant of an anti-CSP antibody includes a substitution at position 80 of the SEQ ID NO: 218. In certain embodiments, the substitution is S80Y.
• the light chain of the variant of an anti-CSP antibody has an amino acid sequence set forth in SEQ ID NO: 221.
• An exemplary nucleic acid sequence of SEQ ID NO: 221 is set forth in SEQ ID NO: 224.
• a variant of an anti-CSP antibody includes at least one mutation, e.g., substitution, and no more than 10, 20, 30, 40, or 50 mutations in the Fc region of the heavy chain amino acid sequence set forth in SEQ ID NO: 222.
• the heavy chain of the variant of an anti-CSP antibody has an amino acid sequence set forth in SEQ ID NO: 222.
• An exemplary nucleic acid sequence of SEQ ID NO: 222 is set forth in SEQ ID NO: 225.
• the anti- CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 242, as shown in Table 27. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 243, as shown in Table 27. Exemplary nucleic acid sequences of SEQ ID NOS: 237, 238, 241, 242, and 243 are provided in Table 27 below.
• the anti-CSP antibody variant comprises a light chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 267, as shown in Table 30.
• the anti-CSP antibody variant comprises a heavy chain variable region comprising a CDR1, a CDR2, and a CDR3 and having the amino acid sequence set forth in SEQ ID NO: 268, as shown in Table 30.
• the anti-CSP antibody variant comprises a light chain having the amino acid sequence set forth in SEQ ID NO: 271, as shown in Table 30.
• the anti- CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 272, as shown in Table 30. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain having the amino acid sequence set forth in SEQ ID NO: 273, as shown in Table 30. Exemplary nucleic acid sequences of SEQ ID NOS: 267, 268, 271, 272, and 273 are provided in Table 30 below.
• the present disclosure provides anti-CSP antibody variants disclosed herein including one or more amino acid substitution resulting in the alteration of a glycosylation acceptor site.
• the alteration includes the elimination of the glycosylation acceptor site.
• the alteration includes modification of a glycosylation acceptor site.
• the alteration includes insertion of a glycosylation acceptor site.
• the present disclosure provides a recombinant anti circumsporozoite (CSP) antibody comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL comprises the amino acid sequence set forth in SEQ ID NO: 63 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 64.
• the recombinant antibody comprises a light chain (LC) and a heavy chain (HC), wherein the LC comprises the amino acid sequence set forth in SEQ ID NO: 67 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 69.
• RTS,S consists of two polypeptides; RTS is a single polypeptide chain corresponding to amino acids 207 to 395 of P. falciparum (3D7) that is fused to HBsAg and S is a polypeptide of 226 amino acids that corresponds to HBsAg. Stoute, et. al., N Engl J Med; 336:86-91(1997); RTS,S Clinical Trials Partnership, PLoS Med.l l(7):el001685, (2014), WO1993/10152. The RTS,S vaccine was administered with the adjuvant AS01B to increase efficacy.
• AS01B is a liposome-based formulation that contains the immunostimulants monophosphoryl lipid A (MPL) and QS21 and was shown to be more immunogenic than another adjuvant, AS02A, used in initial studies.
• MPL monophosphoryl lipid A
• AS02A another adjuvant
• All study participants were vaccinated with one of two vaccine schedules (standard full-dose: 0,1,2 M or fractional-third dose: 0,1,7 M), or placebo and subsequently challenged with a controlled human malaria parasite infection.
• Heavy and light chain AB-000224 sequences were expressed as a human IgGl monoclonal antibody.
• the present example provides the design of improved variants of AB-000224 or AB- 007088.
• the variants generated have improved developability, e.g., as identified through various in vitro assays, such as aggregation assessment by HPLC or UPLC, hydrophobic interaction chromatography (HIC), polyspecificity assays (e.g, baculovirus particle binding), self-interaction nanoparticle spectroscopy (SINS), or mass spec analysis after incubation in an accelerated degradation condition such as high temperature, low pH, high pH, or oxidative H2O2. Mutations are successful if the activity is maintained (or enhanced) while removing or reducing the severity of the liability.
• HIC hydrophobic interaction chromatography
• polyspecificity assays e.g, baculovirus particle binding
• SINS self-interaction nanoparticle spectroscopy
• mass spec analysis after incubation in an accelerated degradation condition such as high temperature, low pH, high pH, or oxidative H2O2.
• the risk of making the mutation was assessed based on: (1) the change in charge, if any, since change in charge is intrinsically risky, and a change to more positive charge is particularly risky given the already net positive charge of AB-000224 Fv; (2) conservation of the native AB- 000224 residue in the lineage versus the presence of the germline residue or other mutations at that position in the lineage and (3) the structural location of the position with respect to the NANP motif.
• Some mutations were noted to be coupled to at least one other mutation, meaning that the risk prediction is based on making the mutation in conjunction with the other mutation(s).
• Proposed AB-000224 Residue Modifications according to the ASN numbering system are shown in Table 31 below:
• each antibody was either directly coupled to a Carterra Chip or coupled using a goat anti-human Fc antibody.
• the uncoupled antibodies were washed off and various concentration gradients of the targets were flowed over the antibodies, where the highest concentration of each target was in the range 0.5-8 pg/mL
• Each antibody was immobilized in two different locations on the chip to allow for duplicate measurements.
• the affinity for each antibody-target combination was determined using 4-5 target concentrations in Mathematica software. If the variation between the two duplicates was >3 -fold, the antibody -target measurement was repeated.
• the assays were designed with opposite orientations of the target and antibody. Specifically, the target was immobilized while the antibody flowed over it in the BLI assay, while the SPR assay was designed so that the antibody was immobilized and the target flowed over it. Given these orientations, an antibody, when evaluated in the BLI assay, would be more likely to engage in binding interactions that involve multiple target molecules. As such, the binding of antibodies to targets in the BLI assay may exhibit more similarities to binding the complete CSP protein, which coats the surface of the malaria sporozoite. In contrast, the activity measured in the SPR assay would more accurately represent an interaction between an antibody F(ab) and a single target molecule.
• Table 36 The data generated for the antibodies AB-000224 and AB-007088 are summarized in Table 36 below:
• mice For each antibody, five C57B1/6 mice per experimental or control arm were administered 100 pg of antibody 16 hours prior to intravenous infection with fluorescent chimeric P. berghei sporozoites expressing P. falciparum CSP protein. Forty-two (42) hours following parasite challenge, the sporozoite liver load was quantified by bioluminescence. For each experimental mouse, the percent liver burden was calculated by subtracting the average background luminescence measured from two untreated, naive mice and calculating the percent reduction as compared to the average luminescence measured in five untreated, infected mice. The average percent reduction was reported for each of the experimental antibody groups.
• mice administered AB-000224, AB-007088 or positive control antibody AB-000317 all exhibited a similar reduction in liver burden load as compared to naive infected mice and mice treated with the negative control, AB-001245 ( Figure 1 A).
• the level of experimental and control human antibodies circulating in the mice at the time of infection was quantified via an ELISA assay ( Figure IB) and was similar between the experimental and control antibodies.
• Variants of AB-000224 and AB-007088 drove reduction in liver burden load in treated mice that was comparable to the respective parent antibody AB-000224 or AB-007088 and the positive control antibody AB-000317.
• Liver burden levels measured in animals administered either the variants, the parent molecules or positive control antibody AB-000317 was significantly lower than liver burden levels in either the naive infected mice or mice treated with the negative control, AB-001245 ( Figures 10A-10F).
• Biological replicates of each antibody variant are shown in Figures 10A-10F.
• the concentration of human antibodies that were circulating in the mice at the time of infection was determined by ELISA and is shown in Figures 10G-10I.
• C57B1/6 Mice were administered 150 pg of antibody and 16 hours later were exposed to six or seven chimeric P. berghei-infected mosquitoes. At least 70%-80% of the mosquito population are infected with chimeric P. berghei expressing P. falciparum CSP protein , resulting in exposure to at least one infectious event. Each mouse was subsequently evaluated for blood stage parasitemia from days 4-10 following infection by microscopy. Parasitemia data were recorded as positive or negative and the data used to generate a survival curve. Experimental antibodies were compared to both the positive (AB-000317) and negative (AB-001245) controls.
• mice treated with AB-000224, AB-007088 or the positive control AB-000317 were less likely to develop parasitemia as compared to mice treated with the negative control AB-001245.
• Figures 2A and 2C show AB-000224 experimental data and Figures 3 A and 3C show AB-007088 experimental data).
• the level of human antibodies circulating in the mice at the time of infection was quantified via an ELISA assay ( Figure 2B and 3B) and was similar between the experimental and control antibodies.
• mice administered variants of AB-000224 or AB-007088 were comparably likely to remain parasite-free as mice treated with either of the parent antibodies, or the positive control antibody AB-000317 ( Figures 11A -11C).
• the concentration of human antibodies that were circulating in the mice at the time of infection was determined by ELISA and is shown in Figures 11D-11F.
• AB-000224 and variants thereof showed comparable harvest yield and titer results. Thus, additional end-points were assessed.
• AB-000224 and variants thereof were evaluated for their thermal stability by differential scanning fluorimetry (DSF). Thermal unfolding was monitored by measuring the intensity of an extrinsic dye (sypro orange) as the sample was heated from 20°C to 90°C. The data was reported as Tml and Tm2 with the first transition correlating with the CH2 domain and the 2nd transition correlating with the unfolding of the Fab and CH3 domain regions. Higher unfolding temperatures are desirable and have been linked with an increase in a product’s conformational stability.
• Tm2 Lack of a Tm2 is indicative of the Fab unfolding at the same or similar temperature to the CH2 domain, reported as Tml. Additional information is also obtained from a proprietary parameter termed the weighted shoulder score which accounts for multiple pieces of information from the unfolding curve. Again, higher values are indicative of greater conformational stability.
• the DSF analysis was conducted in PBS buffer, with all samples being diluted down to a final antibody concentration of 0.15 mg/mL.
• AB-000224 and variants thereof were tested for potential hydrophobic interactions that might results in manufacturability challenges by monitoring retention time on a Zenix HPLC column. Undiluted samples were loaded onto the Zenix column and eluted isocratically with a 100 mM sodium phosphate, pH 7.0 running buffer and monitored at 220nm. Longer retention times indicates hydrophobic interactions. Most antibodies have a retention time of 8.5 - 9.0 minutes under conditions tested. AB-00224 and all variants thereof showed comparable retention times of approximately 10 minutes.
• Solubility of AB-000224 and variants thereof was assayed by precipitating the antibody samples with increasing amounts of PEG 10,000, filtering the samples, and measuring the soluble protein concentration. For each parent molecule, an initial experiment was performed to determine the ideal PEG concentration to precipitate the parent by approximately 50%, subsequent experiments used this PEG concentration on all variants to assess if a variant is more or less soluble than the parent molecule. Antibodies that precipitate above 8-10% PEG are considered highly soluble while poorly soluble antibodies precipitate at 4-5% PEG. AB-000224 and all variants thereof showed high solubility.
• the AB-007088.005 variant showed improved thermal stability by the DFS method and a T2 that approached 75 °C, while the other variants were similar to the parent AB-007088 with T1 values of approximately 70 °C and no measurable T2.
• AB-007088.005 showed a slightly reduced precipitation as compared to the parent AB-007088 and the other variants.
• AB-007088 and all variants thereof did not show low pH instability (at pH 3.3).
• AB-007088 and all variants thereof showed stability against chemical unfolding with an inflection point above 2 M guanidine (Gdn).
• AB- 007088.001 showed improved stability as compared to AB-007088 with an inflection point above 2.3 M guanidine.
• AB-007088 and variants thereof were also evaluated for their colloidal stability. When protein-protein interactions were monitored (SINS), all variants showed very low SINS values, indicating absence of protein-protein interactions. In addition, AB-007088 and variants thereof showed comparable retention times on Zenix column. Notably, when PEG solubility was determined, AB-007088 and all variants thereof showed high solubility. Further, while most of the AB-007088 variants did not show any polyreactivity, some variants showed polyreactive signal against insulin (AB-007088.001, AB-007088.003, AB-007088.004).
• the AB-007088 and variants thereof were ranked based on the different assays and titer results, and ranked as described in Figures 13B and 13C.

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