Anti-CSP Antibody

Abstract

The present disclosure provides anti-circumsporozoite protein (CSP) antibodies, compositions comprising such antibodies, as well as methods for producing the antibodies of the present disclosure and methods for treating or preventing malaria using the same.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 211,820, filed on June 17, 2021, the contents of which are incorporated by reference in their entirety.

[0002] Sequence Listing This application contains a Sequence Listing that has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. The said ASCII copy, created on Jun. 16, 2022, is named 087996_0106_SL.txt and is 461,438 bytes in size.

[0003] Field The present disclosure relates to compositions for treating or preventing malaria, and to antibodies that confer protection against infection by malaria parasites, such as Plasmodium falciparum, via insect-borne transmission. The present disclosure also relates to methods for treating, preventing, or diagnosing Plasmodium infection in a mammal. [Background technology]

[0004] background Malaria causes a significant burden of morbidity and mortality, especially in developing countries. The causative agent of malaria is a protozoan parasite, which is transmitted by mosquitoes. Several infectious Plasmodium species cause malaria, of which the most lethal is Plasmodium falciparum. Others include P. vivax, P. ovale, and P. malariae. The first generation vaccine (RTS,S) was developed using a portion of the malaria protein CSP, which contains part of the NANP repeats. CSP-based vaccines have consistently shown 30–50% efficacy in preventing erythrocytic stage infection. This level of efficacy is insufficient for eradication, and better efficacy is required for new pre-erythrocytic stage treatments. Despite the existence of antimalarial products such as mefloquinine, doxycycline, and atovaquone / proguanil, new antimalarial options are needed for cases resistant to existing antimalarials. Summary of the Invention

[0005] summary The present disclosure provides an antibody that targets Plasmodium falciparum. In certain non-limiting embodiments, the antibody is an anti-circumsporozoite protein (CSP) antibody. In certain embodiments, the recombinant antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), where 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.

[0006] In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 163; the amino acid sequence set forth in SEQ ID NO: 23; the amino acid sequence set forth in SEQ ID NO: 33; the amino acid sequence set forth in SEQ ID NO: 43; the amino acid sequence set forth in SEQ ID NO: 53; the amino acid sequence set forth in SEQ ID NO: 63; the amino acid sequence set forth in SEQ ID NO: 73; the amino acid sequence set forth in SEQ ID NO: 83; the amino acid sequence set forth in SEQ ID NO: 93; the amino acid sequence set forth in SEQ ID NO: 103; the amino acid sequence set forth in SEQ ID NO: 113; the amino acid sequence set forth in SEQ ID NO: 123; the amino acid sequence set forth in SEQ ID NO: 133; the amino acid sequence set forth in SEQ ID NO: 143; the amino acid sequence set forth in SEQ ID NO: 153; or the amino acid sequence set forth in SEQ ID NO: 173. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 164; the amino acid sequence set forth in SEQ ID NO: 24; the amino acid sequence set forth in SEQ ID NO: 34; the amino acid sequence set forth in SEQ ID NO: 44; the amino acid sequence set forth in SEQ ID NO: 54; the amino acid sequence set forth in SEQ ID NO: 64; the amino acid sequence set forth in SEQ ID NO: 74; the amino acid sequence set forth in SEQ ID NO: 84; the amino acid sequence set forth in SEQ ID NO: 94; the amino acid sequence set forth in SEQ ID NO: 104; the amino acid sequence set forth in SEQ ID NO: 114; the amino acid sequence set forth in SEQ ID NO: 124; the amino acid sequence set forth in SEQ ID NO: 134; the amino acid sequence set forth in SEQ ID NO: 144; the amino acid sequence set forth in SEQ ID NO: 154; or the amino acid sequence set forth in SEQ ID NO: 174.

[0007] In certain embodiments, 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 amino acid sequence set forth in SEQ ID NO: 84; the VL comprises the amino acid sequence set forth in SEQ ID NO: 93 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 94; or H comprises the amino acid sequence set forth in SEQ ID NO:94; or VL comprises the amino acid sequence set forth in SEQ ID NO:103 and VH comprises the amino acid sequence set forth in SEQ ID NO:104; or VL comprises the amino acid sequence set forth in SEQ ID NO:113 and VH comprises the amino acid sequence set forth in SEQ ID NO:114; or VL comprises the amino acid sequence set forth in SEQ ID NO:123 and VH comprises the amino acid sequence set forth in SEQ ID NO:124; or VL comprises the amino acid sequence set forth in SEQ ID NO:133 and VH comprises the amino acid sequence set forth in SEQ ID NO:134; or VL comprises the amino acid sequence set forth in SEQ ID NO:143 and VH comprises the amino acid sequence set forth in SEQ ID NO:144; or VL comprises the amino acid sequence set forth in SEQ ID NO:153 and VH comprises the amino acid sequence set forth in SEQ ID NO:154; or VL comprises the amino acid sequence set forth in SEQ ID NO:173 and VH comprises the amino acid sequence set forth in SEQ ID NO:174.

[0008] In certain embodiments, 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. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 133 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 134. In certain embodiments, 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.

[0009] In certain embodiments, the recombinant antibody comprises a light chain (LC) and a heavy chain (HC). In certain embodiments, 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:167; or the amino acid sequence set forth in SEQ ID NO:177. In certain embodiments, the HC comprises the amino acid sequence set forth in SEQ ID NO:28 or SEQ ID NO:29; the amino acid sequence set forth in SEQ ID NO:38 or SEQ ID NO:39; the amino acid sequence set forth in SEQ ID NO:48 or SEQ ID NO:49; the amino acid sequence set forth in SEQ ID NO:58 or SEQ ID NO:59; the amino acid sequence set forth in SEQ ID NO:68 or SEQ ID NO:69; the amino acid sequence set forth in SEQ ID NO:78 or SEQ ID NO:79; the amino acid sequence set forth in SEQ ID NO:88 or SEQ ID NO:89; the amino acid sequence set forth in SEQ ID NO:98 or SEQ ID NO:99; the amino acid sequence set forth in SEQ ID NO:108 or SEQ ID NO:109; the amino acid sequence set forth in SEQ ID NO:118 or SEQ ID NO:119; the amino acid sequence set forth in SEQ ID NO:128 or SEQ ID NO:129; the amino acid sequence set forth in SEQ ID NO:138 or SEQ ID NO:139; the amino acid sequence set forth in SEQ ID NO:148 or SEQ ID NO:149; the amino acid sequence set forth in SEQ ID NO:158 or SEQ ID NO:159; the amino acid sequence set forth in SEQ ID NO:168 or SEQ ID NO:169; or the amino acid sequence set forth in SEQ ID NO:178 or SEQ ID NO:179.

[0010] In certain embodiments, the LC comprises the amino acid sequence set forth in SEQ ID NO:27 and the HC comprises the amino acid sequence set forth in SEQ ID NO:28 or SEQ ID NO:29; the LC comprises the amino acid sequence set forth in SEQ ID NO:37 and the HC comprises the amino acid sequence set forth in SEQ ID NO:38 or SEQ ID NO:39; the LC comprises the amino acid sequence set forth in SEQ ID NO:47 and the HC comprises the amino acid sequence set forth in SEQ ID NO:48 or SEQ ID NO:49; the LC comprises the amino acid sequence set forth in SEQ ID NO:57 and the HC comprises the amino acid sequence set forth in SEQ ID NO:58 or SEQ ID NO:59; 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:68 or SEQ ID NO:69; the LC comprises the amino acid sequence set forth in SEQ ID NO:77 and the HC comprises the amino acid sequence set forth in SEQ ID NO:78 or SEQ ID NO:79; the LC comprises the amino acid sequence set forth in SEQ ID NO:87 and the HC comprises the amino acid sequence set forth in SEQ ID NO:88 or SEQ ID NO:89; the LC comprises the amino acid sequence set forth in SEQ ID NO:97 and the HC comprises the amino acid sequence set forth in SEQ ID NO:98 or SEQ ID NO:99; the LC comprises the amino acid sequence set forth in SEQ ID NO: or LC comprises the amino acid sequence set forth in SEQ ID NO:107, and HC comprises the amino acid sequence set forth in SEQ ID NO:108 or SEQ ID NO:109; LC comprises the amino acid sequence set forth in SEQ ID NO:117, and HC comprises the amino acid sequence set forth in SEQ ID NO:118 or SEQ ID NO:119; LC comprises the amino acid sequence set forth in SEQ ID NO:127, and HC comprises the amino acid sequence set forth in SEQ ID NO:128 or SEQ ID NO:129; LC comprises the amino acid sequence set forth in SEQ ID NO:137, and HC comprises the amino acid sequence set forth in SEQ ID NO:138 or SEQ ID NO:139; LC comprises the amino acid sequence set forth in SEQ ID NO:147, and HC comprises the amino acid sequence set forth in SEQ ID NO:148 or SEQ ID NO:149; LC comprises the amino acid sequence set forth in SEQ ID NO:157, and HC comprises the amino acid sequence set forth in SEQ ID NO:158 or SEQ ID NO:159; LC comprises the amino acid sequence set forth in SEQ ID NO:167, and HC comprises the amino acid sequence set forth in SEQ ID NO:168 or SEQ ID NO:169; or LC comprises the amino acid sequence set forth in SEQ ID NO:177, and HC comprises the amino acid sequence set forth in SEQ ID NO:178 or SEQ ID NO:179.

[0011] In certain embodiments, 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. In certain embodiments, the LC comprises the amino acid sequence set forth in SEQ ID NO: 137 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 139. In certain embodiments, the LC comprises the amino acid sequence set forth in SEQ ID NO: 167 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 169.

[0012] In certain embodiments, the recombinant antibody comprises 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: 13. In certain embodiments, the VL comprises at least one amino acid substitution. In certain embodiments, the at least one amino acid substitution is at position 1 and / or position 44. In certain embodiments, the amino acid substitution at position 1 is E1Q. In certain embodiments, the amino acid substitution at position 44 is R44T. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 14. In certain embodiments, the VH comprises at least one amino acid substitution. In certain embodiments, the at least one amino acid substitution is at position 21, 23, 88, 98, or a combination thereof. In certain embodiments, the amino acid substitution at position 1 is E1Q. In certain embodiments, the amino acid substitution at position 44 is R44T. In certain embodiments, the amino acid substitution at position 21 is P21S. In certain embodiments, the amino acid substitution at position 23 is T23A. In certain embodiments, the amino acid substitution at position 80 is I80T. In certain embodiments, the amino acid substitution at position 90 is T90A.

[0013] In certain embodiments, the recombinant antibody comprises a heavy chain (HC) comprising the amino acid sequence set forth in SEQ ID NO: 18. In certain embodiments, the HC comprises at least one amino acid substitution. In certain embodiments, the at least one amino acid substitution is at position 438 and / or 444. In certain embodiments, the amino acid substitution at position 438 is M438L. In certain embodiments, the amino acid substitution at position 444 is N444S.

[0014] In certain non-limiting embodiments, the present disclosure also provides a recombinant anti-circumsporozoite protein (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: 163 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 164. In certain embodiments, 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: 167 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 169.

[0015] In certain non-limiting embodiments, the present disclosure provides a recombinant anti-circumsporozoite protein (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. In certain embodiments, 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.

[0016] In certain non-limiting embodiments, the disclosure further provides a recombinant anti-circumsporozoite protein (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: 133 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 134. In certain embodiments, 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: 137 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 139.

[0017] In certain non-limiting embodiments, the present disclosure provides a recombinant anti-circumsporozoite protein (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: 183, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 184, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 185; and the heavy chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 186, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 187, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 188. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 195, and the VH comprises the amino acid sequence set forth in SEQ ID NO: 196. In certain embodiments, the antibody comprises a LC and a HC. In certain embodiments, the LC comprises the amino acid sequence set forth in SEQ ID NO: 199, and the HC comprises the amino acid sequence set forth in SEQ ID NO: 200 or SEQ ID NO: 201.

[0018] In certain non-limiting embodiments, the present disclosure provides a recombinant anti-circumsporozoite protein (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.

[0019] In certain embodiments, the amino acid sequence set forth in SEQ ID NO: 227; the amino acid sequence set forth in SEQ ID NO: 237; the amino acid sequence set forth in SEQ ID NO: 247; the amino acid sequence set forth in SEQ ID NO: 257; or the amino acid sequence set forth in SEQ ID NO: 267. In certain embodiments, the VH comprises the amino acid sequence set forth in SEQ ID NO: 228; the amino acid sequence set forth in SEQ ID NO: 238; the amino acid sequence set forth in SEQ ID NO: 248; the amino acid sequence set forth in SEQ ID NO: 258; or the amino acid sequence set forth in SEQ ID NO: 268.

[0020] In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 227 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 228; the VL comprises the amino acid sequence set forth in SEQ ID NO: 237 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 238; the VL comprises the amino acid sequence set forth in SEQ ID NO: 247 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 248; the VL comprises the amino acid sequence set forth in SEQ ID NO: 257 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 258; or the VL comprises the amino acid sequence set forth in SEQ ID NO: 267 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 268.

[0021] In certain embodiments, the recombinant antibody comprises an LC and an HC. In certain embodiments, the LC comprises the amino acid sequence set forth in SEQ ID NO:231; the amino acid sequence set forth in SEQ ID NO:241; the amino acid sequence set forth in SEQ ID NO:251; the amino acid sequence set forth in SEQ ID NO:261; or the amino acid sequence set forth in SEQ ID NO:271. In certain embodiments, the HC comprises the amino acid sequence set forth in SEQ ID NO:232 or SEQ ID NO:233; the amino acid sequence set forth in SEQ ID NO:242 or SEQ ID NO:243; the amino acid sequence set forth in SEQ ID NO:252 or SEQ ID NO:253; the amino acid sequence set forth in SEQ ID NO:262 or SEQ ID NO:263; or the amino acid sequence set forth in SEQ ID NO:272 or SEQ ID NO:273.

[0022] In certain embodiments, the LC comprises the amino acid sequence set forth in SEQ ID NO:231 and the HC comprises the amino acid sequence set forth in SEQ ID NO:232 or SEQ ID NO:233; the LC comprises the amino acid sequence set forth in SEQ ID NO:241 and the HC comprises the amino acid sequence set forth in SEQ ID NO:242 or SEQ ID NO:243; the LC comprises the amino acid sequence set forth in SEQ ID NO:251 and the HC comprises the amino acid sequence set forth in SEQ ID NO:252 or SEQ ID NO:253; the LC comprises the amino acid sequence set forth in SEQ ID NO:261 and the HC comprises the amino acid sequence set forth in SEQ ID NO:262 or SEQ ID NO:263; or the LC comprises the amino acid sequence set forth in SEQ ID NO:271 and the HC comprises the amino acid sequence set forth in SEQ ID NO:272 or SEQ ID NO:273.

[0023] In certain embodiments, the VH comprises an amino acid sequence set forth in SEQ ID NO: 218. In certain embodiments, the VH comprises at least one amino acid substitution. In certain embodiments, the at least one amino acid substitution is at position 40, 69, 80, 85, 120, or a combination thereof. In certain embodiments, the amino acid substitution at position 40 is T40A. In certain embodiments, the amino acid substitution at position 69 is I69T. In certain embodiments, the amino acid substitution at position 80 is S80Y. In certain embodiments, the amino acid substitution at position 85 is G85S. In certain embodiments, the amino acid substitution at position 120 is I120T. In certain embodiments, the HC comprises an amino acid sequence set forth in SEQ ID NO: 222. In certain embodiments, the HC comprises at least one amino acid substitution. In certain embodiments, the at least one amino acid substitution is at position 434 and / or 440. In certain embodiments, the amino acid substitution at position 434 is M434L. In certain embodiments, the amino acid substitution at position 440 is N440S.

[0024] In certain embodiments, the recombinant antibody exhibits at least a 20% reduction in parasite liver burden compared to the reference antibody. In certain embodiments, the recombinant antibody exhibits at least a 20% increase in viability compared to the reference antibody. In certain embodiments, the recombinant antibody exhibits increased conformational stability compared to the reference antibody. In certain embodiments, the recombinant antibody exhibits increased colloidal stability compared to the reference antibody. In certain embodiments, the reference antibody is AB-000317. In certain embodiments, the reference antibody is AB-000224. In certain embodiments, the reference antibody is AB-007088.

[0025] In certain embodiments, the recombinant antibody binds to the 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.

[0026] In certain embodiments, the recombinant antibody comprises at least one modification 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 to the native AB-000224 variable light chain amino acid sequence set forth in SEQ ID NO: 13. In certain embodiments, the recombinant antibody comprises at least one modification to the native AB-000224 variable heavy chain amino acid sequence set forth in SEQ ID NO: 14 and at least one modification to the native AB-000224 variable light chain amino acid sequence set forth in SEQ ID NO: 13.

[0027] In certain embodiments, the recombinant antibody comprises at least one modification to the native AB-007088 variable heavy chain amino acid sequence set forth in SEQ ID NO: 218. In certain embodiments, the recombinant antibody comprises at least one modification to the native AB-007088 variable light chain amino acid sequence set forth in SEQ ID NO: 217. In certain embodiments, the recombinant antibody comprises at least one modification to the native AB-007088 variable heavy chain amino acid sequence set forth in SEQ ID NO: 218 and at least one modification to the native AB-000224 variable light chain amino acid sequence set forth in SEQ ID NO: 217.

[0028] In certain non-limiting embodiments, the disclosure provides a polynucleotide encoding an antibody disclosed herein. In certain non-limiting embodiments, the disclosure provides an expression vector comprising a polynucleotide disclosed herein. In certain non-limiting embodiments, the disclosure provides a host cell comprising an expression vector or a polynucleotide disclosed herein.

[0029] In certain non-limiting embodiments, the present disclosure provides a composition comprising an antibody disclosed herein. In certain embodiments, the composition further comprises a pharma- ceutically acceptable carrier.

[0030] In certain non-limiting embodiments, the present disclosure provides a method of preventing and / or treating malaria in a subject in need thereof comprising administering an effective amount of an antibody disclosed herein or a composition disclosed herein. In certain embodiments, the subject is a pediatric patient.

[0031] In certain non-limiting embodiments, the present disclosure provides an antibody or composition disclosed herein for use in the prophylactic and / or therapeutic treatment of malaria in a subject in need of treatment. Additionally, in certain non-limiting embodiments, the present disclosure provides an antibody or composition disclosed herein for the manufacture of a medicament for the prophylactic and / or therapeutic treatment of malaria in a subject in need of treatment. Furthermore, the present disclosure provides the use of an antibody or composition disclosed herein for the prophylactic and / or therapeutic treatment and / or prophylaxis of malaria in a subject in need of treatment. In certain embodiments, the subject is a pediatric patient.

[0032] BRIEF DESCRIPTION OF THE DRAWINGS [Brief description of the drawings]

[0033] [Figure 1A] Figure 1A shows the parasite liver burden after administration of experimental anti-CSP antibodies AB-000224 and AB-007088. Parasite liver burden was measured by bioluminescence (photons / second) generated from fluorescent sporozoites (y-axis). Each point indicates the total amount of bioluminescence measured in one mouse and thus the liver burden of sporozoites. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 1B] Figure 1B shows the serum concentration of relevant human IgG measured by ELISA 15 hours after antibody administration. Each point indicates the total amount of human IgG in one mouse. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 2A]Figures 2A and 2C show survival rates in two experiments in mice treated with the anti-CSP antibody AB-000224 and exposed to mosquitoes infected with chimeric P. berghei expressing the P. falciparum CSP protein. [Figure 2B] Figure 2B shows the relevant human IgG serum concentrations measured by ELISA obtained 15 hours after antibody administration in the experiment shown in Figure 2A. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 2C] Figures 2A and 2C show survival rates in two experiments in mice treated with the anti-CSP antibody AB-000224 and exposed to mosquitoes infected with chimeric P. berghei expressing the P. falciparum CSP protein. [Figure 3A] Figures 3A and 3C show survival rates in two experiments in mice treated with the anti-CSP antibody AB-007088 and exposed to mosquitoes infected with chimeric P. berghei expressing the P. falciparum CSP protein. [Figure 3B] Figure 3B shows the relevant human IgG serum concentrations measured by ELISA obtained 15 hours after antibody administration in the experiment shown in Figure 3A. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 3C] Figures 3A and 3C show survival rates in two experiments in mice treated with the anti-CSP antibody AB-007088 and exposed to mosquitoes infected with chimeric P. berghei expressing the P. falciparum CSP protein. [Figure 4A] Figures 4A-4B show the sequences of the AB-000224 antibody. Figure 4A shows the lambda light chain sequence of AB-000224 (SEQ ID NO: 286). Figure 4B shows the IgG1 heavy chain sequence of AB-000224 (SEQ ID NO: 287). The framework and CDR regions are designated using the ASN system. [Figure 4B] Figures 4A-4B show the sequences of the AB-000224 antibody. Figure 4A shows the lambda light chain sequence of AB-000224 (SEQ ID NO: 286). Figure 4B shows the IgG1 heavy chain sequence of AB-000224 (SEQ ID NO: 287). The framework and CDR regions are designated using the ASN system. [Figure 5A] Figures 5A-5B show the sequences of the AB-007088 antibody. Figure 5A shows the lambda light chain sequence of AB-007088 (SEQ ID NO: 288). Figure 5B shows the IgG1 heavy chain sequence of AB-007088 (SEQ ID NO: 289). The framework and CDR regions are designated using the ASN system. [Figure 5B] Figures 5A-5B show the sequences of the AB-007088 antibody. Figure 5A shows the lambda light chain sequence of AB-007088 (SEQ ID NO: 288). Figure 5B shows the IgG1 heavy chain sequence of AB-007088 (SEQ ID NO: 289). The framework and CDR regions are designated using the ASN system. [Figure 6] FIG. 6 shows the ASN numbering system for the light chain of AB-000224. [Figure 7] FIG. 7 shows the ASN numbering system of the heavy chain of AB-000224. [Figure 8] FIG. 8 shows the ASN numbering system for the light chain of AB-007088. [Figure 9] FIG. 9 shows the ASN numbering system for the heavy chain of AB-007088. [Figure 10A]Figures 10A-10F show parasite liver burden following administration of experimental anti-CSP antibody variants disclosed herein. Results are expressed as percent inhibition, with naive infection set at 100%. Figures 10G-10I show concentrations of human antibodies circulating in mice at the time of infection, measured by ELISA for the assays shown in Figures 10D-F, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 10B] Figures 10A-10F show parasite liver burden following administration of experimental anti-CSP antibody variants disclosed herein. Results are expressed as percent inhibition, with naive infection set at 100%. Figures 10G-10I show concentrations of human antibodies circulating in mice at the time of infection, measured by ELISA for the assays shown in Figures 10D-F, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 10C] Figures 10A-10F show parasite liver burden following administration of experimental anti-CSP antibody variants disclosed herein. Results are expressed as percent inhibition, with naive infection set at 100%. Figures 10G-10I show concentrations of human antibodies circulating in mice at the time of infection, measured by ELISA for the assays shown in Figures 10D-F, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 10D]Figures 10A-10F show parasite liver burden following administration of experimental anti-CSP antibody variants disclosed herein. Results are expressed as percent inhibition, with naive infection set at 100%. Figures 10G-10I show concentrations of human antibodies circulating in mice at the time of infection, measured by ELISA for the assays shown in Figures 10D-F, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 10E] Figures 10A-10F show parasite liver burden following administration of experimental anti-CSP antibody variants disclosed herein. Results are expressed as percent inhibition, with naive infection set at 100%. Figures 10G-10I show concentrations of human antibodies circulating in mice at the time of infection, measured by ELISA for the assays shown in Figures 10D-F, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 10F] Figures 10A-10F show parasite liver burden following administration of experimental anti-CSP antibody variants disclosed herein. Results are expressed as percent inhibition, with naive infection set at 100%. Figures 10G-10I show concentrations of human antibodies circulating in mice at the time of infection, measured by ELISA for the assays shown in Figures 10D-F, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 10G]Figures 10A-10F show parasite liver burden following administration of experimental anti-CSP antibody variants disclosed herein. Results are expressed as percent inhibition, with naive infection set at 100%. Figures 10G-10I show concentrations of human antibodies circulating in mice at the time of infection, measured by ELISA for the assays shown in Figures 10D-F, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 10H] Figures 10A-10F show parasite liver burden following administration of experimental anti-CSP antibody variants disclosed herein. Results are expressed as percent inhibition, with naive infection set at 100%. Figures 10G-10I show concentrations of human antibodies circulating in mice at the time of infection, measured by ELISA for the assays shown in Figures 10D-F, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 10I] Figures 10A-10F show parasite liver burden following administration of experimental anti-CSP antibody variants disclosed herein. Results are expressed as percent inhibition, with naive infection set at 100%. Figures 10G-10I show concentrations of human antibodies circulating in mice at the time of infection, measured by ELISA for the assays shown in Figures 10D-F, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 11A]Figures 11A-11C show survival of mice following administration of anti-CSP antibody variants disclosed herein and exposure to mosquitoes infected with chimeric P. berghei expressing the P. falciparum CSP protein. Figures 11D-F show concentrations of human antibodies circulating in mice at the time of infection as measured by ELISA for the assays shown in Figures 11A-C, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 11B] Figures 11A-11C show survival of mice following administration of anti-CSP antibody variants disclosed herein and exposure to mosquitoes infected with chimeric P. berghei expressing the P. falciparum CSP protein. Figures 11D-F show concentrations of human antibodies circulating in mice at the time of infection as measured by ELISA for the assays shown in Figures 11A-C, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 11C] Figures 11A-11C show survival of mice following administration of anti-CSP antibody variants disclosed herein and exposure to mosquitoes infected with chimeric P. berghei expressing the P. falciparum CSP protein. Figures 11D-F show concentrations of human antibodies circulating in mice at the time of infection as measured by ELISA for the assays shown in Figures 11A-C, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 11D]Figures 11A-11C show survival of mice following administration of anti-CSP antibody variants disclosed herein and exposure to mosquitoes infected with chimeric P. berghei expressing the P. falciparum CSP protein. Figures 11D-F show concentrations of human antibodies circulating in mice at the time of infection as measured by ELISA for the assays shown in Figures 11A-C, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 11E] Figures 11A-11C show survival of mice following administration of anti-CSP antibody variants disclosed herein and exposure to mosquitoes infected with chimeric P. berghei expressing the P. falciparum CSP protein. Figures 11D-F show concentrations of human antibodies circulating in mice at the time of infection as measured by ELISA for the assays shown in Figures 11A-C, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 11F] Figures 11A-11C show survival of mice following administration of anti-CSP antibody variants disclosed herein and exposure to mosquitoes infected with chimeric P. berghei expressing the P. falciparum CSP protein. Figures 11D-F show concentrations of human antibodies circulating in mice at the time of infection as measured by ELISA for the assays shown in Figures 11A-C, respectively. AB-001245 is a non-malaria specific antibody and was used as a negative control. AB-000317 is a positive control. [Figure 12A] FIG. 12A shows the biophysical characteristics of AB-000224 and its variants. [Figure 12B] FIG. 12B shows the ranking of AB-000224 and its variants based on all data obtained during the analysis of biophysical features. [Figure 12C]FIG. 12C shows the ranking including potency, excluding polyreactivity, of AB-000224 and its variants. [Figure 13A] FIG. 13A shows the biophysical characteristics of AB-007088 and its variants. [Figure 13B] FIG. 13B shows the ranking of AB-007088 and its variants based on all data obtained during the analysis of biophysical features. [Figure 13C] FIG. 13C shows the ranking including potency, excluding polyreactivity, of AB-007088 and its variants. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] Detailed Description The present disclosure is based, at least in part, on the discovery of several anti-CSP antibody variants. The present disclosure surprisingly demonstrates that the disclosed anti-CSP antibodies and variants thereof have superior properties compared to previously disclosed antibodies.

[0035] definition For purposes of interpreting this specification, the following definitions will apply, except that, where appropriate, terms used in the singular will also include the plural and vice versa. In the event that a definition set forth below conflicts with any document incorporated herein by reference, the definition set forth below shall control.

[0036] Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Singleton et al., Dictionary of Microbiology and Molecular Biology 2nd ed., J. Wiley & Sons (New York, NY 1994) and March, Advanced Organic Chemistry Reactions, Mechanisms and Structure 4th ed., John Wiley & Sons (New York, NY 1992) provide those skilled in the art with a general guide to many of the terms used in this application.

[0037] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context dictates otherwise. Thus, for example, reference to a "protein" or an "antibody" includes a plurality of proteins or antibodies respectively; reference to a "cell" includes a mixture of cells, and the like.

[0038] As used herein, the term "about" or "approximately" refers to a normal error range for the respective value, readily known to one of ordinary skill in the art, e.g., ±20%, ±10%, or ±5%, within the intended meaning of the stated value.

[0039] As used herein, the term "antibody" refers to an isolated or recombinant binding agent that contains the necessary variable region sequence to specifically bind to an antigen epitope. Thus, as used herein, "antibody" refers to any form of antibody or fragment thereof that exhibits the desired biological activity, e.g., binding to a specific target antigen. Thus, it is used in the broadest sense and specifically encompasses monoclonal antibodies (including full-length monoclonal antibodies), human antibodies, chimeric antibodies, nanobodies, diabodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, including but not limited to scFv, Fab, etc., so long as they exhibit the desired biological activity.

[0040] An "antibody fragment" includes a portion of an intact antibody, e.g., the antigen-binding or variable region of an intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, Fv fragments; diabodies; linear antibodies (e.g., Zapata et al., Protein Eng. 8(10):1057-1062(1995)); single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments. Papain digestion of an antibody produces two identical antigen-binding fragments, called "Fab" fragments, each with a single antigen-binding site, and a residual "Fc" fragment (a designation reflecting the ability to be readily crystallized). Pepsin treatment produces an F(ab')2 fragment that has two antigen-binding sites and is still capable of cross-linking antigen.

[0041] As used herein, a "recombinant antibody" refers to an antibody whose exact amino acid sequence does not naturally occur in a given organism (e.g., an antibody derived from a mammal). In certain embodiments, the term may refer to an antibody that contains one or more amino acid residues not found in a naturally occurring antibody. In certain embodiments, a recombinant antibody may have a CDR that contains amino acid residues not found in a naturally occurring antibody (e.g., an antibody derived from a mammal). In another exemplary embodiment, a recombinant antibody may have a framework (FR) that contains amino acid residues not found in a naturally occurring antibody (e.g., an antibody derived from a mammal). In certain embodiments, a recombinant antibody may have a constant region that contains amino acid residues not found in a naturally occurring antibody (e.g., an antibody derived from a mammal). In certain embodiments, a recombinant antibody is a variant of a naturally occurring antibody (e.g., AB-000224) that contains at least one modification, e.g., substitution, to the native variable heavy or light amino acid sequence. For example, but not limited to, a recombinant antibody can be the anti-CSP antibody AB-000224 disclosed herein that includes at least one modification, e.g., a substitution, relative to the native AB-000224 variable heavy chain amino acid sequence (SEQ ID NO: 14) or variable light chain amino acid sequence (SEQ ID NO: 13) described herein. Recombinant antibodies have improved developability, e.g., reduced heterogeneity, increased yield, increased stability, improved net charge for improved pharmacokinetics, and / or reduced immunogenicity.

[0042] As used herein, the terms "anti-CSP antibody" and "CSP antibody" are used synonymously and refer to an antibody that binds to the circumsporozoite protein (CSP) antigen of Plasmodium falciparum.

[0043] An "antibody that binds to the same epitope" as a reference antibody refers to an antibody that blocks the binding of the reference antibody to its antigen by 50% or more in a competitive assay, and conversely, the reference antibody blocks the binding of the antibody to its antigen by 50% or more in a competitive assay.

[0044] As used herein, "V region" refers to an antibody variable region domain comprising framework 1, CDR1, framework 2, CDR2, framework 3, CDR3, and framework 4 segments. The heavy chain V region, VH, is the result of rearrangement of the V gene (HV), D gene (HD) and J gene (HJ) during B cell differentiation in a manner known as V(D)J recombination. The light chain V region, VL, is the result of rearrangement of the V gene (LV) and J gene. In certain embodiments, the terms "VH" and "heavy chain variable" refer to the heavy chain V region of an antibody. In certain embodiments, the terms "VL" and "light chain variable" refer to the light chain V region of an antibody.

[0045] As used herein, "complementarity determining region (CDR)" refers to the three hypervariable regions (HVRs) in each chain that interrupt the four "framework" regions established by the light and heavy chain variable regions. The CDRs are the main contributors to binding to an epitope of an antigen. The CDRs of each chain are numbered sequentially from the N-terminus and are referred to as CDR1, CDR2, CDR3, and are also identified by the chain in which the particular CDR is located. Thus, a VH CDR3 (HCDR3) is located in the variable domain of the heavy chain of the antibody in which it is found, while a VL CDR3 (LCDR3) is the CDR3 of the variable domain of the light chain of the antibody in which it is found. The term "CDR" is used interchangeably with "HVR" when referring to the sequence of a CDR.

[0046] The amino acid sequences of the CDRs and framework regions can be determined using various definitions in the art, e.g., Kabat, Chothia, the international ImMunoGeneTics database (IMGT), and AbM (see, e.g., Chothia & Lesk, 1987, Canonical structures for the hypervariable regions of immunoglobulins. J. Mol. Biol. 196, 901-917; Chothia C. et al., 1989, Conformations of immunoglobulin hypervariable regions. Nature 342, 877-883; Chothia C. et al., 1992, Structural repertoire of the human VH segments J. Mol. Biol. 227, 799-817; Al-Lazikani et al., J. Mol. Biol 1997, 273(4)).Definitions of antigen binding sites are also described in: Ruiz et al., IMGT, the international ImMunoGeneTics database. Nucleic Acids Res., 28, 219-221 (2000); and Lefranc, M.-P. IMGT, the international ImMunoGeneTics database. Nucleic Acids Res. Jan 1;29(1):207-9 (2001); MacCallum et al, Antibody-antigen interactions: Contact analysis and binding site topography, J. Mol. Biol., 262(5), 732-745 (1996); and Martin et al, Proc. Natl Acad. Sci. USA, 86, 9268-9272 (1989); Martin, et al, Methods Enzymol., 203, 121-153, (1991); Pedersen et al. al, Immunomethods, 1, 126, (1992); and Rees et al, In Sternberg MJE (ed.), Protein Structure Prediction. Oxford University Press, Oxford, 141-172 1996). References to CDRs determined by Kabat numbering are based on, for example, Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institute of Health, Bethesda, MD (1991). Chothia CDRs are determined as defined by Chothia (see, for example, Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). In certain embodiments, the amino acid sequences of the CDRs and framework regions are numbered using the antibody structure numbering (ASN) system.Antibody structural numbering (ASN) is a numbering system developed based on the AHo numbering system defined by Annemarie Honegger for the variable regions (Honegger & Pluckthun, J. Mol. Honegger & Pluckthun, J. Mol. Biol. 309:657-670 (2001)) and extended to include the constant domain. Figures 6-7 show the ASN numbering of the light and heavy chains of AB-000224-LS, respectively. Figures 8-9 show the ASN numbering of the light and heavy chains of AB-007088-LS, respectively.

[0047] "Fc region" refers to the constant region of an antibody excluding the first constant region immunoglobulin domain. Thus, Fc refers to the last two constant region immunoglobulin domains of IgA, IgD and IgG, the last three constant region immunoglobulin domains of IgE and IgM, and the flexible hinge at the N-terminus of these domains. The Fc of IgA and IgM may include the J chain. For IgG, Fc includes the immunoglobulin domains Cy2 and Cy3, and the hinge between Cyl and Cy. Although it is understood in the art that the boundaries of the Fc region may vary, the human IgG heavy chain Fc region is usually defined as including residues C226 or P230 to the carboxyl terminus thereof, using numbering according to the EU index, as in Kabat et al. (1991, NIH Publication 91-3242, National Technical Information Service, Springfield, Va.). The term "Fc region" may refer to this region alone or in the context of an antibody or antibody fragment. "Fc region" includes naturally occurring allelic variants of the Fc region as well as modifications that modulate effector functions. Fc region also includes variants that do not alter biological function. For example, one or more amino acids may be deleted from the N-terminus or C-terminus of the Fc region of an immunoglobulin without substantially impairing biological function. Such variants may be selected according to general rules known in the art to minimize impact on activity (see, for example, Bowie, et al., Science 247:306-1310, 1990). For example, in an IgG4 antibody, a single amino acid substitution (S228P according to Kabat numbering; designated IgG4Pro) may be introduced to eliminate the heterogeneity observed in recombinant IgG4 antibodies (see, for example, Angal, et al., Mol Immunol 30:105-108, 1993). In certain embodiments, the Fc region comprises substitutions that improve the pharmacokinetic properties of the antibody, e.g., substitutions that increase serum half-life.Non-limiting examples of Fc region replacements can be found in US Pat. No. 8,088,376, the contents of which are incorporated by reference in their entirety.

[0048] The term "equilibrium dissociation constant," abbreviated as KD, refers to the dissociation rate constant (kd, time -1 ) to the association rate constant (ka, time -1 M -1 ) divided by the equilibrium dissociation constant. The equilibrium dissociation constant may be measured by any method. Thus, in certain embodiments, the antibodies of the present disclosure have a KD of less than about 50 nM, typically less than about 25 nM, or less than 10 nM, e.g., less than about 5 nM, or less than about 1 nM, and often less than about 10 nM, as determined by surface plasmon resonance analysis using a biosensor system such as a Biacore® system performed at 37°C. In certain embodiments, the antibodies of the present disclosure have a KD of less than 5 x 10 when measured as a bivalent antibody. -5 Less than M, 10 -5 Less than M, 5×10 -6 Less than M, 10 -6 Less than M, 5×10 -7 Less than M, 10 -7 Less than M, 5×10 -8 Less than M, 10 -8 Less than M, 5×10 -9 Less than M, 10 -9 Less than M, 5×10 -10 Less than M, 10 -10 Less than M, 5×10 -11 Less than M, 10 -11 Less than M, 5×10 -12 Less than M, 10 -12 Less than M, 5×10 -13 Less than M, 10 -13 Less than M, 5×10 -14 Less than M, 10 -14 Less than M, 5×10 -15 Less than M or 10 -15 As used herein, an "improved" KD refers to a lower KD. In certain embodiments, the antibodies of the present disclosure have a KD of 5×10 or less when measured as a monovalent antibody, e.g., a monovalent Fab. -5 Less than M, 10 -5 Less than M, 5×10-6 Less than M, 10 -6 Less than M, 5×10 -7 Less than M, 10 -7 Less than M, 5×10 -8 Less than M, 10 -8 Less than M, 5×10 -9 Less than M, 10 -9 Less than M, 5×10 -10 Less than M, 10 -10 Less than M, 5×10 -11 Less than M, 10 -11 Less than M, 5×10 -12 Less than M, 10 -12 Less than M, 5×10 -13 Less than M, 10 -13 Less than M, 5×10 -14 Less than M, 10 -14 Less than M, 5×10 -15 Less than M or 10 -15 In certain embodiments, the anti-CSP antibodies of the disclosure have a KD of less than 100 pM, or, for example, less than 75 pM, e.g., in the range of 1 to 100 pM, as measured by surface plasmon resonance analysis using a biosensor system such as a Biacore® system performed at 37° C. In certain embodiments, the anti-CSP antibodies of the disclosure have a KD of more than 100 pM, e.g., in the range of 100-1000 pM or 500-1000 pM, as measured by surface plasmon resonance analysis using a biosensor system such as a Biacore® system performed at 37° C.

[0049] The term "monovalent molecule" as used herein refers to a molecule that has one antigen-binding site, e.g., a Fab or scFv.

[0050] The term "bivalent molecule" as used herein refers to a molecule that has two antigen binding sites. In certain embodiments, the bivalent molecule of the invention is a bivalent antibody or a bivalent fragment thereof. In certain embodiments, the bivalent molecule of the invention is a bivalent antibody. In certain embodiments, the bivalent molecule of the invention is an IgG. In certain embodiments, monoclonal antibodies have a bivalent basic structure. IgG and IgE have only one bivalent unit, whereas IgA and IgM have a higher valency because they are composed of multiple bivalent units (two and five, respectively). This bivalency increases the avidity of the antibody for the antigen.

[0051] As used herein, the terms "monovalent binding" or "monovalently bind" refer to the binding of one antigen-binding site to its antigen.

[0052] As used herein, the term "bivalent binding" or "bivalently bind" refers to both antigen-binding sites of a bivalent molecule binding to the antigen. In certain embodiments, both antigen-binding sites of a bivalent molecule share the same antigen specificity.

[0053] The term "valency" as used herein refers to the number of different binding sites of an antibody for an antigen. A monovalent antibody contains one binding site for an antigen. A divalent antibody (e.g., a bivalent IgG antibody) contains two binding sites for the same antigen.

[0054] The term "affinity" as used herein refers to the strength, alone or in combination, of one or both arms of an antibody (e.g., an IgG antibody) to bind to either a simple or complex antigen expressing one or more epitopes. As defined herein, the term "affinity" does not imply a particular valency between two binding partners.

[0055] The phrases "specifically (or selectively) bind" or "specifically (or selectively) immunoreact with" an antigen or target, when referring to a protein or peptide, refer to a binding reaction in which an antibody binds to the antigen or target of interest with an affinity that is distinguishable from nonspecific interactions that occur between two proteins.

[0056] The term "identical" or percent "identity" in the context of two or more polypeptide sequences refers to two or more sequences or subsequences that are identical or have a certain percentage of identical amino acid residues (e.g., at least 70%, at least 75%, at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more) identity over a particular region, e.g., the length of the two sequences when compared and aligned for maximum correspondence over a comparison window or designated region. Alignment for purposes of determining percent identity of amino acid sequences can be performed in a variety of ways, including, but not limited to, BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. An example of a suitable algorithm for determining percent sequence identity and sequence similarity is the BLAST 2.0 algorithm, which is described in Altschul et al., Nuc. Acids Res. 25:3389-3402 (1977) and Altschul et al., J. Mol. Biol. 215:403-410 (1990). In certain embodiments, BLAST 2.0 can be used with default parameters to determine percent sequence identity.

[0057] As used herein, a "substitution" indicates that one or more amino acids or nucleotides are replaced with different amino acids or nucleotides, respectively.

[0058] As used herein, "conservative" substitution refers to the substitution of amino acids such that charge, polarity, hydrophilicity (hydrophobic, neutral or hydrophilic), and / or size of the side group chain are maintained. Exemplary sets of amino acids that may be substituted for one another include: (i) the positively charged amino acids Lys and Arg; and His at pH about 6; (ii) the negatively charged amino acids Glu and Asp; (iii) the aromatic amino acids Phe, Tyr, Trp; (iv) the nitrogen ring amino acids His and Trp; (v) the aliphatic hydrophobic amino acids Ala, Val, Leu, and Ile; (vi) the hydrophobic sulfur-containing amino acids Met and Cys, which are less hydrophobic than Val, Leu, and Ile; (vii) the small polar uncharged amino acids Ser, Thr, Asp, and Asn; (viii) the small hydrophobic or neutral amino acids Gly, Ala, and Pro; (ix) the amide-containing amino acids Asn and Gln; and (xi) the β-branched amino acids Thr, Val, and Ile. References to the charge of an amino acid refer to the charge at pH 6-7.

[0059] As used herein, the terms "nucleic acid" and "polynucleotide" are used interchangeably and, as used herein, refer to both the sense and antisense strands of RNA, cDNA, genomic DNA, and synthetic forms and mixed polymers of the above. In certain embodiments, polynucleotide refers to polyribonucleotides, polydeoxynucleotides, or modified forms of any type of nucleotide, and combinations thereof. The term also includes, but is not limited to, single-stranded and double-stranded forms of DNA. Furthermore, polynucleotides, such as cDNA or mRNA, may contain either or both naturally occurring and modified nucleotides linked together by naturally occurring and / or non-naturally occurring nucleotide bonds. As will be readily understood by those skilled in the art, nucleic acid molecules may be chemically or biochemically modified or may contain non-natural or derivatized nucleotide bases. Such modifications include, for example, labels, methylation, substitution of one or more analogs of naturally occurring nucleotides, uncharged linkages (e.g., methylphosphonates, phosphotriesters, phosphoramidates, carbamates, etc.), charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.), pendant moieties (e.g., polypeptides), intercalators (e.g., acridines, psoralens, etc.), chelators, alkylators, and internucleotide modifications such as modified linkages (e.g., alpha anomeric nucleic acids, etc.). The above terms are also intended to include any topological conformation, including single-stranded, double-stranded, partially double-stranded, triple-stranded, hairpin, circular, and padlock conformations. Reference to a nucleic acid sequence includes its complement, unless otherwise specified. Thus, reference to a nucleic acid molecule having a particular sequence should be understood to include its complementary strand with its complementary sequence. The term also includes codon-optimized nucleic acids that encode the same polypeptide sequence.

[0060] An "isolated" nucleic acid refers to a nucleic acid molecule that has been separated from a component of its natural environment. Isolated nucleic acid includes a nucleic acid molecule contained within a cell that ordinarily contains the nucleic acid molecule, but where the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0061] An "isolated nucleic acid encoding an antibody or fragment thereof" refers to one or more nucleic acid molecules encoding the heavy and light chains (or fragments thereof) of an antibody, wherein such nucleic acid molecules are contained in a single vector or in separate vectors and are present in one or more locations within a host cell.

[0062] The term "vector" as used herein refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes vectors as self-replicating nucleic acid structures and vectors integrated into the genome of a host cell into which it is introduced. As used herein, "vector" refers to a recombinant construct in which a nucleic acid sequence of interest is inserted into the vector. Certain vectors can direct the expression of a nucleic acid to which it is operably linked. Such vectors are referred to herein as "expression vectors."

[0063] The terms "host cell," "host cell line," and "host cell culture" are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. A host cell can be a recombinant host cell, and includes the primary transformed cell and progeny cells derived therefrom, regardless of the number of passages.

[0064] As used herein, a "variant" of a polypeptide is a polypeptide that typically differs from a polypeptide specifically disclosed herein in one or more substitutions, deletions, additions, and / or insertions. In the present invention, "variants" with respect to the sequences described in the "Anti-CSP antibody variants" section refer to altered sequences rather than naturally occurring sequences.

[0065] In the context of describing the binding strength of two antibodies to the same target, the term "equivalent" refers to the two dissociation constant (KD) values ​​calculated from the two binding reactions being within 3-fold of each other. In certain embodiments, the ratio between the first KD (KD of the binding reaction between the first antibody and the target) and the second KD (KD of the binding reaction between the second antibody and the target) is within the range of 1:3 or 3:1, not including the end points. A lower KD value indicates stronger binding. For example, but not limited to, an antibody variant with stronger binding power compared to AB-000224 binds to the target with a KD that is at least 1 / 3 of the KD of AB-000224 measured against the same target.

[0066] Anti-CSP antibody The present disclosure provides anti-CSP antibodies AB-000224 and AB-007088, and variants thereof. AB-000224 and AB-007088 were discovered from antibody repertoires generated by Immune Repertoire Capture® (IRC®) technology from plasmablast B cells isolated from two donors enrolled in a Phase 2a study evaluating the efficacy of the RTS,S vaccine in preventing malaria infection. IRC® technology and its use in antibody discovery are well known and are disclosed, for example, in WO2012148497A2, the contents of which are incorporated herein by reference in their entirety. The RTS,S vaccine is a pseudovirion vaccine that combines the central repeat and C-terminal regions of the hepatitis B surface antigen and the P. falciparum circumsporozoite protein (CSP). RTS,S consists of two polypeptides; RTS is a single polypeptide chain corresponding to amino acids 207 to 395 of P. falciparum (3D7) fused to HBsAg, and S is a 226 amino acid polypeptide corresponding to HBsAg. Stoute,et.al.,N Engl J Med;336:86-91(1997);RTS,S Clinical Trials Partnership,PLoS Med.11(7:e1001685,(2014),WO1993 / 10152.

[0067] CSP consists of an N-terminal domain containing a heparan sulfate-binding site for hepatocyte adhesion, a central repetitive region, and a structured C-terminal α-thrombospondin repeat (αTSR) followed by a GPI anchor that allows CSP to attach to the sporozoite membrane. The central repetitive region of CSP is highly immunogenic, and in all P. falciparum strains for which CSP sequences are available, the repetitive region is composed of one NPDP repeat, three to five NVDP repeats, and 35 to 41 NANP repeats (e.g., P. falciparum 3D7 strain has a total of 1 / 4 / 38 NPDP / NVDP / NANP motifs). The repetitive region begins with a junctional NPDP sequence, typically followed by three alternating NANP and NVDP sequences, followed by the remaining NANP repeats, although most P. falciparum strains have a single NVDP in the middle of the long NANP repeat region. Pholcharee, T. et al., J.Mol.Bio.432:1048-1063(2020).

[0068] In certain embodiments, the anti-CSP antibodies disclosed herein bind to the central repeat region of P. falciparum CSP. In certain embodiments, the antibodies disclosed herein bind to the P. falciparum CSP protein at the repeat and / or junction region that contains the NPNA, NPDP, and / or NVDP motifs. In certain embodiments, the anti-CSP antibodies disclosed herein bind to the NANP repeat region of P. falciparum CSP. In certain embodiments, the anti-CSP antibodies disclosed herein bind to a polypeptide comprising the amino acid sequence of (NPNA)3 (SEQ ID NO:280).

[0069] In certain embodiments, the disclosure provides anti-CSP antibody variants of AB-000224. In certain embodiments, the disclosure provides anti-CSP antibody variants of AB-007088. In certain embodiments, the variants exhibit a protective effect in vivo, for example, as shown by reduced parasite load in a mouse model of malaria infection.

[0070] In certain embodiments, the anti-CSP variants disclosed herein maintain the binding specificity, activity and stability and / or manufacturing properties of the parent antibody. In certain embodiments, the generated anti-CSP variants disclosed herein have improved exploitability, as identified by various in vitro assays, such as, for example, aggregation assessment by HPLC or UPLC, hydrophobic interaction chromatography (HIC), multispecific assays (e.g., baculovirus particle binding), self-interaction nanoparticle spectroscopy (SINS), or mass spectrometry after incubation under accelerated degradation conditions such as high temperature, low pH, high pH, ​​or oxidizing H2O2. If activity is maintained (or enhanced), while the severity of the disadvantage is eliminated or reduced, the mutation is successful.

[0071] The disadvantages of antibodies are further described in Table 1 below:

[0072] [Table 1]

[0073] 1 "Free cysteine" refers to a cysteine ​​that does not form a disulfide bond with another cysteine ​​and remains "free" as a thiol. The presence of free cysteine ​​in an antibody can be a potential development disadvantage. Typically, an odd total number of cysteines in a protein indicates the possible presence of free cysteine. 2 The N-linked glycosylation sites are NXS / T, where X is any residue other than proline. 3 Sharma et al.,Proc.Natl.Acad.Sci.USA 111:18601-18606,2014. 4 This motif consists of a K or an R followed by a K or an R. In other words, this motif can be KK, KR, RK or RR. 5The dipeptide NG presents a moderate risk of adverse developmental outcomes. The dipeptides NA, NN, NS and NT present a low risk of adverse developmental outcomes. N may also present a low risk of adverse outcomes for other subsequent residues such as D, H or P. Stated differently, the dipeptides ND, NH or NP present a low risk of adverse developmental outcomes. 6 As above, the dipeptide DG poses a moderate risk of adverse developmental outcomes. The dipeptides DA, DD, DS and DT pose a low risk of adverse developmental outcomes. D may also pose a low risk of adverse developmental outcomes for other subsequent residues, e.g., N, H or P.

[0074] Another goal of engineering variants is to reduce the risk of clinical immunogenicity, for example to reduce the generation of anti-drug antibodies against the therapeutic antibody. In certain embodiments, the anti-CSP antibody variants have reduced immunogenicity compared to the parent antibody.

[0075] Factors that promote clinical immunogenicity can be divided into two groups: the first is intrinsic to the drug, such as sequence, post-translational modifications, aggregates, degradation products, and contaminants; the second is related to how the drug is used, such as dosage, frequency of administration, route of administration, immune status of the patient, and HLA type of the patient.

[0076] One approach to making a variant as similar to self as possible is to identify sequences close to the germline and mutate as many mismatched positions as possible (also known as "germline deviations") to the germline residue type. This approach applies to the germline genes IGHV, IGHJ, IGKV, IGKJ, IGLV, and IGLJ, which describe all of the variable heavy region (V11) and variable light region (VL) except for part of the H-CDR3. The germline gene IGHD encodes part of the H-CDR3 region, but typically shows too much variation in how it recombines with IGHV and IGHJ (e.g., forward or reverse, in any of the three translation frames, and in 5' and 3' modifications and untemplated additions) to present a "self" sequence template from a population perspective.

[0077] Each germline gene can appear as different alleles in a population. With a view to minimizing the proportion of patients who have an immunogenic reaction, the least immunogenic drug candidates are likely to match the alleles commonly found in the patient population. Single nucleotide polymorphism (SNP) data from the human genome can be used to estimate the frequency of alleles in a population.

[0078] Another approach to designing leads to reduce the immunogenicity risk is to use in silico prediction of immunogenicity, such as prediction of T cell epitopes, or in vitro assays of immunogenicity, such as ex vivo human T cell activation. Services are available that employ platforms for prediction and in vitro evaluation of HLA binding, such as that offered by Lonza in the UK, to further identify potential epitopes.

[0079] In certain embodiments, antibody variants are further designed to enhance the efficacy of the antibody. The design parameters in this regard focus on CDR, for example CDR3. The positions to be mutated are identified based on the structure analysis of antibody-antigen cocrystals (Oyen et al., Proc. Natl. Acad Sci. USA 114: E10438-E10445, 2017) and based on the sequence information of other antibodies of the same family as AB-000224 or AB-007088.

[0080] 1. Approach to Mutation Design The development disadvantages can be eliminated or mitigated by one or more mutations. The mutations are designed to eliminate or mitigate the development disadvantages and improve function while maintaining the structure and function of the antibody. In certain embodiments, mutations to chemically similar residues are identified to maintain size, shape, charge, and / or polarity. Non-limiting examples of mutations are shown in Table 2 below:

[0081] [Table 2]

[0082] 2. Anti-CSP antibody variants of AB-000224 In certain embodiments, variants of the anti-CSP antibody AB-000224 disclosed herein include modifications relative to AB-000224 that provide improved pharmacokinetic properties, increased serum stability, stronger binding, and / or improved in vivo protective effect compared to AB-000224. In certain embodiments, variants of the anti-CSP antibody AB-000224 disclosed herein exhibit reduced immunogenicity and / or increased manufacturability compared to AB-000224. In certain embodiments, variants of the anti-CSP antibody AB-000224 disclosed herein have at least one modification, e.g., substitution, relative to the native AB-000224 variable heavy chain amino acid sequence (SEQ ID NO: 14) or variable light chain amino acid sequence (SEQ ID NO: 13) described herein, and have improved developability, e.g., reduced heterogeneity, increased yield, increased stability, improved net charge for improved pharmacokinetics, and / or reduced immunogenicity. In certain embodiments, the VH or VL region of such variants of the anti-CSP antibody AB-000224 disclosed herein have at least two, three, four, five, six or more modifications, eg, substitutions. In certain embodiments, variants of anti-CSP antibody AB-000224 disclosed herein have a total of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 modifications, e.g., substitutions, including both variable regions, compared to AB-000224.

[0083] In certain embodiments, variants of the anti-CSP antibody AB-000224 disclosed herein exhibit increased serum half-life compared to AB-000224. In certain embodiments, variants of the anti-CSP antibody AB-000224 disclosed herein have at least one modification, e.g., substitution, to the Fc region of the native AB-000224 heavy chain sequence described herein and have improved pharmacokinetic properties, e.g., half-life. In certain embodiments, the Fc region of the heavy chain of such variants of the anti-CSP antibody AB-000224 disclosed herein has at least two, three, four, five, or six or more modifications, e.g., substitutions. In certain embodiments, variants of anti-CSP antibody AB-000224 disclosed herein have a total of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 modifications, e.g., substitutions, including both heavy and light chains, compared to AB-000224. In certain non-limiting embodiments, the heavy chain Fc region of a variant of the 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, where the numbering is defined by the EU index as in Kabat. In certain embodiments, the heavy chain Fc region of a variant of the anti-CSP antibody AB-000224 disclosed herein can include a leucine at position 428 and a serine at position 434, where the numbering is defined by the EU index as in Kabat.

[0084] The light and heavy chain CDRs of AB-000224 are shown below in Table 3, defined using the Kabat numbering system:

[0085] [Table 3]

[0086] The light and heavy chain CDRs of AB-000224 are defined in the ASN numbering system as shown in Table 4 below:

[0087] [Table 4]

[0088] The sequences of the heavy chain variable region (VH) and light chain variable region (VL) of AB-000224, as well as the sequences of the heavy and light chains, are shown in Table 5 below:

[0089] [Table 5] JPEG2024524895000007.jpg155153

[0090] In certain embodiments, the anti-CSP antibody variant comprises one, two, or three CDRs of the VL sequence of Table 5. In certain embodiments, the anti-CSP antibody variant comprises 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: 13. In certain embodiments, the VL of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, e.g., a substitution, in the CDR1 amino acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the VL of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, e.g., a substitution, in the CDR2 amino acid sequence set forth in SEQ ID NO: 2. In certain embodiments, the VL of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, e.g., a substitution, in the CDR3 amino acid sequence set forth in SEQ ID NO: 3. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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:7. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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:8. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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:9. In certain embodiments, the mutations are conservative substitutions. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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. In certain embodiments, the VL of the anti-CSP antibody variant comprises a substitution at position 1 of SEQ ID NO:13. In certain embodiments, the substitution is E1Q. In certain embodiments, the VL of the anti-CSP antibody variant comprises a substitution at position 4 of SEQ ID NO: 13. In certain embodiments, the substitution is R44T.

[0091] In certain embodiments, the anti-CSP antibody variant comprises one, two, or three CDRs of the VH sequence of Table 5. In certain embodiments, the anti-CSP antibody variant comprises at least one mutation, e.g., a substitution, and no more than 10, 20, 30, 40, or 50 mutations in the VH amino acid sequence set forth in SEQ ID NO: 14. In certain embodiments, the VH of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, e.g., a substitution, in the CDR1 amino acid sequence set forth in SEQ ID NO: 4. In certain embodiments, the VH of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, e.g., a substitution, in the CDR2 amino acid sequence set forth in SEQ ID NO: 5. In certain embodiments, the VH of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, e.g., a substitution, in the CDR3 amino acid sequence set forth in SEQ ID NO: 6. In certain embodiments, the VH of the anti-CSP antibody variant comprises 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: 10. In certain embodiments, the VH of the anti-CSP antibody variant comprises 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: 11. In certain embodiments, the VH of the anti-CSP antibody variant comprises 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: 12. In certain embodiments, the mutations are conservative substitutions. In certain embodiments, the VH of the anti-CSP antibody variant comprises a CDR2 with a substitution at position 12 of SEQ ID NO: 5. In certain embodiments, the substitution is H12K. In certain embodiments, the VH of the anti-CSP antibody variant comprises a CDR2 with a substitution at position 18 of SEQ ID NO: 5. In certain embodiments, the substitution is R18K. In certain embodiments, the mutations are conservative substitutions. In certain embodiments, the VH of the anti-CSP antibody variant comprises a CDR2 with a substitution at position 12 of SEQ ID NO: 11. In certain embodiments, the substitution is H12K.In certain embodiments, the VH of the anti-CSP antibody variant comprises a CDR2 with a substitution at position 18 of SEQ ID NO: 11. In certain embodiments, the substitution is R18K. In certain embodiments, the VH of the anti-CSP antibody variant comprises at least one, at least two, at least three, or at least four mutations, e.g., substitutions, in the framework of the amino acid sequence set forth in SEQ ID NO: 14. An exemplary nucleic acid sequence of SEQ ID NO: 14 is set forth in SEQ ID NO: 16. In certain embodiments, the VH of the anti-CSP antibody variant comprises a substitution at position 21 of SEQ ID NO: 14. In certain embodiments, the substitution is P21S. In certain embodiments, the VH of the anti-CSP antibody variant comprises a substitution at position 23 of SEQ ID NO: 14. In certain embodiments, the substitution is T23A. In certain embodiments, the VH of the anti-CSP antibody variant comprises a substitution at position 80 of SEQ ID NO: 14. In certain embodiments, the substitution is I80T. In certain embodiments, the VH of the variant of the anti-CSP antibody comprises a substitution at position 90 of SEQ ID NO: 14. In certain embodiments, the substitution is T90A. In certain embodiments, the VH of the variant of the anti-CSP antibody comprises a substitution at position 99 of SEQ ID NO: 14. In certain embodiments, the substitution is T99A.

[0092] In certain embodiments, the anti-CSP antibody variant comprises at least one mutation, e.g., a substitution, in the Fc region of the heavy chain amino acid sequence set forth in SEQ ID NO: 18, and comprises no more than 10, 20, 30, 40, or 50 mutations. In certain embodiments, the heavy chain of the anti-CSP antibody variant has the amino acid sequence set forth in SEQ ID NO: 18. An exemplary nucleic acid sequence for SEQ ID NO: 18 is set forth in SEQ ID NO: 21. In certain embodiments, the heavy chain of the anti-CSP antibody variant comprises a substitution at position 438 of SEQ ID NO: 18. In certain embodiments, the substitution is M438L. In certain embodiments, the heavy chain of the anti-CSP antibody variant comprises a substitution at position 444 of SEQ ID NO: 18. In certain embodiments, the substitution is N444S. In certain embodiments, the heavy chain of the anti-CSP antibody variant comprises a substitution at position 438 of SEQ ID NO: 18 and a substitution at position 444 of SEQ ID NO: 18. In certain embodiments, the substitutions are M438L and N444S. In certain embodiments, the heavy chain of an anti-CSP antibody variant has the amino acid sequence set forth in SEQ ID NO: 19. An exemplary nucleic acid sequence for SEQ ID NO: 19 is set forth in SEQ ID NO:22.

[0093] In certain embodiments, the light chain of the anti-CSP antibody AB-000224 and variants thereof comprises a signal peptide. In certain embodiments, the signal peptide is an IGLV2-8 signal peptide. In certain embodiments, the signal peptide has the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the heavy chain of the anti-CSP antibody AB-000224 and variants thereof comprises a signal peptide. In certain embodiments, the signal peptide is an IGKV1-39 signal peptide. In certain embodiments, the signal peptide has the amino acid sequence set forth in SEQ ID NO: 278. SEQ ID NO: 277 and SEQ ID NO: 278 are shown below:

[0094] MAWALLLLTLLTQGTGSWA [SEQ ID NO:277] MDMRVPAQLLGLLLLWLRGARC [SEQ ID NO: 278]

[0095] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 6, and having the amino acid sequence set forth in SEQ ID NO: 23. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 6, and having the amino acid sequence set forth in SEQ ID NO: 24. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 27, as set forth in Table 6. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 28, as set forth in Table 6. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 29, as set forth in Table 6. Exemplary nucleic acid sequences for SEQ ID NOs: 23, 24, 27, 28, and 29 are set forth in Table 6 below.

[0096] [Table 6] JPEG2024524895000009.jpg133153

[0097] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 33, as set forth in Table 7. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 34, as set forth in Table 75. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO: 37, as set forth in Table 7. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 38, as set forth in Table 7. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 39, as set forth in Table 7. Exemplary nucleic acid sequences for SEQ ID NOs: 33, 34, 37, 38, and 39 are set forth in Table 7 below.

[0098] [Table 7] JPEG2024524895000011.jpg169153

[0099] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 8, and having the amino acid sequence set forth in SEQ ID NO: 43. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 8, and having the amino acid sequence set forth in SEQ ID NO: 44. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 47, as set forth in Table 8. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 48, as set forth in Table 8. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 49, as set forth in Table 8. Exemplary nucleic acid sequences for SEQ ID NOs: 43, 44, 47, 48, and 49 are set forth in Table 8 below.

[0100] [Table 8] JPEG2024524895000013.jpg169153

[0101] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 9, and having the amino acid sequence set forth in SEQ ID NO: 53. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 9, and having the amino acid sequence set forth in SEQ ID NO: 55. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 57, as set forth in Table 9. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 58, as set forth in Table 9. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 59, as set forth in Table 9. Exemplary nucleic acid sequences for SEQ ID NOs: 53, 54, 57, 58, and 59 are set forth in Table 9 below.

[0102] [Table 9] JPEG2024524895000015.jpg211153

[0103] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 10, and having the amino acid sequence set forth in SEQ ID NO: 63. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 10, and having the amino acid sequence set forth in SEQ ID NO: 64. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 67, as set forth in Table 10. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 68, as set forth in Table 10. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 69, as set forth in Table 10. Exemplary nucleic acid sequences for SEQ ID NOs: 63, 64, 67, 68, and 69 are set forth in Table 10 below.

[0104] [Table 10] JPEG2024524895000017.jpg221153JPEG2024524895000018.jpg22153

[0105] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 73, as set forth in Table 11. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 74, as set forth in Table 11. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO: 77, as set forth in Table 11. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 78, as set forth in Table 11. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 79, as set forth in Table 11. Exemplary nucleic acid sequences for SEQ ID NOs: 73, 74, 77, 78, and 79 are set forth in Table 11 below.

[0106] [Table 11] JPEG2024524895000020.jpg221153JPEG2024524895000021.jpg29153

[0107] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 12, and having the amino acid sequence set forth in SEQ ID NO: 83. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 12, and having the amino acid sequence set forth in SEQ ID NO: 84. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 87, as set forth in Table 12. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 88, as set forth in Table 12. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 89, as set forth in Table 12. Exemplary nucleic acid sequences for SEQ ID NOs: 83, 84, 87, 88, and 89 are set forth in Table 12 below.

[0108] [Table 12] JPEG2024524895000023.jpg221153JPEG2024524895000024.jpg26153

[0109] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 13, and having the amino acid sequence set forth in SEQ ID NO: 93. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 13, and having the amino acid sequence set forth in SEQ ID NO: 94. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 97, as set forth in Table 13. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 98, as set forth in Table 13. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 99, as set forth in Table 13. Exemplary nucleic acid sequences for SEQ ID NOs: 93, 94, 97, 98, and 99 are set forth in Table 13 below.

[0110] [Table 13] JPEG2024524895000026.jpg221153JPEG2024524895000027.jpg26153

[0111] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 14, and having the amino acid sequence set forth in SEQ ID NO: 103. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 14, and having the amino acid sequence set forth in SEQ ID NO: 104. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 107, as set forth in Table 14. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 108, as set forth in Table 14. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 109, as set forth in Table 14. Exemplary nucleic acid sequences for SEQ ID NOs: 103, 104, 107, 108, and 109 are set forth in Table 14 below.

[0112] [Table 14] JPEG2024524895000029.jpg221153JPEG2024524895000030.jpg26153

[0113] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 113, as set forth in Table 15. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 114, as set forth in Table 15. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO: 117, as set forth in Table 15. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 118, as set forth in Table 15. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 119, as set forth in Table 15. Exemplary nucleic acid sequences for SEQ ID NOs: 113, 114, 117, 118, and 119 are set forth in Table 15 below.

[0114] [Table 15] JPEG2024524895000032.jpg221153JPEG2024524895000033.jpg22153

[0115] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 16, and having the amino acid sequence set forth in SEQ ID NO: 123. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 16, and having the amino acid sequence set forth in SEQ ID NO: 124. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 127, as set forth in Table 16. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 128, as set forth in Table 16. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 129, as set forth in Table 16. Exemplary nucleic acid sequences for SEQ ID NOs: 123, 124, 127, 128, and 129 are set forth in Table 16 below.

[0116] [Table 16] JPEG2024524895000035.jpg221153JPEG2024524895000036.jpg29153

[0117] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 133, as set forth in Table 17. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 134, as set forth in Table 17. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO: 137, as set forth in Table 17. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 138, as set forth in Table 17. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 139, as set forth in Table 17. Exemplary nucleic acid sequences for SEQ ID NOs: 133, 134, 137, 138, and 139 are set forth in Table 17 below.

[0118] [Table 17] JPEG2024524895000038.jpg221153JPEG2024524895000039.jpg29153

[0119] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 143, as set forth in Table 18. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 144, as set forth in Table 18. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO: 147, as set forth in Table 18. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 148, as set forth in Table 18. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 149, as set forth in Table 18. Exemplary nucleic acid sequences for SEQ ID NOs: 143, 144, 147, 148, and 149 are set forth in Table 18 below.

[0120] [Table 18] JPEG2024524895000041.jpg221153JPEG2024524895000042.jpg26153

[0121] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 19, and having the amino acid sequence set forth in SEQ ID NO: 153. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 19, and having the amino acid sequence set forth in SEQ ID NO: 154. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 157, as set forth in Table 19. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 158, as set forth in Table 19. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 159, as set forth in Table 19. Exemplary nucleic acid sequences for SEQ ID NOs: 153, 154, 157, 158, and 159 are set forth in Table 19 below.

[0122] [Table 19] JPEG2024524895000044.jpg221153JPEG2024524895000045.jpg22153

[0123] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 20, and having the amino acid sequence set forth in SEQ ID NO: 163. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 20, and having the amino acid sequence set forth in SEQ ID NO: 164. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 167, as set forth in Table 20. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 168, as set forth in Table 20. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 169, as set forth in Table 20. Exemplary nucleic acid sequences for SEQ ID NOs: 163, 164, 167, 168, and 169 are set forth in Table 20 below.

[0124] [Table 20] JPEG2024524895000047.jpg221153JPEG2024524895000048.jpg22153

[0125] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 173, as set forth in Table 21. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 174, as set forth in Table 21. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 177, as set forth in Table 21. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 178, as set forth in Table 21. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 179, as set forth in Table 21. Exemplary nucleic acid sequences for SEQ ID NOs: 173, 174, 177, 178, and 179 are set forth in Table 21 below.

[0126] [Table 21] JPEG2024524895000050.jpg221153JPEG2024524895000051.jpg22153

[0127] In certain embodiments, the VL of the anti-CSP antibody variant comprises 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: 183. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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: 184. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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: 185. In certain embodiments, the VH of the anti-CSP antibody variant comprises 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: 186. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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: 187. In certain embodiments, the VL of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, eg, substitutions, in the CDR3 amino acid sequence set forth in SEQ ID NO:188.

[0128] In certain embodiments, the VL of the anti-CSP antibody variant comprises 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: 189. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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: 190. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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: 191. In certain embodiments, the VH of the anti-CSP antibody variant comprises 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: 192. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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: 193. In certain embodiments, the VL of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, eg, substitutions, in the CDR3 amino acid sequence set forth in SEQ ID NO:194.

[0129] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 22, and having the amino acid sequence set forth in SEQ ID NO: 195. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 22, and having the amino acid sequence set forth in SEQ ID NO: 196. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 199, as set forth in Table 22. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 200, as set forth in Table 22. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 201, as set forth in Table 22. Exemplary nucleic acid sequences for SEQ ID NOs: 195, 196, 199, 200, and 201 are set forth in Table 22 below.

[0130] [Table 22] JPEG2024524895000053.jpg221153JPEG2024524895000054.jpg22153

[0131] 3. Anti-CSP antibody variants of AB-007088 In certain embodiments, variants of the anti-CSP antibody AB-007088 disclosed herein include modifications relative to AB-007088 that provide improved pharmacokinetic properties, increased serum stability, stronger binding, and / or improved in vivo protective effect compared to AB-007088. In certain embodiments, variants of the anti-CSP antibody AB-007088 disclosed herein exhibit reduced immunogenicity and / or increased manufacturability compared to AB-007088. In certain embodiments, variants of the anti-CSP antibody AB-007088 disclosed herein have at least one modification, e.g., substitution, 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 have improved developability, e.g., reduced heterogeneity, increased yield, increased stability, improved net charge for improved pharmacokinetics, and / or reduced immunogenicity. In certain embodiments, the VH or VL region of such a variant of the anti-CSP antibody AB-007088 disclosed herein has at least two, three, four, five, six or more modifications, eg, substitutions. In certain embodiments, a variant of the 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, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 modifications, e.g., substitutions, including both variable regions, compared to AB-007088.

[0132] In certain embodiments, variants of the anti-CSP antibody AB-007088 disclosed herein exhibit increased serum half-life compared to AB-007088. In certain embodiments, variants of the anti-CSP antibody AB-007088 disclosed herein have at least one modification, e.g., substitution, to the Fc region of the native AB-007088 heavy chain herein and have improved pharmacokinetic properties, e.g., half-life. In certain embodiments, the Fc region of the heavy chain of such variants of the anti-CSP antibody AB-007088 disclosed herein has at least two, three, four, five, or six or more modifications, e.g., substitutions. In certain embodiments, variants of anti-CSP antibody AB-007088 disclosed herein have a total of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 modifications, e.g., substitutions, including both heavy and light chains, compared to AB-007088. In certain non-limiting embodiments, the heavy chain Fc region of a variant of the anti-CSP antibody AB-007088 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, where the numbering is defined by the EU index as in Kabat. In certain embodiments, the heavy chain Fc region of a variant of the anti-CSP antibody AB-007088 disclosed herein can include a leucine at position 428 and a serine at position 434, where the numbering is defined by the EU index as in Kabat.

[0133] The light and heavy chain CDRs of AB-007088 are shown below in Table 23, defined using the Kabat numbering system:

[0134] [Table 23]

[0135] The light and heavy chain CDRs of AB-007088, defined by ASN, are shown below in Table 24:

[0136] [Table 24]

[0137] The sequences of the heavy chain variable region (VH) and light chain variable region (VL) of AB-007088, as well as the sequences of the heavy and light chains, are shown in Table 25 below:

[0138] [Table 25] JPEG2024524895000058.jpg195153

[0139] In certain embodiments, the anti-CSP antibody variant comprises one, two, or three CDRs of the VL sequence of Table 25. In certain embodiments, the anti-CSP antibody variant comprises 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. In certain embodiments, the VL of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, e.g., a substitution, in the CDR1 amino acid sequence set forth in SEQ ID NO:205. In certain embodiments, the VL of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, e.g., a substitution, in the CDR2 amino acid sequence set forth in SEQ ID NO:206. In certain embodiments, the VL of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, e.g., a substitution, in the CDR3 amino acid sequence set forth in SEQ ID NO:207. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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: 211. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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: 212. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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: 213. In certain embodiments, the mutations are conservative substitutions. In certain embodiments, the VL of the anti-CSP antibody variant comprises 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.

[0140] In certain embodiments, the anti-CSP antibody variant comprises one, two, or three CDRs of the VH sequence of Table 25. In certain embodiments, the anti-CSP antibody variant comprises at least one mutation, e.g., a substitution, and no more than 10, 20, 30, 40, or 50 mutations in the VH amino acid sequence set forth in SEQ ID NO:218. In certain embodiments, the VH of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, e.g., a substitution, in the CDR1 amino acid sequence set forth in SEQ ID NO:208. In certain embodiments, the VH of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, e.g., a substitution, in the CDR2 amino acid sequence set forth in SEQ ID NO:209. In certain embodiments, the VH of the anti-CSP antibody variant comprises at least one, at least two, or at least three mutations, e.g., a substitution, in the CDR3 amino acid sequence set forth in SEQ ID NO:210. In certain embodiments, the VH of the anti-CSP antibody variant comprises 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 anti-CSP antibody variant comprises 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. In certain embodiments, the VH of the anti-CSP antibody variant comprises 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. In certain embodiments, the mutations are conservative substitutions. In certain embodiments, the VH of the anti-CSP antibody variant comprises 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. In certain embodiments, the VH of the anti-CSP antibody variant comprises a substitution at position 28 of SEQ ID NO: 218. In certain embodiments, the substitution is A28T. In certain embodiments, the VH of the anti-CSP antibody variant comprises a substitution at position 40 of SEQ ID NO: 218.In certain embodiments, the substitution is T40A. In certain embodiments, the VH of the anti-CSP antibody variant comprises a substitution at position 69 of SEQ ID NO:218. In certain embodiments, the substitution is I69T. In certain embodiments, the VH of the anti-CSP antibody variant comprises a substitution at position 80 of SEQ ID NO:218. In certain embodiments, the substitution is S80Y. In certain embodiments, the VH of the anti-CSP antibody variant comprises a substitution at position 85 of SEQ ID NO:218. In certain embodiments, the substitution is G85S. In certain embodiments, the VH of the anti-CSP antibody variant comprises a substitution at position 120 of SEQ ID NO:218. In certain embodiments, the substitution is I120T.

[0141] In certain embodiments, the light chain of the variant of the 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. In certain embodiments, the variant of the anti-CSP antibody comprises at least one mutation, e.g., substitution, in the Fc region of the heavy chain amino acid sequence set forth in SEQ ID NO: 222, and comprises no more than 10, 20, 30, 40, or 50 mutations. In certain embodiments, the heavy chain of the variant of the 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. In certain embodiments, the heavy chain of the variant of the anti-CSP antibody comprises a substitution at position 434 of SEQ ID NO: 222. In certain embodiments, the substitution is M434L. In certain embodiments, the heavy chain of the variant of the anti-CSP antibody comprises a substitution at position 440 of SEQ ID NO: 222. In certain embodiments, the substitution is L440S. In certain embodiments, the heavy chain of the variant of the anti-CSP antibody comprises a substitution at position 434 of SEQ ID NO:222 and a substitution at position 440 of SEQ ID NO:222. In certain embodiments, the substitutions are M434L and N440S. In certain embodiments, the heavy chain of the variant of the anti-CSP antibody has the amino acid sequence set forth in SEQ ID NO:223. An exemplary nucleic acid sequence of SEQ ID NO:223 is set forth in SEQ ID NO:226.

[0142] In certain embodiments, the light chain of the anti-CSP antibody AB-007088 and variants thereof comprises a signal peptide. In certain embodiments, the signal peptide is an IGLV2-8 signal peptide. In certain embodiments, the signal peptide has the amino acid sequence set forth in SEQ ID NO: 277. In certain embodiments, the heavy chain of the anti-CSP antibody AB-007088 and variants thereof comprises a signal peptide. In certain embodiments, the signal peptide is an IGKV1-39 signal peptide. In certain embodiments, the signal peptide has the amino acid sequence set forth in SEQ ID NO: 278.

[0143] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 26, and having the amino acid sequence set forth in SEQ ID NO: 227. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 26, and having the amino acid sequence set forth in SEQ ID NO: 228. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 231, as set forth in Table 26. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 232, as set forth in Table 26. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 233, as set forth in Table 26. Exemplary nucleic acid sequences for SEQ ID NOs: 227, 228, 231, 232, and 233 are set forth in Table 26 below.

[0144] [Table 26] JPEG2024524895000060.jpg221153JPEG2024524895000061.jpg22153

[0145] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 27, and having the amino acid sequence set forth in SEQ ID NO: 237. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 27, and having the amino acid sequence set forth in SEQ ID NO: 238. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 241, as set forth in Table 27. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 242, as set forth in Table 27. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 243, as set forth in Table 27. Exemplary nucleic acid sequences for SEQ ID NOs: 237, 238, 241, 242, and 243 are set forth in Table 27 below.

[0146] [Table 27] JPEG2024524895000063.jpg221153JPEG2024524895000064.jpg22153

[0147] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 247, as set forth in Table 28. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3 and having the amino acid sequence set forth in SEQ ID NO: 248, as set forth in Table 28. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 251, as set forth in Table 28. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 252, as set forth in Table 28. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 253, as set forth in Table 28. Exemplary nucleic acid sequences for SEQ ID NOs: 247, 248, 251, 252, and 253 are set forth in Table 28 below.

[0148] [Table 28] JPEG2024524895000066.jpg221153JPEG2024524895000067.jpg22153

[0149] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 29, and having the amino acid sequence set forth in SEQ ID NO: 257. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 29, and having the amino acid sequence set forth in SEQ ID NO: 258. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 261, as set forth in Table 29. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 262, as set forth in Table 29. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 263, as set forth in Table 29. Exemplary nucleic acid sequences for SEQ ID NOs: 257, 258, 261, 262, and 263 are set forth in Table 29 below.

[0150] [Table 29] JPEG2024524895000069.jpg221153JPEG2024524895000070.jpg22153

[0151] In certain embodiments, the anti-CSP antibody variant comprises a light chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 30, and having the amino acid sequence set forth in SEQ ID NO: 267. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3, as set forth in Table 30, and having the amino acid sequence set forth in SEQ ID NO: 268. In certain embodiments, the anti-CSP antibody variant comprises a light chain comprising an amino acid sequence set forth in SEQ ID NO: 271, as set forth in Table 30. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 272, as set forth in Table 30. In certain embodiments, the anti-CSP antibody variant comprises a heavy chain comprising an amino acid sequence set forth in SEQ ID NO: 273, as set forth in Table 30. Exemplary nucleic acid sequences for SEQ ID NOs: 267, 268, 271, 272, and 273 are set forth in Table 30 below.

[0152] [Table 30] JPEG2024524895000072.jpg221153JPEG2024524895000073.jpg22153

[0153] 4. Glycosylation of Anti-CSP Antibodies and Their Variants Glycosylation of antibodies and modified antibodies has been disclosed previously (see, e.g., U.S. Patent No. 6,602,684, the contents of which are incorporated in their entirety). Antibody Fc regions are generally post-translationally modified by the addition of N-glycans at specific asparagine residues on the antibody heavy chain. IgG molecules have an N-linked glycosylated asparagine on each heavy chain. It has been shown that modified glycosylation profiles can modulate antibody function. For example, but not limited to, altered glycosylation can improve the binding affinity or half-life of an antibody compared to unmodified versions.

[0154] In certain embodiments, the present disclosure provides anti-CSP antibodies and variants thereof with modified glycosylation. In certain embodiments, the antibodies disclosed herein comprise an Fc region with increased glycosylation. In certain non-limiting embodiments, the Fc region with increased glycosylation comprises an increased amount of bisected oligosaccharides. In certain embodiments, the Fc region with increased glycosylation comprises an increased amount of non-fucosylated oligosaccharides. In certain embodiments, the Fc region with increased glycosylation comprises an increased amount of fucose-containing oligosaccharides.

[0155] In certain embodiments, the antibodies disclosed herein comprise an Fc region with reduced glycosylation. In certain non-limiting embodiments, an Fc region with reduced glycosylation comprises a reduced amount of bisected oligosaccharides. In certain embodiments, an Fc region with reduced glycosylation comprises a reduced amount of nonfucosylated oligosaccharides. In certain embodiments, an Fc region with increased glycosylation comprises a reduced amount of fucose-containing oligosaccharides.

[0156] In certain embodiments, the antibodies disclosed herein comprise a V region with increased glycosylation. In certain non-limiting embodiments, the V region with increased glycosylation comprises an increased amount of bisected oligosaccharides. In certain embodiments, the V region with increased glycosylation comprises an increased amount of non-fucosylated oligosaccharides. In certain embodiments, the V region with increased glycosylation comprises an increased amount of fucose-containing oligosaccharides.

[0157] In certain embodiments, the antibodies disclosed herein comprise a V region with reduced glycosylation. In certain non-limiting embodiments, a V region with reduced glycosylation comprises a reduced amount of bisected oligosaccharides. In certain embodiments, a V region with reduced glycosylation comprises a reduced amount of nonfucosylated oligosaccharides. In certain embodiments, a V region with increased glycosylation comprises a reduced amount of fucose-containing oligosaccharides.

[0158] In certain embodiments, modified glycosylation can be obtained by expressing any of the antibodies disclosed herein in a host cell with altered glycosylation machinery. For example, but not limited to, the host cell can contain a functional disruption of a fucosyltransferase gene, and antibodies expressed in the host cell exhibit reduced glycosylation, e.g., reduced fucosylation (see PCT Patent Publication No. WO 99 / 54342).

[0159] In certain embodiments, the disclosure provides an anti-CSP antibody variant disclosed herein that comprises one or more amino acid substitutions that result in an alteration of a glycosylation acceptor site. In certain embodiments, the alteration comprises removal of a glycosylation acceptor site. In certain embodiments, the alteration comprises modification of a glycosylation acceptor site. In certain embodiments, the alteration comprises insertion of a glycosylation acceptor site.

[0160] As used herein, "glycosylation acceptor site" refers to an amino acid residue in the light or heavy chain of an antibody that can be N-glycosylated or O-glycosylated. In certain embodiments, the N-linked glycosylation acceptor site can be an asparagine residue. In certain embodiments, the O-linked glycosylation acceptor site can be a serine, threonine, tyrosine, hydroxylysine, or hydroxyproline residue.

[0161] In certain embodiments, the Fc region of the antibodies disclosed herein comprises one or more glycosylation acceptor sites. In certain embodiments, the V region of any of the antibodies disclosed herein comprises one or more glycosylation acceptor sites. In certain embodiments, the light chain of any of the antibodies disclosed herein comprises one or more glycosylation acceptor sites. In certain embodiments, the heavy chain of any of the antibodies disclosed herein comprises one or more glycosylation acceptor sites. In certain embodiments, the light chain variable region of any of the antibodies disclosed herein comprises one or more glycosylation acceptor sites. In certain embodiments, the heavy chain variable region of any of the antibodies disclosed herein comprises one or more glycosylation acceptor sites.

[0162] 5. PEGylation and Other Chemical Modifications of Anti-CSP Antibodies and Their Variants The present disclosure provides anti-CSP antibodies and variants thereof that contain additional modifications. In certain embodiments, the modifications can improve the pharmacological properties of the antibody, such as half-life. In certain non-limiting embodiments, the modifications include PEGylation, deamination, derivatization with polymers, lipidation, removal and / or introduction of disulfide bonds, oxidation, and removal of C-terminal lysine.

[0163] In certain embodiments, the modification is PEGylation. PEGylation of antibodies and modified antibodies includes conjugating one or more polyethylene glycols (PEGs) to the antibody. In certain non-limiting embodiments, for example, PEGylation can be carried out by an acylation reaction or an alkylation reaction with a reactive PEG molecule (or an analogous reactive water-soluble polymer). As used herein, the term "polyethylene glycol" refers to any of the forms of PEG that have been used to derivatize other proteins, such as mono (C1-C10) alkoxy- or aryloxy-polyethylene glycol or polyethylene glycol-maleimide.

[0164] In certain embodiments, the modification is derivatization with hydrophilic polymer.In certain non-limiting embodiments, for example, the hydrophilic polymer can be carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1,3-dioxolane, poly-1,3,6-trioxane, ethylene / maleic anhydride copolymer, polyamino acid (either homopolymer or random copolymer), and dextran or poly(n-vinylpyrrolidone) polyethylene glycol, propylene glycol homopolymer, propylene oxide / ethylene oxide copolymer, polyoxyethylated polyol (e.g., glycerol), polyvinyl alcohol, and mixtures thereof.

[0165] In certain embodiments, the modification is lipidation. Lipidation is the conjugation of proteins and lipids. Lipidation of peptides improves metabolic stability, membrane permeability, bioavailability, and changes the pharmacokinetic and pharmacodynamic properties of peptides. For example, lipidated peptides have high affinity with serum albumin, resulting in increased half-life and stability. In certain non-limiting embodiments, for example, lipids can be myristic acid, palmitic acid, stearic acid, lauric acid, cholesterol, and mixtures thereof.

[0166] In certain embodiments, the modification is a substitution of an amino acid residue to form a disulfide bond. In certain embodiments, the amino acid substitution introduces a cysteine. Under certain redox conditions, two cysteines can form a non-natural disulfide bond. In certain non-limiting embodiments, the disulfide bond improves the stability of the antibody, e.g., corrects the pairing of the antibody chains. In certain embodiments, the cysteine ​​is introduced in the V region. In certain embodiments, the cysteine ​​is introduced in the Fc region. In certain embodiments, the modification is a substitution of an amino acid residue to remove a disulfide bond. In certain embodiments, the amino acid substitution removes a cysteine. In certain embodiments, the cysteine ​​is replaced with a serine. In certain non-limiting embodiments, removing the cysteine ​​improves the stability of the antibody, e.g., improves the long-term stability. In certain embodiments, the cysteine ​​is removed in the V region. In certain embodiments, the cysteine ​​is removed in the Fc region.

[0167] 6. Conjugates of Anti-CSP Antibodies and Anti-CSP Antibody Variants In certain embodiments, the present disclosure provides an anti-CSP antibody or variant thereof conjugated or linked to a therapeutic moiety and / or an imaging / detectable moiety. For example, but not limited to, the anti-CSP antibody or variant thereof may be conjugated to a detectable marker, a toxin, or a therapeutic agent. The moiety may be covalently or non-covalently linked to the antibody.

[0168] In certain embodiments, the antibody or variant thereof is conjugated to a cytotoxic moiety or other moiety that inhibits cell proliferation. In certain embodiments, the antibody or variant thereof is conjugated to a cytotoxic moiety or other moiety that inhibits cell proliferation, such as ricin A chain, doxorubicin, daunorubicin, maytansinoid taxol, ethidium bromide, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicine, dihydroxyanthracin dione, actinomycin, diphtheria toxin, Pseudomonas exotoxin A, Pseudomonas exotoxin (PE) A, PE40, abrin, abrin A chain, modectin A chain, alphasarcin. , gelonin, mitogenin, restrictocin, cobra venom factor, ribonuclease, phenomycin, enomycin, curicin, crotin, calicheamicin, Saponaria officinalis inhibitor, glucocorticoid, auristatin, auromycin, yttrium, bismuth, combrestatin, duocarmycin, dolastatin, cc1065, or cisplatin. In certain embodiments, the antibody or variant thereof may be linked to an agent such as an enzyme inhibitor, a proliferation inhibitor, a lytic agent, a DNA or RNA synthesis inhibitor, a membrane permeability modifier, a DNA metabolite, a dichloroethylsulfide derivative, a protein production inhibitor, a ribosome inhibitor, or an apoptosis inducer.

[0169] In certain embodiments, the antibody or variants thereof may be linked to a radionuclide, an iron-related compound, a dye, a fluorescent agent, or an imaging agent. In certain embodiments, the antibody may be linked to an agent such as, but not limited to, a metal; a metal chelator; a lanthanide; a lanthanide chelator; a radioactive metal; a radioactive metal chelator; a positron-emitting nucleus; a microbubble (for ultrasound); a liposome; a molecule microencapsulated in a liposome or nanosphere; a single crystal iron oxide nanocompound; a magnetic resonance imaging contrast agent; a light absorbing agent, a light reflecting agent, and / or a scattering agent; a colloidal particle; a fluorophore, such as a near-infrared fluorophore.

[0170] In certain embodiments, the present disclosure provides bispecific molecules comprising the anti-CSP antibodies, variants thereof, or fragments thereof disclosed herein. The anti-CSP antibodies, anti-CSP antibody variants, or antigen-binding portions thereof, can be derivatized or linked to another functional molecule, such as another peptide or protein (e.g., a ligand for another antibody or receptor), to generate bispecific molecules that bind at least two different binding sites or target molecules. The anti-CSP antibodies or variants thereof disclosed herein can be derivatized or linked to one or more other functional molecules to generate multispecific molecules that bind to two or more different binding sites (e.g., two different epitopes on a CSP protein) and / or target molecules; such multispecific molecules are also intended to be encompassed by the term "bispecific molecule" as used herein. To generate the bispecific molecules of the present invention, the antibodies of the present invention can be functionally linked (e.g., by chemical coupling, genetic fusion, non-covalent binding, etc.) to one or more other binding molecules, such as another antibody, antibody fragment, peptide, or binding mimic. In certain non-limiting embodiments, for example, but not limited to, bispecific antibodies can be made using a knobs-in-holes strategy. This strategy typically involves making a first half of a first antibody that recognizes a first antigen, e.g., CSP, and a second half of an antibody that recognizes a second antigen or binding site, and then combining the two halves to make a bispecific antibody. In certain embodiments, the first and second antigens are different epitopes of the CSP protein.

[0171] 7. Activity The activity of the anti-CSP antibodies disclosed herein can be evaluated using different endpoints. In certain embodiments, activity is evaluated for binding to CSP, either a series of linear peptides of various lengths representing the immunodominant region of the CSP protein, or binding to the entire CSP protein. In certain embodiments, activity is evaluated for protection against challenge with Plasmodium containing the CSP of P. falciparum, for example in an in vivo animal model of malaria. In certain embodiments, effector function, such as ADCC, is also evaluated.

[0172] In certain embodiments, the binding activity of the anti-CSP antibodies disclosed herein to P. falciparum CSP protein can be assessed by surface plasmon resonance (SPR) using a biosensor system. Systems suitable for use in SPR include, but are not limited to, LSATM (Carterra, Dublin, CA), Biacore™ (General Electric, Boston, MA), and OpenSPR (Nicoya, East Kitchener, ON, Canada). In an exemplary SPR assay, each antibody can be directly immobilized on a Carterra CMD200M chip or captured on a CMD200M Carterra Chip with a goat anti-human IgG Fc antibody. Uncoupled antibodies can be washed away and various concentration gradients of targets can be flowed over the antibodies. Under certain experimental conditions, the highest concentration of each target can range from 0.5 to 8 μg / mL. For greater precision, each antibody can be immobilized at a different location (e.g., at least two locations) on the chip and the affinity for each antibody-target combination can be determined using multiple (e.g., 4-5) target concentrations according to standard methods. If the variation between the two duplicates exceeds 3-fold, the antibody-target measurements are repeated.

[0173] In certain embodiments, the binding activity of the anti-CSP antibodies disclosed herein to P. falciparum CSP protein can be assessed by biolayer interferometry (BLI). In BLI, each antigen (e.g., those disclosed in Table 35) can be immobilized on a sensor according to the manufacturer's instructions. Systems suitable for use in BLI include, but are not limited to, Octet™ (ForteBio, Fremont, CA) and Gator™ (Probelife, Palo Alto, CA). In certain embodiments, for example, but not limited to, the antigen can be biotinylated and immobilized on a streptavidin sensor. For greater accuracy, each antibody can be assessed in duplicate at an appropriate concentration (e.g., 5 μg / mL). If the variation between the two duplicates is more than three-fold, the antibody-target measurement is repeated. The assay is typically performed under conditions according to the manufacturer's instructions. The assay can be performed at a temperature ranging from 20° C. to 37° C., for example, from 20° C. to 25° C. In certain embodiments, the assay is performed at 25° C. In certain embodiments, the assay is performed at 37° C.

[0174] In certain embodiments, binding to a CSP protein is assessed in a competitive assay format using reference antibody AB-000224 or a reference antibody having the variable region of AB-000224. In certain embodiments, binding to a CSP protein is assessed in a competitive assay format using reference antibody AB-007088 or a reference antibody having the variable region of AB-007088. In certain embodiments, the variant anti-CSP antibodies disclosed herein may block binding of the reference antibody by about 50% or more in a competitive assay.

[0175] The anti-CSP antibodies and anti-CSP antibody variants of the present disclosure may also be evaluated in various assays for their ability to mediate FcR-dependent activities. In certain embodiments, the antibodies of the present disclosure have enhanced ADCC and / or serum stability compared to antibody AB-000224 when the antibodies are assayed in a human IgG1 isotype format. In certain embodiments, the antibodies of the present disclosure have enhanced ADCC and / or serum stability compared to antibody AB-007088 when the antibodies are assayed in a human IgG1 isotype format.

[0176] In certain embodiments, the activity of anti-CSP antibodies can be evaluated in vivo in animal models, for example, as described in the Examples section. In certain non-limiting embodiments, a mouse malaria liver burden assay can be used, for example, as disclosed in Flores-Garcia Y, et al. Malar J. 2019;18(1):426, doi:10.1186 / s12936-019-3055-9, the contents of which are incorporated herein by reference. Mice are administered the antibody and infected with a chimeric P. berghei expressing GFP-luciferase and the CSP protein of P. falciparum. Parasite liver burden can be evaluated, for example, by measuring bioluminescence by RT-qPCR or IVIS Spectrum imager. Reduction in parasite liver burden reflects the prophylactic activity of the antibody. In certain embodiments, the activity of anti-CSP antibodies can be determined by evaluating in vivo protection and survival in animal models, e.g., mice. For example, but not limited to, mice are administered the antibody and challenged with chimeric P. berghei expressing the CSP protein of P. falciparum, as disclosed in, for example, Espinosa, D., et al. npj Vaccines 2017;2,10(2017); Espinosa, D., et al. Infect Immun. 2013 Aug;81(8):2882-2887. In vivo prophylactic efficacy can be determined by detecting blood-stage parasitemia by microscopic examination. Survival can be determined by utilizing the absence of parasitemia during an observation period (e.g., 2 weeks) immediately following challenge. Increased survival reflects the prophylactic and / or therapeutic activity of the antibody.

[0177] In certain embodiments, the anti-CSP antibodies disclosed herein, such as AB-000224 or AB-007088, have at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or 70% or more of the activity of antibody AB-000317 when evaluated under the same assay conditions. In certain embodiments, the anti-CSP antibodies exhibit improved activity, i.e., greater than 100%, compared to antibody AB-000317. In certain non-limiting embodiments, the anti-CSP antibodies disclosed herein exhibit at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or 70% or more reduction in parasite liver burden compared to antibody AB-000317. In certain non-limiting embodiments, the anti-CSP antibodies disclosed herein exhibit an increase in survival rate of at least 20%, or at least 30%, or at least 40%, or at least 50%, or at least 60%, or 70%, or more, compared to antibody AB-000317.

[0178] In certain embodiments, the anti-CSP antibody variants disclosed herein have at least 50%, or at least 60%, or 70% or more of the activity of AB-000224 when evaluated under the same assay conditions. In certain embodiments, the anti-CSP antibody variants disclosed herein have improved activity compared to AB-000224, i.e., greater than 100% activity. In certain embodiments, the anti-CSP antibody variants disclosed herein have similar activity against malaria infection compared to AB-000224. In certain embodiments, the anti-CSP antibody variants disclosed herein have at least 50%, or at least 60%, or 70% or more of the activity of AB-007088 when evaluated under the same assay conditions. In certain embodiments, the anti-CSP antibody variants disclosed herein have improved activity compared to AB-007088, i.e., greater than 100% activity. In certain embodiments, the anti-CSP antibody variants disclosed herein have similar activity against malaria infection compared to AB-007088. When used to compare the in vivo activity of antibodies, the term "similar activity" means that two measurements of activity differ from each other by no more than 30%, no more than 25%, no more than 20%, no more than 15%, no more than 10%, no more than 8%, or no more than 5%.

[0179] In certain embodiments, the native anti-CSP antibody AB-000224 is modified to have improved developability (i.e., reduced developmental disadvantages), including but not limited to reduced heterogeneity, increased yield, increased stability, improved net charge to improve pharmacokinetics, and / or reduced immunogenicity. In certain embodiments, antibodies with improved developability can be obtained by introducing mutations that reduce or eliminate potential developmental disadvantages, as described in Table 1. In certain embodiments, antibodies with improved developability have modifications in the amino acid sequence compared to AB-000224, as disclosed in Table 2.

[0180] In certain embodiments, the native anti-CSP antibody AB-007088 is modified to have improved developability (i.e., reduced developmental disadvantages), including but not limited to reduced heterogeneity, increased yield, increased stability, improved net charge to improve pharmacokinetics, and / or reduced immunogenicity. In certain embodiments, antibodies with improved developability can be obtained by introducing mutations that reduce or eliminate potential developmental disadvantages, as described in Table 1. In certain embodiments, antibodies with improved developability have modifications in the amino acid sequence compared to AB-007088, as disclosed in Table 2.

[0181] In certain embodiments, the anti-CSP antibody variants disclosed herein have improved developability while maintaining the same or improved binding affinity to the target compared to AB-000224. Non-limiting examples of such anti-CSP antibody variants are disclosed herein. In certain embodiments, the anti-CSP antibody variants disclosed herein have improved developability while maintaining similar activity to AB-000224.

[0182] In certain embodiments, the anti-CSP antibody variants disclosed herein have improved developability while maintaining the same or improved binding affinity to the target compared to AB-007088. Non-limiting examples of such anti-CSP antibody variants are disclosed herein. In certain embodiments, the anti-CSP antibody variants disclosed herein have improved developability while maintaining similar activity to AB-007088.

[0183] 8. Antibody Generation The CSP antibodies and variants thereof disclosed herein can be produced using vectors and recombinant techniques (see, e.g., Sambrook & Russell, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press; Ausubel, Current Protocols in Molecular Biology). Reagents, cloning vectors, and kits for genetic manipulation are available from commercial vendors.

[0184] The present disclosure provides isolated nucleic acids encoding the VH and / or VL regions, or fragments thereof, of any of the anti-CSP antibodies and anti-CSP antibody variants disclosed herein. In certain embodiments, the present disclosure provides vectors comprising the nucleic acids used to replicate the nucleic acids encoding the antibodies and / or to express the antibodies, and host cells into which the nucleic acids are introduced. These nucleic acids may encode an amino acid sequence comprising the VL and / or an amino acid sequence comprising the VH of an anti-CSP antibody or variant thereof (e.g., the light and / or heavy chains of the antibody). In certain embodiments, the host cell comprises (1) a vector comprising a polynucleotide encoding a VL amino acid sequence and a polynucleotide encoding a VH amino acid sequence, or (2) a first vector comprising a polynucleotide encoding a VL amino acid sequence and a second vector comprising a polynucleotide encoding a VH amino acid sequence.

[0185] In certain embodiments, the present disclosure provides a method of making an anti-CSP antibody disclosed herein. In certain embodiments, the method comprises culturing the aforementioned host cell under conditions suitable for expression of the antibody. In certain embodiments, the antibody is then recovered from the host cell (or host cell culture medium).

[0186] Suitable vectors containing polynucleotides encoding the antibodies or fragments thereof of the present disclosure include cloning vectors and expression vectors. The cloning vector selected may vary depending on the host cell used, but useful cloning vectors are generally capable of autonomous replication, may have a single target for a specific restriction endonuclease, and / or may carry a marker gene that can be used in selecting clones containing the vector. Non-limiting examples include plasmids and bacterial viruses, such as pUC18, pUC19, Bluescript (e.g., pBS SK+) and its derivatives, mp18, mp19, pBR322, pMB9, ColEl plasmids, pCR1, RP4, phage DNA, and shuttle vectors.

[0187] Expression vector is generally a replicable polynucleotide construct that contains the nucleic acid of the present disclosure.Expression vector can replicate in host cell as episome or as part of chromosomal DNA.Suitable expression vectors include, but are not limited to, plasmid and viral vectors, including adenovirus, adeno-associated virus, retrovirus and other vectors.

[0188] Suitable host cells for expressing the anti-CSP antibodies or anti-CSP antibody variants disclosed herein include both prokaryotic and eukaryotic cells. For example, but not limited to, anti-CSP antibodies can be produced in bacteria, especially when glycosylation and Fc effector functions are not required. After expression, the antibody can be isolated as a soluble fraction from bacterial cell lysates and further purified. Alternatively, the host cell can be a eukaryotic host cell, including but not limited to eukaryotic microorganisms such as filamentous fungi or yeast, fungal or yeast strains in which the glycosylation pathway has been "humanized" to result in the production of antibodies with partially or fully human glycosylation patterns, vertebrate, invertebrate, and plant cells. Non-limiting examples of invertebrate cells include insect cells. Numerous baculovirus strains have been identified that can be used in combination with insect cells. Plant cell cultures can also be utilized as host cells.

[0189] In certain embodiments, vertebrate host cells are used to produce the anti-CSP antibody of the present disclosure.For example, but not limited to, mammalian cell lines that can be used to express anti-CSP antibody include SV40 transformed monkey kidney CV1 line (COS-7); human embryonic kidney line (293 or 293 cells); baby hamster kidney cells (BHK); mouse Sertoli cells (TM4 cells); monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); dog kidney cells (MDCK); buffalo rat hepatocytes (BRL3A); human lung cells (W138); human hepatocytes (HepG2); mouse mammary tumor (MMT060562); TRI cells; MRC5 cells; and FS4 cells. In certain embodiments, the mammalian cell line used to express the anti-CSP antibody may be a Chinese hamster ovary (CHO) cell line; a DHFR-CHO cell line (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216, 1980); and myeloma cell lines such as YO, NSO, and Sp2 / 0. Host cells of the present disclosure also include, but are not limited to, isolated cells, in vitro cultured cells, and ex vivo cultured cells.

[0190] Host cells transfected with an expression vector encoding an anti-CSP antibody or fragment thereof of the present disclosure can be cultured under appropriate conditions to allow expression of the polypeptide to occur. The polypeptide can be secreted and isolated from a mixture of cells and medium containing the polypeptide. Alternatively, the polypeptide can be retained in the cytoplasm or a membrane fraction, and the cells can be harvested, lysed, and the polypeptide isolated in a desired manner.

[0191] In certain embodiments, the present disclosure provides a method for making variants of the anti-CSP antibodies disclosed herein. In certain non-limiting embodiments, constructs encoding variants of VH CDR2, such as, for example, but not limited to, those described in the "Anti-CSP Antibody Variants" section, may be additionally modified, and the VH region encoded by the additionally modified construct may be tested for binding activity and / or in vivo protective efficacy against CSP in the context of a VH region comprising native AB-000224 CDR1 and CDR3, or a variant CDR1 or CDR3 described herein (paired with native AB-000224 VL region or variant region described herein). Similarly, constructs encoding variant VL CDR3, such as those described in the "Anti-CSP Antibody Variants" section, may be additionally modified, and the VL region encoded by the additionally modified construct may be tested for binding activity and / or protective efficacy against CSP. Also, such analyses may be performed on other CDRs or framework regions, and antibodies with the desired activity may be selected.

[0192] Pharmaceutical Compositions and Methods of Treatment In certain embodiments, the present disclosure provides pharmaceutical compositions for administration of anti-CSP antibodies and variants thereof. In certain embodiments, the pharmaceutical compositions may be administered to a mammalian subject, e.g., a human, suffering from or at risk of malaria in a therapeutically effective amount according to a schedule sufficient to prevent Plasmodium infection, e.g., infection with Plasmodium falciparum or Plasmodium sp. having a cross-reactive CSP protein, or sufficient to reduce symptoms of malaria in the subject. In certain embodiments, the pharmaceutical composition may include any of the anti-CSP antibodies and variants thereof disclosed herein, or a polynucleotide encoding same, and a pharmaceutically acceptable diluent or carrier. In certain embodiments, the polynucleotide encoding the antibody may be included in a plasmid vector or a viral vector for delivery. In certain embodiments, the pharmaceutical composition includes a therapeutically effective amount of the antibody. As used herein, a "therapeutically effective dose" or a "therapeutically effective amount" refers to an amount sufficient to prevent, cure, or at least partially arrest malaria or symptoms of malaria. Therapeutically effective amounts can be determined by monitoring the patient's response to treatment. Typical indications of a therapeutically effective dose include, for example, but are not limited to, amelioration or prevention of malaria symptoms in a patient, including reduction in parasite counts. Amounts effective for this use will depend on the severity of the disease and the general health of the patient, including other factors such as age, weight, sex, and route of administration. Single or multiple administrations of the antibody will depend on the dosage and frequency required and tolerated by the patient.

[0193] In certain embodiments, the antibody is administered at the pre-erythrocytic stage of infection, ie, the antibody is administered in a time frame that prevents or reduces hepatocyte infection.

[0194] Various pharmaceutically acceptable diluents, carriers and excipients, and techniques for preparing and using pharmaceutical compositions are also disclosed herein. Exemplary pharmaceutical compositions and pharmaceutically acceptable diluents, carriers, excipients are also described in Remington: The Science and Practice of Pharmacy 20th Ed. (Lippincott, Williams & Wilkins 2012). In certain embodiments, each carrier, diluent, or excipient is "acceptable" in the sense of being compatible with the other components of the pharmaceutical composition and not harmful to the subject. Often, the pharmaceutically acceptable carrier is a pH buffered aqueous solution. In certain non-limiting embodiments, for example, pharma- ceutically acceptable carriers, diluents, or excipients include water; buffers, e.g., phosphate buffered saline; sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol, polyethylene glycol; esters, such as ethyl oleate, ethyl laurate; agar; buffers, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffers; and other non-toxic compatible substances used in pharmaceutical formulations. Wetting agents, emulsifying agents, and lubricating agents such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring, and perfuming agents, preservatives and antioxidants can also be present in the composition.

[0195] In certain embodiments, the pharmaceutical composition may be formulated for any suitable route of administration, including, for example, parenteral, pulmonary, intranasal, or topical administration. Parenteral administration may include intramuscular, intravenous, intraarterial, intraperitoneal, oral, or subcutaneous administration. In certain embodiments, the pharmaceutical composition is formulated for intravenous administration and has an antibody concentration of 10-100 mg / ml, 10-50 mg / ml, 20-40 mg / ml, or about 30 mg / ml. In certain embodiments, the pharmaceutical composition is formulated for subcutaneous injection and has an antibody concentration of 50-500 mg / ml, 50-250 mg / ml, or 100-150 mg / ml, and a viscosity of less than 50 cP, less than 30 cP, less than 20 cP, or about 10 cP. In certain embodiments, the pharmaceutical composition is a liquid or a solid. In certain embodiments, the pharmaceutical composition is formulated for parenteral administration, e.g., intravenous, subcutaneous, intraperitoneal, or intramuscular administration.

[0196] In certain embodiments, the formulation and delivery method of pharmaceutical composition is adapted according to the site and disease to be treated.For example, but not limited to, the formulation includes antibody is encapsulated in micelles, liposomes, drug release capsules (active agent is incorporated in biocompatible coating designed for slow release); ingestible formulations; topical formulations such as creams, ointments and gels; other formulations such as inhalants, aerosols and sprays.

[0197] In certain non-limiting embodiments, for example, for parenteral administration, the antibody or antigen-binding fragment thereof is formulated into a unit dose injectable form (solution, suspension, emulsion) in association with a pharma- ceutically acceptable parenteral vehicle. Non-limiting examples of vehicles include water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Non-aqueous vehicles such as fixed oils and ethyl oleate can also be used.

[0198] The dosage and administration regimen will depend on a variety of factors, such as the nature of the infection, the characteristics of the subject, and the medical history of the subject, which are readily determined by the physician. In certain embodiments, the amount of the antibody or antigen-binding fragment thereof administered or provided to the subject ranges from about 0.1 mg to about 50 mg per kg of the subject's body weight. Depending on the type and severity of the infection, in certain embodiments, about 0.1 mg to about 50 mg per kg of body weight (e.g., about 0.1-15 mg / kg / administration) of the antibody or antigen-binding fragment thereof may be provided as an initial candidate dosage to the patient, for example, in one or more separate administrations or by continuous infusion. The progress of treatment is readily monitored by conventional methods and assays and based on criteria known to the physician or other skilled artisan.

[0199] The antibody or variant of the present disclosure can be administered to a subject by any route of administration, such as systemic administration, parenteral administration, or local administration, according to known methods. Such routes include, but are not limited to, intravenous administration, for example, as a bolus or by continuous infusion over a period of time, intramuscular, intraperitoneal, intracerebrospinal, subcutaneous, intraarticular, intrasynovial, intrathecal, oral, topical, or inhalation routes. A subject can be administered one or more doses of the antibody of the present invention; and, as further described below, it can be administered before, after, or simultaneously with another therapeutic agent.

[0200] In certain embodiments, the antibody or variants thereof of the present disclosure may be administered to prevent malaria. In certain embodiments, the antibody disclosed herein may inhibit or reduce Plasmodium infection. In certain embodiments, the antibody disclosed herein may inhibit or reduce the pre-erythrocytic or sporozoite stage of infection. In certain embodiments, the antibody disclosed herein may prevent malaria by targeting Plasmodium at the early stage of invasion into the subject vertebrate, thereby preventing infection from occurring.

[0201] In certain embodiments, the disclosed antibodies or variants thereof may be administered to treat malaria. In certain embodiments, the disclosed antibodies may inhibit or reduce the progression of Plasmodium infection in the bloodstream. In certain embodiments, the disclosed antibodies may inhibit or reduce the transmission of Plasmodium from one subject to another via insect feeding, e.g., mosquito bites, or contact with infected blood.

[0202] In certain embodiments, the pharmaceutical compositions disclosed herein may be administered to pediatric patients. The term "pediatric patient" as used herein refers to patients up to 18 years of age. In certain embodiments, pediatric patients are patients between 3 months and under 12 years of age. In certain non-limiting embodiments, pediatric patients may be patients between about 1 and about 2 years of age, about 2 and about 3 years of age, about 3 and about 4 years of age, about 4 and about 5 years of age, about 5 and about 6 years of age, about 6 and about 7 years of age, about 7 and about 8 years of age, about 8 and about 9 years of age, about 9 and about 10 years of age, or about 11 and about 12 years of age. In certain embodiments, pediatric patients are unresponsive or poorly responsive to other treatments for malaria. In certain embodiments, pediatric patients are humans.

[0203] In certain embodiments, the dose of the pharmaceutical composition disclosed herein is administered based on the weight of the pediatric patient.In certain non-limiting embodiments, the dose of the pharmaceutical composition is about 5mg / kg, about 10mg / kg, about 15mg / kg, about 20mg / kg, about 25mg / kg, about 50mg / kg, about 75mg / kg, about 100mg / kg, about 150mg / kg, about 200mg / kg, about 250mg / kg, about 300mg / kg, or about 350mg / kg.In certain embodiments, the pediatric patient has a weight of about 2.5kg to about 5kg, about 5kg to about 10kg, about 10kg to about 15kg, about 15kg to about 20kg, about 20kg to about 30kg, or about 30kg to about 40kg.

[0204] In certain embodiments, the antibody is provided to a subject in combination with one or more additional therapeutic agents used to treat or prevent malaria or related diseases or disorders.In certain embodiments, a method for treating or preventing malaria is provided, comprising administering to a human a therapeutically effective amount of an antibody disclosed herein or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective amount of one or more additional therapeutic agents.In certain embodiments, a method for treating malaria in a human having or at risk of having an infection is provided, comprising administering to a human a therapeutically effective amount of an antibody disclosed herein or a pharmaceutically acceptable salt thereof in combination with a therapeutically effective amount of one or more additional therapeutic agents.

[0205] In certain embodiments, when the antibodies of the present disclosure described herein are combined with one or more additional therapeutic agents as described above, the components of the composition are administered as a simultaneous or sequential regimen. When administered sequentially, the combination may be administered in two or more administrations.

[0206] In certain embodiments, the antibodies disclosed herein are combined with one or more additional therapeutic agents in a unit dosage form for simultaneous administration to a patient.

[0207] "Patient" refers to any subject to which an antibody is administered, whether or not the subject has malaria. In certain embodiments, a "patient" is a non-human subject, e.g., an animal used as a model to evaluate the effects of antibody administration.

[0208] "Co-administration" of an antibody disclosed herein and one or more additional therapeutic agents generally refers to simultaneous or sequential administration of an antibody or fragment thereof disclosed herein and one or more additional therapeutic agents such that therapeutically effective amounts of both the antibody or fragment thereof disclosed herein and the one or more additional therapeutic agents are present in the patient's body. Co-administration includes administering a unit dose of an antibody disclosed herein before or after administration of a unit dose of the one or more additional therapeutic agents, including, but not limited to, administering the antibody within seconds, minutes, or hours of administration of the one or more additional therapeutic agents. In certain non-limiting embodiments, for example, a unit dose of an antibody disclosed herein is administered first, followed by a unit dose of the one or more additional therapeutic agents within seconds or minutes. In certain non-limiting embodiments, a unit dose of the one or more additional therapeutic agents is administered first, followed by a unit dose of the antibody within seconds or minutes. In certain embodiments, a unit dose of an antibody disclosed herein is administered first, followed by a unit dose of the one or more additional therapeutic agents several hours (e.g., 1-12 hours) later. In certain embodiments, a unit dose of the one or more additional therapeutic agents is administered first, followed several hours (eg, 1-12 hours) later by a unit dose of the antibody.

[0209] The administration of the combination may be simultaneous, using separate or a single pharmaceutical composition, or sequential in any order, optionally with a period during which both (or all) therapeutic agents simultaneously exert their biological activity. Such combination therapy may result in a synergistic therapeutic effect. In certain embodiments, it may be desirable to combine the administration of an antibody of the invention with another antibody against another Plasmodium falciparum antigen or another CSP target epitope.

[0210] CSP has multiple domains and regions, including an N-terminal domain, an immunogenic central NANP repeat region, and a C-terminal (ctCSP) domain or an α-thrombospondin repeat (αTSR) domain. Between the N-terminal domain and the central repeat region is a junction region that contains an NPDP sequence, both of which are related to the dominant NANP motif, and a minor repeat region that contains three NVDP motifs (Pholcharee, T. et al., J. Mol. Bio. 432: 1048-1063 (2020)). In certain embodiments, the antibodies disclosed herein are co-administered with an antibody that binds to ctCSP. In certain embodiments, the antibodies disclosed herein are co-administered with an antibody that binds to the alpha epitope (α-ctCSP) domain of ctCSP. α-ctCSP consists of an α-helix that contains the T cell epitope Th2R (region III) and a CS flap that contains another T cell epitope Th3R (Figure 2 in Beutler N, PLoS Pathog 18(3):e1010409(2022), incorporated herein by reference). In certain embodiments, the antibodies disclosed herein are co-administered with an antibody that binds to the beta epitope (β-ctCSP) domain of ctCSP (Figure 2 in Beutler N, PLoS Pathog 18(3):e1010409(2022)).

[0211] In certain embodiments, the antibody may be administered by gene therapy via nucleic acid comprising one or more polynucleotides encoding the antibody. In certain embodiments, the polynucleotide encodes an scFv. In certain embodiments, the polynucleotide comprises DNA, cDNA or RNA. In certain embodiments, the polynucleotide is present in a vector, for example, a viral vector.

[0212] Exemplary embodiments of the subject matter of the present disclosure The present disclosure provides an antibody that targets Plasmodium falciparum. In certain non-limiting embodiments, the antibody is an anti-circumsporozoite protein (CSP) antibody. In certain embodiments, the recombinant antibody comprises a light chain variable region (VL) and a heavy chain variable region (VH), where 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.

[0213] In certain embodiments, the VL of the recombinant antibody disclosed herein comprises the amino acid sequence set forth in SEQ ID NO: 163; the amino acid sequence set forth in SEQ ID NO: 23; the amino acid sequence set forth in SEQ ID NO: 33; the amino acid sequence set forth in SEQ ID NO: 43; the amino acid sequence set forth in SEQ ID NO: 53; the amino acid sequence set forth in SEQ ID NO: 63; the amino acid sequence set forth in SEQ ID NO: 73; the amino acid sequence set forth in SEQ ID NO: 83; the amino acid sequence set forth in SEQ ID NO: 93; the amino acid sequence set forth in SEQ ID NO: 103; the amino acid sequence set forth in SEQ ID NO: 113; the amino acid sequence set forth in SEQ ID NO: 123; the amino acid sequence set forth in SEQ ID NO: 133; the amino acid sequence set forth in SEQ ID NO: 143; the amino acid sequence set forth in SEQ ID NO: 153; or the amino acid sequence set forth in SEQ ID NO: 173. In certain embodiments, the VH of the recombinant antibody disclosed herein comprises the amino acid sequence set forth in SEQ ID NO: 164; the amino acid sequence set forth in SEQ ID NO: 24; the amino acid sequence set forth in SEQ ID NO: 34; the amino acid sequence set forth in SEQ ID NO: 44; the amino acid sequence set forth in SEQ ID NO: 54; the amino acid sequence set forth in SEQ ID NO: 64; the amino acid sequence set forth in SEQ ID NO: 74; the amino acid sequence set forth in SEQ ID NO: 84; the amino acid sequence set forth in SEQ ID NO: 94; the amino acid sequence set forth in SEQ ID NO: 104; the amino acid sequence set forth in SEQ ID NO: 114; the amino acid sequence set forth in SEQ ID NO: 124; the amino acid sequence set forth in SEQ ID NO: 134; the amino acid sequence set forth in SEQ ID NO: 144; the amino acid sequence set forth in SEQ ID NO: 154; or the amino acid sequence set forth in SEQ ID NO: 174.

[0214] In certain embodiments of the recombinant antibodies disclosed herein, 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 amino acid sequence set forth in SEQ ID NO: 84; the VL comprises the amino acid sequence set forth in SEQ ID NO: 93. or wherein the VH comprises the amino acid sequence set forth in SEQ ID NO:94; the VL comprises the amino acid sequence set forth in SEQ ID NO:103 and the VH comprises the amino acid sequence set forth in SEQ ID NO:104; the VL comprises the amino acid sequence set forth in SEQ ID NO:113 and the VH comprises the amino acid sequence set forth in SEQ ID NO:114; the VL comprises the amino acid sequence set forth in SEQ ID NO:123 and the VH comprises the amino acid sequence set forth in SEQ ID NO:124; the VL comprises the amino acid sequence set forth in SEQ ID NO:133 and the VH comprises the amino acid sequence set forth in SEQ ID NO:134; the VL comprises the amino acid sequence set forth in SEQ ID NO:143 and the VH comprises the amino acid sequence set forth in SEQ ID NO:144; the VL comprises the amino acid sequence set forth in SEQ ID NO:153 and the VH comprises the amino acid sequence set forth in SEQ ID NO:154; or the VL comprises the amino acid sequence set forth in SEQ ID NO:173 and the VH comprises the amino acid sequence set forth in SEQ ID NO:174. In certain embodiments of the recombinant antibodies disclosed herein, 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. In certain embodiments of the recombinant antibodies disclosed herein, the VL comprises the amino acid sequence set forth in SEQ ID NO: 133 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 134. In certain embodiments of the recombinant antibodies disclosed herein, 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.

[0215] In certain embodiments, the recombinant antibody disclosed herein comprises a light chain (LC) and a heavy chain (HC). In certain embodiments, the LC of the recombinant antibody disclosed herein 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:167; or the amino acid sequence set forth in SEQ ID NO:177. In certain embodiments, the HC of the recombinant antibody disclosed herein comprises the amino acid sequence set forth in SEQ ID NO:28 or SEQ ID NO:29; the amino acid sequence set forth in SEQ ID NO:38 or SEQ ID NO:39; the amino acid sequence set forth in SEQ ID NO:48 or SEQ ID NO:49; the amino acid sequence set forth in SEQ ID NO:58 or SEQ ID NO:59; the amino acid sequence set forth in SEQ ID NO:68 or SEQ ID NO:69; the amino acid sequence set forth in SEQ ID NO:78 or SEQ ID NO:79; the amino acid sequence set forth in SEQ ID NO:88 or SEQ ID NO:89; the amino acid sequence set forth in SEQ ID NO:98 or SEQ ID NO:99; the amino acid sequence set forth in SEQ ID NO:108 or SEQ ID NO:109; the amino acid sequence set forth in SEQ ID NO:118 or SEQ ID NO:119; the amino acid sequence set forth in SEQ ID NO:128 or SEQ ID NO:129; the amino acid sequence set forth in SEQ ID NO:138 or SEQ ID NO:139; the amino acid sequence set forth in SEQ ID NO:148 or SEQ ID NO:149; the amino acid sequence set forth in SEQ ID NO:158 or SEQ ID NO:159; the amino acid sequence set forth in SEQ ID NO:168 or SEQ ID NO:169; or the amino acid sequence set forth in SEQ ID NO:178 or SEQ ID NO:179.

[0216] In certain embodiments of the recombinant antibodies disclosed herein, the LC comprises the amino acid sequence set forth in SEQ ID NO:27 and the HC comprises the amino acid sequence set forth in SEQ ID NO:28 or SEQ ID NO:29; the LC comprises the amino acid sequence set forth in SEQ ID NO:37 and the HC comprises the amino acid sequence set forth in SEQ ID NO:38 or SEQ ID NO:39; the LC comprises the amino acid sequence set forth in SEQ ID NO:47 and the HC comprises the amino acid sequence set forth in SEQ ID NO:48 or SEQ ID NO:49; the LC comprises the amino acid sequence set forth in SEQ ID NO:57 and the HC comprises the amino acid sequence set forth in SEQ ID NO:58 or SEQ ID NO:59; 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:68 or SEQ ID NO:69; the LC comprises the amino acid sequence set forth in SEQ ID NO:77 and the HC comprises the amino acid sequence set forth in SEQ ID NO:78 or SEQ ID NO:79; the LC comprises the amino acid sequence set forth in SEQ ID NO:87 and the HC comprises the amino acid sequence set forth in SEQ ID NO:88 or SEQ ID NO:89; the LC comprises the amino acid sequence set forth in SEQ ID NO:97 and the HC comprises the amino acid sequence set forth in SEQ ID NO:98 or SEQ ID NO:99; or LC comprises the amino acid sequence set forth in SEQ ID NO: 107 and HC comprises the amino acid sequence set forth in SEQ ID NO: 108 or SEQ ID NO: 109; or LC comprises the amino acid sequence set forth in SEQ ID NO: 117 and HC comprises the amino acid sequence set forth in SEQ ID NO: 118 or SEQ ID NO: 119; or LC comprises the amino acid sequence set forth in SEQ ID NO: 127 and HC comprises the amino acid sequence set forth in SEQ ID NO: 128 or SEQ ID NO: 129; or LC comprises the amino acid sequence set forth in SEQ ID NO: 137 and HC comprises the amino acid sequence set forth in SEQ ID NO: 138 or SEQ ID NO: 139; or LC comprises the amino acid sequence set forth in SEQ ID NO: 147 and HC comprises the amino acid sequence set forth in SEQ ID NO: 148 or SEQ ID NO: 149; or LC comprises the amino acid sequence set forth in SEQ ID NO: 157 and HC comprises the amino acid sequence set forth in SEQ ID NO: 158 or SEQ ID NO: 159; or LC comprises the amino acid sequence set forth in SEQ ID NO: 167 and HC comprises the amino acid sequence set forth in SEQ ID NO: 168 or SEQ ID NO: 169; or LC comprises the amino acid sequence set forth in SEQ ID NO: 177 and HC comprises the amino acid sequence set forth in SEQ ID NO: 178 or SEQ ID NO: 179.In certain embodiments of the recombinant antibodies disclosed herein, 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. In certain embodiments, the LC comprises the amino acid sequence set forth in SEQ ID NO: 137 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 139. In certain embodiments, the LC comprises the amino acid sequence set forth in SEQ ID NO: 167 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 169.

[0217] In certain embodiments, the VL of the recombinant antibodies disclosed herein comprises the amino acid sequence set forth in SEQ ID NO: 13. In certain embodiments of the recombinant antibodies disclosed herein, the VL comprises at least one amino acid substitution. In certain embodiments of the recombinant antibodies disclosed herein, the at least one amino acid substitution is at position 1 and / or position 44. In certain embodiments of the recombinant antibodies disclosed herein, the amino acid substitution at position 1 is E1Q. In certain embodiments of the recombinant antibodies disclosed herein, the amino acid substitution at position 44 is R44T.

[0218] In certain embodiments, the VH of the recombinant antibody disclosed herein comprises the amino acid sequence set forth in SEQ ID NO: 14. In certain embodiments of the recombinant antibody disclosed herein, the VH comprises at least one amino acid substitution. In certain embodiments of the recombinant antibody disclosed herein, the at least one amino acid substitution is at position 21, 23, 88, 98, or a combination thereof. In certain embodiments of the recombinant antibody disclosed herein, the amino acid substitution at position 1 is E1Q. In certain embodiments of the recombinant antibody disclosed herein, the amino acid substitution at position 44 is R44T. In certain embodiments of the recombinant antibody disclosed herein, the amino acid substitution at position 21 is P21S. In certain embodiments of the recombinant antibody disclosed herein, the amino acid substitution at position 23 is T23A. In certain embodiments of the recombinant antibody disclosed herein, the amino acid substitution at position 80 is I80T. In certain embodiments, the amino acid substitution at position 90 is T90A.

[0219] In certain embodiments, the HC of the recombinant antibodies disclosed herein comprises the amino acid sequence set forth in SEQ ID NO: 18. In certain embodiments of the recombinant antibodies disclosed herein, the HC comprises at least one amino acid substitution. In certain embodiments of the recombinant antibodies disclosed herein, the at least 438 amino acid substitutions are at positions 1 and / or 444. In certain embodiments, the amino acid substitution at position 438 is M438L. In certain embodiments of the recombinant antibodies disclosed herein, the amino acid substitution at position 444 is N444S.

[0220] In certain non-limiting embodiments, the present disclosure provides a recombinant anti-circumsporozoite protein (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. In certain embodiments, 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.

[0221] In certain non-limiting embodiments, the disclosure further provides a recombinant anti-circumsporozoite protein (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: 133 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 134. In certain embodiments, 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: 137 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 139.

[0222] In certain non-limiting embodiments, the present disclosure also provides a recombinant anti-circumsporozoite protein (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: 163 and the VH comprises the amino acid sequence set forth in SEQ ID NO: 164. In certain embodiments, 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: 167 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 169.

[0223] In certain non-limiting embodiments, the present disclosure provides a recombinant anti-circumsporozoite protein (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: 183, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 184, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 185; and the heavy chain variable region comprises a CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 186, a CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 187, and a CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 188. In certain embodiments, the VL comprises the amino acid sequence set forth in SEQ ID NO: 195, and the VH comprises the amino acid sequence set forth in SEQ ID NO: 196. In certain embodiments, the recombinant antibody comprises a LC and a HC. In certain embodiments of the recombinant antibody disclosed herein, the LC comprises the amino acid sequence set forth in SEQ ID NO: 199, and the HC comprises the amino acid sequence set forth in SEQ ID NO: 200 or SEQ ID NO: 201.

[0224] In certain non-limiting embodiments, the present disclosure provides a recombinant anti-circumsporozoite protein (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.

[0225] In certain embodiments, the VL of a recombinant antibody disclosed herein comprises the amino acid sequence set forth in SEQ ID NO: 227; the amino acid sequence set forth in SEQ ID NO: 237; the amino acid sequence set forth in SEQ ID NO: 247; the amino acid sequence set forth in SEQ ID NO: 257; or the amino acid sequence set forth in SEQ ID NO: 267. In certain embodiments, the VH of a recombinant antibody disclosed herein comprises the amino acid sequence set forth in SEQ ID NO: 228; the amino acid sequence set forth in SEQ ID NO: 238; the amino acid sequence set forth in SEQ ID NO: 248; the amino acid sequence set forth in SEQ ID NO: 258; or the amino acid sequence set forth in SEQ ID NO: 268. In certain embodiments of the recombinant antibodies disclosed herein, the VL comprises the amino acid sequence set forth in SEQ ID NO:227 and the VH comprises the amino acid sequence set forth in SEQ ID NO:228; the VL comprises the amino acid sequence set forth in SEQ ID NO:237 and the VH comprises the amino acid sequence set forth in SEQ ID NO:238; the VL comprises the amino acid sequence set forth in SEQ ID NO:247 and the VH comprises the amino acid sequence set forth in SEQ ID NO:248; the VL comprises the amino acid sequence set forth in SEQ ID NO:257 and the VH comprises the amino acid sequence set forth in SEQ ID NO:258; or the VL comprises the amino acid sequence set forth in SEQ ID NO:267 and the VH comprises the amino acid sequence set forth in SEQ ID NO:268.

[0226] In certain embodiments, the recombinant antibody disclosed herein comprises an LC and an HC. In certain embodiments, the LC of the recombinant antibody disclosed herein comprises the amino acid sequence set forth in SEQ ID NO:231; the amino acid sequence set forth in SEQ ID NO:241; the amino acid sequence set forth in SEQ ID NO:251; the amino acid sequence set forth in SEQ ID NO:261; or the amino acid sequence set forth in SEQ ID NO:271. In certain embodiments, the HC of the recombinant antibody disclosed herein comprises the amino acid sequence set forth in SEQ ID NO:232 or SEQ ID NO:233; the amino acid sequence set forth in SEQ ID NO:242 or SEQ ID NO:243; the amino acid sequence set forth in SEQ ID NO:252 or SEQ ID NO:253; the amino acid sequence set forth in SEQ ID NO:262 or SEQ ID NO:263; or the amino acid sequence set forth in SEQ ID NO:272 or SEQ ID NO:273. In certain embodiments of the recombinant antibodies disclosed herein, the LC comprises the amino acid sequence set forth in SEQ ID NO:231 and the HC comprises the amino acid sequence set forth in SEQ ID NO:232 or SEQ ID NO:233; the LC comprises the amino acid sequence set forth in SEQ ID NO:241 and the HC comprises the amino acid sequence set forth in SEQ ID NO:242 or SEQ ID NO:243; the LC comprises the amino acid sequence set forth in SEQ ID NO:251 and the HC comprises the amino acid sequence set forth in SEQ ID NO:252 or SEQ ID NO:253; the LC comprises the amino acid sequence set forth in SEQ ID NO:261 and the HC comprises the amino acid sequence set forth in SEQ ID NO:262 or SEQ ID NO:263; or the LC comprises the amino acid sequence set forth in SEQ ID NO:271 and the HC comprises the amino acid sequence set forth in SEQ ID NO:272 or SEQ ID NO:273.

[0227] In certain embodiments, the VH of the recombinant antibodies disclosed herein comprises the amino acid sequence set forth in SEQ ID NO: 218. In certain embodiments of the recombinant antibodies disclosed herein, the VH comprises at least one amino acid substitution. In certain embodiments of the recombinant antibodies disclosed herein, the at least one amino acid substitution is at position 40, 69, 80, 85, 120, or a combination thereof. In certain embodiments of the recombinant antibodies disclosed herein, the amino acid substitution at position 40 is T40A. In certain embodiments of the recombinant antibodies disclosed herein, the amino acid substitution at position 69 is I69T. In certain embodiments of the recombinant antibodies disclosed herein, the amino acid substitution at position 80 is S80Y. In certain embodiments of the recombinant antibodies disclosed herein, the amino acid substitution at position 85 is G85S. In certain embodiments of the recombinant antibodies disclosed herein, the amino acid substitution at position 120 is I120T. In certain embodiments of the recombinant antibodies disclosed herein, the HC comprises the amino acid sequence set forth in SEQ ID NO: 222. In certain embodiments of the recombinant antibodies disclosed herein, the HC comprises at least one amino acid substitution. In certain embodiments of the recombinant antibodies disclosed herein, the at least 434 amino acid substitutions are at positions 1 and / or 440. In certain embodiments of the recombinant antibodies disclosed herein, the amino acid substitution at position 434 is M434L. In certain embodiments of the recombinant antibodies disclosed herein, the amino acid substitution at position 440 is N440S.

[0228] In certain embodiments, the recombinant antibodies disclosed herein exhibit at least a 20% reduction in parasite liver burden compared to the reference antibody. In certain embodiments, the recombinant antibodies disclosed herein exhibit at least a 20% increase in survival compared to the reference antibody. In certain embodiments, the recombinant antibodies disclosed herein exhibit increased conformational stability compared to the reference antibody. In certain embodiments, the recombinant antibodies disclosed herein exhibit increased colloidal stability compared to the reference antibody. In certain embodiments, the reference antibody is AB-000317. In certain embodiments, the reference antibody is AB-000224. In certain embodiments, the reference antibody is AB-007088.

[0229] In certain embodiments, the recombinant antibodies disclosed herein bind to the NANP repeat region. In certain embodiments, the recombinant antibodies disclosed herein bind to a polypeptide comprising or consisting of the amino acid sequence set forth in SEQ ID NO:280.

[0230] In certain embodiments, a recombinant antibody disclosed herein comprises at least one modification to the native AB-000224 variable heavy chain amino acid sequence set forth in SEQ ID NO: 14. In certain embodiments, a recombinant antibody comprises at least one modification to the native AB-000224 variable light chain amino acid sequence set forth in SEQ ID NO: 13. In certain embodiments, a recombinant antibody disclosed herein comprises at least one modification to the native AB-000224 variable heavy chain amino acid sequence set forth in SEQ ID NO: 14 and at least one modification to the native AB-000224 variable light chain amino acid sequence set forth in SEQ ID NO: 13.

[0231] In certain embodiments, the recombinant antibody disclosed herein comprises at least one modification to the native AB-007088 variable heavy chain amino acid sequence set forth in SEQ ID NO: 218. In certain embodiments, the recombinant antibody comprises at least one modification to the native AB-007088 variable light chain amino acid sequence set forth in SEQ ID NO: 217. In certain embodiments, the recombinant antibody disclosed herein comprises at least one modification to the native AB-007088 variable heavy chain amino acid sequence set forth in SEQ ID NO: 218 and at least one modification to the native AB-000224 variable light chain amino acid sequence set forth in SEQ ID NO: 217. In certain non-limiting embodiments, the present disclosure provides a polynucleotide encoding the recombinant antibody disclosed herein. In certain non-limiting embodiments, the present disclosure provides an expression vector comprising the polynucleotide disclosed herein. In certain non-limiting embodiments, the present disclosure provides a host cell comprising the expression vector or polynucleotide disclosed herein.

[0232] In certain non-limiting embodiments, the present disclosure provides a composition comprising a recombinant antibody disclosed herein. In certain embodiments, the composition further comprises a pharma- ceutically acceptable carrier.

[0233] In certain non-limiting embodiments, the present disclosure provides a method of preventing and / or treating malaria in a subject in need thereof comprising administering an effective amount of a recombinant antibody disclosed herein or a composition disclosed herein. In certain embodiments, the subject is a pediatric patient.

[0234] In certain non-limiting embodiments, the present disclosure provides a recombinant antibody or composition disclosed herein for use in the prevention and / or treatment and / or prophylaxis of malaria in a subject in need of treatment. Additionally, in certain non-limiting embodiments, the present disclosure provides a recombinant antibody or composition disclosed herein for the manufacture of a medicament for the prevention and / or treatment and / or prophylaxis of malaria in a subject in need of treatment. Furthermore, the present disclosure provides the use of a recombinant antibody or composition disclosed herein for the prevention and / or treatment and / or prophylaxis of malaria in a subject in need of treatment. In certain embodiments, the subject is a pediatric patient.

[0235] From the foregoing description, it will be apparent that variations and modifications may be made to the subject matter disclosed herein to adapt it to various uses and conditions, and such embodiments also fall within the scope of the following claims.

[0236] The recitation of a list of elements in a definition of a variable herein includes definition of that variable as any single element or combination (or subcombination) of the listed elements. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

[0237] All patents and publications mentioned in this specification are herein incorporated by reference to the same extent as if each individual patent and publication was specifically and individually indicated to be incorporated by reference.

[0238] All features disclosed herein may be combined in any combination. Each feature disclosed herein may be replaced by an alternative feature serving the same, equivalent, or similar purpose. Thus, unless expressly stated otherwise, each feature disclosed is only an example of a generic series of equivalent or similar features.

[0239] The foregoing written description is believed to be sufficient to enable one of ordinary skill in the art to practice the methods and / or obtain the compositions described herein. The following examples and detailed description are offered by way of illustration and not by way of limitation. EXAMPLES

[0240] Working Example The examples are provided for illustrative purposes only and are not intended to limit the scope of the invention in any way. Indeed, various modifications in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and fall within the scope of the appended claims.

[0241] Given the above general description, it will be appreciated that various other embodiments may be practiced.

[0242] Example 1. Identification of functionally active anti-CSP antibodies AB-000224 and AB-007088 were discovered from antibody repertoires generated by Immune Repertoire Capture® (IRC®) technology from plasmablast B cells isolated from two donors enrolled in a Phase 2a trial evaluating the efficacy of the RTS,S vaccine in preventing malaria infection. The IRC® technology and its use in antibody discovery are well known and are disclosed, for example, in WO2012148497A2, the contents of which are incorporated herein by reference in their entirety. The RTS,S vaccine is a pseudovirion vaccine that combines the central repeat and C-terminal regions of the Hepatitis B surface antigen and CSP proteins. RTS,S consists of two polypeptides; RTS is a single polypeptide chain corresponding to amino acids 207 to 395 of P. falciparum (3D7) fused to HBsAg, and S is a 226 amino acid polypeptide corresponding to HBsAg. Stoute,et.al.,N Engl J Med;336:86-91(1997);RTS,S Clinical Trials Partnership,PLoS Med.11(7:e1001685,(2014),WO1993 / 10152.To enhance efficacy, the RTS,S vaccine was administered with the adjuvant AS01B. AS01B is a liposome-based formulation containing the immunostimulants monophosphoryl lipid A (MPL) and QS21, which has been shown to be more immunogenic than another adjuvant used in earlier trials, AS02A.Kester,et al.,J Infect Dis 200:337-346 (2009). All study participants received one of two vaccine schedules (standard full dose: 0, 1, 2M, or split third dose: 0, 1, 7M) or placebo and were then challenged with a controlled human malaria parasite infection. Donors identified as AB-000224 and AB-007088 were protected after challenge. The heavy and light chain sequences of AB-000224 were expressed as a human IgG1 monoclonal antibody. The heavy and light chain sequences of AB-007088 were expressed as a human IgG1 monoclonal antibody.AB-000224 and AB-007088 showed stronger binding and affinity to CSP proteins in vitro, did not bind to Hepatitis B proteins, and showed superior functional activity when tested in vivo compared to other antibodies obtained from the same or different donors.

[0243] This example provides the design of improved variants of AB-000224 or AB-007088. In certain embodiments, the variants produced have improved exploitability, as identified by various in vitro assays, such as aggregation assessment by HPLC or UPLC, hydrophobic interaction chromatography (HIC), multispecific assays (e.g., baculovirus particle binding), self-interaction nanoparticle spectroscopy (SINS), or mass spectrometry after incubation under accelerated degradation conditions, such as high temperature, low pH, high pH, ​​or oxidizing H2O2. If activity is maintained (or enhanced), while the severity of the disadvantage is eliminated or reduced, the mutation is successful.

[0244] Example 2. Generation of anti-CSP antibody variants Evaluation of AB-000224 In vivo generated antibodies undergo somatic hypermutation following genomic recombination. Germline information was obtained from the AB-000224 antibody and used to optimize variant design. Figures 4A and 4B show the alignment of AB-0002224 to the putative V and J germline genes. CDRs, germline deviations, and potential detrimental motifs were identified. N-linked glycosylation sites, atypical cysteine ​​residues, and other potential detrimental motifs were identified across VH and VL.

[0245] Closely related siblings of AB-000224 include AB-007110, AB-007111, and AB-007112. These were evaluated at the sequence and structure levels to find potentially beneficial modifications to AB-000224. Notably, AB-007110 was shown to have improved thermal stability over AB-000224 in biophysical characterization. The positions evaluated below are distinct from AB-000224 and are either consistent between siblings or structurally interesting.

[0246] Design of variants of AB-000224 The deviations from germline in the framework and complementarity determining regions (CDRs) in AB-000224 were analyzed for the possibility of mutating the germline sequence, alone or in combination, without adversely affecting binding or potency to the (NANP)3 region of the CSP protein. For each of the candidate mutations from the sequence of AB-000224 to the germline sequence, the risk of making the mutation was evaluated based on: (1) the change in charge, if any, since charge changes are inherently risky, and changes to a more positive charge are especially risky, given that the net charge of the AB-000224 Fv is already positive; (2) the conservation of the native AB-000224 residue in the sequence and the presence of germline residues or other mutations at that position in the sequence, and (3) the structural position of that position relative to the NANP motif. It was noted that some mutations were linked to at least one other mutation, meaning that the risk prediction was based on making the mutation in conjunction with the other mutation(s). The proposed AB-000224 residue modifications are shown below in Table 31 according to the ASN numbering system:

[0247] [Table 31]

[0248] Mutations were constructed by grouping the "standard" design group in all combinations. This resulted in 16 variants. The "sibling" design group was used to add one variant grouping three sites together. This resulted in a total of 17 variants. The addition of the parents provided 18 antibodies for generation. The locations of the mutation sites in the AB-000224 variants are shown in the table below according to the ASN numbering system:

[0249] [Table 32]

[0250] Evaluation of AB-007088 Germline information was obtained from the AB-007088 antibody and used to optimize variant design. Figures 5A and 5B show the alignment of AB-0070884 to the putative V and J germline genes. CDRs, germline deviations, and potential detrimental motifs were identified. N-linked glycosylation sites, atypical cysteine ​​residues, and other potential detrimental motifs were identified across VH and VL.

[0251] Design of variants of AB-007088 The deviations from germline in the framework and complementarity determining regions (CDRs) in AB-007088 were analyzed for the possibility of mutating the germline sequence, alone or in combination, without adversely affecting binding or potency to the (NANP)3 region of the CSP protein. For each of the candidate mutations from the sequence of AB-007088 to the germline sequence, the risk of making the mutation was evaluated based on: (1) the change in charge, if any, since charge changes are inherently risky, and changes to a more positive charge are especially risky, given that the net charge of the AB-007088 Fv is already positive; (2) the conservation of the native AB-007088 residue in the sequence and the presence of germline residues or other mutations at that position in the sequence, and (3) the structural position of that position relative to the NANP motif. It was noted that some mutations were linked to at least one other mutation, meaning that the risk prediction was based on making the mutation in conjunction with the other mutation(s). The proposed AB-007088 residue modifications are shown below in Table 33 according to the ASN numbering system:

[0252] [Table 33]

[0253] Mutations were constructed by grouping the "Group 1" design groups in all combinations. This resulted in four variants. A single variant with HV:S90Y was added to the designs with all mutations, resulting in a total of five designs. The positions of the mutation sites in the AB-007088 variants are shown in the table below according to the ASN numbering system:

[0254] [Table 34]

[0255] Example 3. Binding Assays AB-000224 and AB-007088 were evaluated for binding to the complete CSP protein and a series of linear peptides representing the immunodominant NANP repeat region. Two assay platforms were used to quantify antibody-target binding: Biolayer Interferometry (BLI) and Surface Plasmon Resonance (SPR). Five binding targets were evaluated in the SPR assay and six targets were evaluated in the BLI platform and are listed in Table 35 below:

[0256] [Table 35]

[0257] For BLI, each target shown in Table 35 was biotinylated and immobilized on a streptavidin sensor. Each antibody was evaluated in duplicate at 5 μg / mL. Antibody-target measurements were repeated if the variation between the two duplicates exceeded 3-fold.

[0258] For SPR, each antibody was either directly coupled to a Carterra Chip or coupled using a goat anti-human Fc antibody. Uncoupled antibodies were washed away and various concentration gradients of target were run over the antibodies, where the highest concentration of each target ranged from 0.5 to 8 μg / mL. To allow for duplicate measurements, each antibody was immobilized at two different locations on the chip. Affinity for each antibody-target combination was determined with Mathematica software using four to five different target concentrations. Antibody-target measurements were repeated if variation between the two duplicates exceeded three-fold.

[0259] The data generated by the BLI and SPR assays are similar, but the assays were designed with the target and antibody orientations reversed. Specifically, in the BLI assay, the target is fixed and the antibody flows over it, whereas in the SPR assay, the antibody is fixed and the target flows over it. Given these orientations, the antibody is more likely to be involved in binding interactions involving multiple target molecules when assessed in the BLI assay. Thus, antibody-target binding in the BLI assay may show more similarity to binding with the complete CSP protein that covers the surface of malaria sporozoites. In contrast, the activity measured in the SPR assay will more accurately represent the interaction of the antibody F(ab) with a single target molecule. The data generated for the AB-000224 and AB-007088 antibodies are summarized in Table 36 below:

[0260] [Table 36] JPEG2024524895000080.jpg55153

[0261] Overview of AB-000224 antibody variants Seventeen variants (AB-000224.001, AB-000224.002, AB-000224.003, AB-000224.004, AB-000224.005, AB-000224.006, AB-000224.007, AB-000224.009, AB-000224.001, AB-000224.002, AB-000224.003, AB-000224.004, AB-000224.005, AB-000224.006, AB-000224.007, AB-000224.008, AB-000224.009, AB-000224.001 We designed the following: AB-000224.007, AB-000224.008, AB-000224.009, AB-000224.010, AB-000224.011, AB-000224.012, AB-000224.013, AB-000224.014, AB-000224.015, AB-000224.016, AB-000224.017).

[0262] Overview of AB-007088 antibody variants Five variants (AB-007088.001, AB-007088.002, AB-007088.003, AB-007088.004, AB-007088.005) were designed to reduce the risk of antibody-induced immunogenicity by mutating residues in either the framework regions or CDRs into the germline antibody.

[0263] Example 4. In vivo performance of antibodies Liver loading assay AB-000224 and AB-007088 and their variants (with heavy chain version 2 included, designated "LS") were evaluated for in vivo activity in a mouse malaria liver burden assay as described in Flores-Garcia Y, et al. Malar J. 2019;18(1):426, doi:10.1186 / s12936-019-3055-9. Experimental antibodies were compared to positive (AB-000317) and negative (AB-001245) antibody controls. AB-000317 is an anti-CSP antibody described in WO2020 / 172220. AB-001245 is a non-CSP isotype control.

[0264] For each antibody, five C57B1 / 6 mice per experimental or control group were administered 100 μg of antibody 16 hours before intravenous infection with fluorescent chimeric P. berghei sporozoites expressing the CSP protein of P. falciparum. Forty-two hours after parasite challenge, liver burden of sporozoites was quantified by bioluminescence. Percent liver burden was calculated for each experimental mouse by subtracting the mean background luminescence measured in two untreated naive mice and calculating the percent reduction compared to the mean luminescence measured in five untreated infected mice. The mean percent reduction was reported for each of the experimental antibody groups.

[0265] Mice treated with AB-000224, AB-007088, or the positive control antibody AB-000317 all showed similar reductions in liver burden compared to naive infected mice and mice treated with the negative control AB-001245 (Figure 1A).Levels of experimental and control human antibodies circulating in mice upon infection were quantified by ELISA assay (Figure 1B) and were similar between experimental and control antibodies.

[0266] The AB-000224 and AB-007088 variants resulted in reduced liver burden in treated mice to the same extent as the respective parental antibodies AB-000224 or AB-007088 and the positive control antibody AB-000317. Liver burden levels measured in animals administered either the variants, parent molecule or the positive control antibody AB-000317 were significantly lower than liver burden levels in either 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. Concentrations of human antibodies circulating in mice at the time of infection were measured by ELISA and are shown in Figures 10G-10I.

[0267] Bite-wound parasitemia assay Protective capacity was also evaluated for AB-000224 and AB-007088 and variants thereof (forms containing heavy chain version 2 are designated "LS"). In this experiment, animals were exposed to mosquitoes infected with chimeric P. berghei expressing the P. falciparum CSP protein as described in Espinosa, D., et al. npj Vaccines 2017;2,10(2017); Espinosa, D., et al. Infect Immun. 2013 Aug;81(8):2882-2887.

[0268] C57Bl / 6 mice were administered 150 μg of antibody and exposed 16 h later to six or seven chimeric P. berghei-infected mosquitoes. At least 70%-80% of the mosquito population was infected with chimeric P. berghei expressing the CSP protein of P. falciparum, resulting in exposure to at least one infection event. Each mouse was then assessed for blood-stage parasitemia 4–10 days post-infection by microscopy. Parasitemia data were recorded as positive or negative and used to generate survival curves. Experimental antibodies were compared to both a positive control (AB-000317) and a negative control (AB-001245). Mice treated with AB-000224, AB-007088, or the positive control AB-000317 were less likely to develop parasitemia than mice treated with the negative control AB-001245. (Figures 2A and 2C show experimental data for AB-000224, and Figures 3A and 3C show experimental data for AB-007088.) Levels of circulating human antibodies in mice during infection were quantified by ELISA assay (Figures 2B and 3B) and were similar between the experimental and control antibodies.

[0269] Finally, mice administered AB-000224 or AB-007088 variants were as likely to remain parasite-free as mice treated with either the parental antibody or the positive control antibody AB-000317 (Figures 11A-11C). The concentration of human antibodies circulating in mice at the time of infection was measured by ELISA and is shown in Figures 11D-11F.

[0270] Example 5. Biophysical characterization This example describes the biophysical characterization of anti-CSP antibody variants. Conformational and colloidal stability was evaluated for AB-000224 and AB-007088 and their variants (all in heavy chain version 2 format). Several methods were used to analyze these endpoints and are summarized in Tables 37 and 38 below:

[0271] [Table 37]

[0272] [Table 38]

[0273] AB-000224 variants showed improved stability AB-000224 and its variants showed comparable yield and titer results. Therefore, additional endpoints were evaluated. The thermal stability of AB-000224 and its variants was assessed by differential scanning fluorimetry (DSF). Thermal unfolding was monitored by measuring the intensity of an external dye (Sypro Orange) as samples were heated from 20°C to 90°C. The data are reported as Tm1 and Tm2, ​​with the first transition correlating with the CH2 domain and the second with the unfolding of the Fab and CH3 domain regions. Higher unfolding temperatures are desirable and are associated with increased conformational stability of the product. The absence of Tm2 indicates that the Fab unfolds at the same or similar temperature as the CH2 domain, reported as Tm1. Further information is obtained from a unique parameter called the weighted shoulder score, which takes into account multiple pieces of information available from the unfolding curves. Again, higher values ​​indicate greater conformational stability. DSF assays were performed in PBS buffer and all samples were diluted to a final antibody concentration of 0.15 mg / mL.

[0274] The tendency to aggregate at elevated temperatures was assessed using a heat aggregation method. Samples were placed in a 96-well Biorad PCR plate and heated to various temperatures for 5 min using a Biorad Thermal Cycler. After heating, protein precipitation was measured by reading the absorbance at 350 nm (A350) using a Spectrostar nanoplate reader. Nearly all AB-000224 variants showed higher WSS compared to the parent antibody AB-000224, had T2 approaching 80 °C, and did not produce precipitation during the heat hold assay. However, AB-000224.017 did not have a T2 and showed precipitation during the heat hold assay.

[0275] Additionally, the chemical stability of AB-000224 and its variants was also evaluated. Sensitivity to low pH was assessed by titrating the samples to pH 3.3 with acetic acid and holding for 30 min, followed by neutralization of the samples to pH 5 with Tris base and measuring aggregation by size-exclusion HPLC. Samples diluted in PBS with the same amount of acetic acid and Tris base as the test samples were used as controls. All AB-000224 and its variants did not show low pH instability.

[0276] AB-000224 and its variants were tested for stability against chemical unfolding as assayed by denaturation with guanidine. Chemical unfolding curves were generated by exposing the antibody to increasing concentrations of guanidine hydrochloride. After 24 hours, the intrinsic fluorescence of the samples was measured using a SUPR-UV plate reader. The collected raw data was then processed and the chemical unfolding curves and their inflection points were calculated from the processed data as a function of denaturant conditions. Antibodies with denaturation inflection points higher than 2.1M guanidine are considered conformationally stable by this method. AB-000224 and variants all showed inflection points above 2M guanidine (Gdn). In particular, AB-000224.005, AB-000224.008, AB-000224.010, AB-000224.011, AB-000224.013, and AB-000224.015 have improved stability against chemical unfolding compared to AB-000224, each having an inflection point above 2.3 M guanidine.

[0277] The colloidal stability of AB-000224 and its variants was also evaluated by self-interaction nanoparticle spectroscopy (SINS), which monitors protein-protein interactions by entrapment on gold colloidal surfaces and measuring the shift in maximum absorption wavelength. Maximum absorption values ​​at wavelengths higher than 550 nm are considered interactive and may cause increased viscosity and filterability problems. All variants had maximum absorption values ​​below 550 nm, showing slight improvements in SINS values ​​compared to AB-000224.

[0278] AB-000224 and its variants were tested for potential hydrophobic interactions that could lead to manufacturing issues by monitoring retention times on a Zenix HPLC column. Undiluted samples were loaded onto the Zenix column and eluted isocratically with a running buffer of 100 mM sodium phosphate, pH 7.0 and monitored at 220 nm. Longer retention times are indicative of hydrophobic interactions. Most antibodies have retention times between 8.5 and 9.0 minutes under the conditions tested. AB-00224 and all its variants showed comparable retention times of approximately 10 minutes.

[0279] The solubility of AB-000224 and its variants was assayed by precipitating antibody samples with increasing amounts of PEG 10,000, filtering the samples, and measuring the concentration of soluble protein. For each parent molecule, initial experiments were performed to determine the ideal PEG concentration that would precipitate approximately 50% of the parent molecule, and subsequent experiments used this PEG concentration for all variants to assess whether the variants were more or less soluble than the parent molecule. Antibodies that precipitate with 8-10% PEG or more are considered highly soluble, while less soluble antibodies precipitate with 4-5% PEG. AB-000224 and all of its variants demonstrated high solubility.

[0280] The polyreactivity of AB-000224 and its variants was determined by testing the antibodies by Elisa for binding to KLH, insulin and dsDNA. Samples were diluted to 1 μg / mL and a secondary anti-human antibody was used to detect the amount of protein bound to the different antigens. After addition of substrate, absorbance was measured at 405 nm. As a positive control, polyreactive antibodies were used. Nonspecific binding to such common physiological components can cause an increase in clearance rate and negatively affect the pK. Absorbances above 1.5 for KLH and insulin and above 2.0 for dsDNA may indicate problems with nonspecific binding to the molecule. Most AB-000224 variants did not show polyreactivity, but several variants showed polyreactive signals to insulin (AB-000224.006, AB-000224.007, AB-000224.008, AB-000224.009, AB-000224.010).

[0281] AB-000224 and its variants were ranked based on the results of the different assays and potencies, as shown in Figures 12B and 12C.

[0282] AB-007088 variants showed improved stability The yield and potency of AB-007088 and its variants were first evaluated. The variants of AB-007088 showed comparable yield and potency results. AB-007088 and its variants were evaluated to determine conformational and colloidal stability as described above for AB-000224 and its variants.

[0283] The variants of AB-007088.005 showed improved thermal stability by the DFS method, with T2 approaching 75°C, whereas the other variants were similar to the parent AB-007088, with T1 values ​​of approximately 70°C and T2 not measurable. In addition, AB-007088.005 showed slightly reduced precipitation compared to the parent AB-007088 and other variants. Chemical stability was evaluated, and AB-007088 and all of its variants showed no instability at low pH (pH 3.3). Furthermore, all of AB-007088 and its variants showed stability against chemical unfolding, with inflection points at or above 2M guanidine (Gdn). Of note, AB-007088.001 showed improved stability compared to AB-007088, with inflection points at or above 2.3M guanidine.

[0284] The colloidal stability of AB-007088 and its variants was also evaluated. Protein-protein interactions were monitored (SINS), and all variants showed very low SINS values, indicating the absence of protein-protein interactions. In addition, AB-007088 and its variants showed comparable retention times on the Zenix column. Of note, PEG solubility was measured, and AB-007088 and all its variants showed high solubility. Furthermore, most of the variants of AB-007088 did not show polyreactivity, although some variants showed polyreactivity signals to insulin (AB-007088.001, AB-007088.003, AB-007088.004).

[0285] AB-007088 and its variants were ranked based on the results of the different assays and potencies, as shown in Figures 13B and 13C.

Claims

**Claim 1**: An antibody comprising a light chain variable region (VL) and a heavy chain variable region (VH), a) the light chain variable region comprises CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 2, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 3; and the heavy chain variable region comprises CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 4, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 5, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 6; b) the light chain variable region comprises CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 183, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 184, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 185; and the heavy chain variable region comprises CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 186, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 187, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 188; or c) the light chain variable region comprises CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 205, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 206, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 207; and the heavy chain variable region comprises CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 208, CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 209, and CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 210, a recombinant anti - circumsporozoite protein (CSP) antibody. **Claim 2**: wherein VL is a) the amino acid sequence set forth in SEQ ID NO: 163; b) the amino acid sequence set forth in SEQ ID NO: 23; c) the amino acid sequence set forth in SEQ ID NO: 33; d) the amino acid sequence set forth in SEQ ID NO: 43; e) the amino acid sequence set forth in SEQ ID NO: 53; f) the amino acid sequence set forth in SEQ ID NO: 63; g) the amino acid sequence set forth in SEQ ID NO: 73; h) the amino acid sequence set forth in SEQ ID NO: 83; i) the amino acid sequence set forth in SEQ ID NO: 93; j) the amino acid sequence set forth in SEQ ID NO: 103; k) the amino acid sequence set forth in SEQ ID NO: 113; l) the amino acid sequence set forth in SEQ ID NO: 123; m) the amino acid sequence set forth in SEQ ID NO: 133; n) the amino acid sequence set forth in SEQ ID NO: 143; o) the amino acid sequence set forth in SEQ ID NO: 153; p) the amino acid sequence set forth in SEQ ID NO: 173; q) the amino acid sequence set forth in SEQ ID NO: 195; r) the amino acid sequence set forth in SEQ ID NO: 227; s) the amino acid sequence set forth in SEQ ID NO: 237; t) the amino acid sequence set forth in SEQ ID NO: 247; u) the amino acid sequence set forth in SEQ ID NO: 257; or v) the amino acid sequence set forth in SEQ ID NO: 267 and / or VH is a) the amino acid sequence set forth in SEQ ID NO: 164; b) the amino acid sequence set forth in SEQ ID NO: 24; c) the amino acid sequence set forth in SEQ ID NO: 34; d) the amino acid sequence set forth in SEQ ID NO: 44; e) the amino acid sequence set forth in SEQ ID NO: 54; f) the amino acid sequence set forth in SEQ ID NO: 64; g) the amino acid sequence set forth in SEQ ID NO: 74; h) the amino acid sequence set forth in SEQ ID NO: 84; i) the amino acid sequence set forth in SEQ ID NO: 94; j) the amino acid sequence set forth in SEQ ID NO: 104; k) the amino acid sequence set forth in SEQ ID NO: 114; l) the amino acid sequence set forth in SEQ ID NO: 124; m) the amino acid sequence set forth in SEQ ID NO: 134; n) the amino acid sequence set forth in SEQ ID NO: 144; (o) the amino acid sequence set forth in SEQ ID NO: 154; (p) the amino acid sequence set forth in SEQ ID NO: 174; (q) the amino acid sequence set forth in SEQ ID NO: 196; (r) the amino acid sequence set forth in SEQ ID NO: 228; (s) the amino acid sequence set forth in SEQ ID NO: 238; (t) the amino acid sequence set forth in SEQ ID NO: 248; (u) the amino acid sequence set forth in SEQ ID NO: 258; or (v) the amino acid sequence set forth in SEQ ID NO: 268 The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 1, comprising:

3. (a) VL comprises the amino acid sequence set forth in SEQ ID NO: 163, and VH comprises the amino acid sequence set forth in SEQ ID NO: 164; (b) VL comprises the amino acid sequence set forth in SEQ ID NO: 23, and VH comprises the amino acid sequence set forth in SEQ ID NO: 24; (c) VL comprises the amino acid sequence set forth in SEQ ID NO: 33, and VH comprises the amino acid sequence set forth in SEQ ID NO: 34; (d) VL comprises the amino acid sequence set forth in SEQ ID NO: 43, and VH comprises the amino acid sequence set forth in SEQ ID NO: 44; (e) VL comprises the amino acid sequence set forth in SEQ ID NO: 53, and VH comprises the amino acid sequence set forth in SEQ ID NO: 54; (f) VL comprises the amino acid sequence set forth in SEQ ID NO: 63, and VH comprises the amino acid sequence set forth in SEQ ID NO: 64; (g) VL comprises the amino acid sequence set forth in SEQ ID NO: 73, and VH comprises the amino acid sequence set forth in SEQ ID NO: 74; (h) VL comprises the amino acid sequence set forth in SEQ ID NO: 83, and VH comprises the amino acid sequence set forth in SEQ ID NO: 84; (i) VL comprises the amino acid sequence set forth in SEQ ID NO: 93, and VH comprises the amino acid sequence set forth in SEQ ID NO: 94; (j) VL comprises the amino acid sequence set forth in SEQ ID NO: 103, and VH comprises the amino acid sequence set forth in SEQ ID NO: 104; k) VL contains the amino acid sequence set forth in SEQ ID NO: 113 and VH contains the amino acid sequence set forth in SEQ ID NO: 114; l) VL contains the amino acid sequence set forth in SEQ ID NO: 123 and VH contains the amino acid sequence set forth in SEQ ID NO: 124; m) VL contains the amino acid sequence set forth in SEQ ID NO: 133 and VH contains the amino acid sequence set forth in SEQ ID NO: 134; n) VL contains the amino acid sequence set forth in SEQ ID NO: 143 and VH contains the amino acid sequence set forth in SEQ ID NO: 144; o) VL contains the amino acid sequence set forth in SEQ ID NO: 153 and VH contains the amino acid sequence set forth in SEQ ID NO: 154; p) VL contains the amino acid sequence set forth in SEQ ID NO: 173 and VH contains the amino acid sequence set forth in SEQ ID NO: 174; q) VL contains the amino acid sequence set forth in SEQ ID NO: 195 and VH contains the amino acid sequence set forth in SEQ ID NO: 196; r) VL contains the amino acid sequence set forth in SEQ ID NO: 227 and VH contains the amino acid sequence set forth in SEQ ID NO: 228; s) VL contains the amino acid sequence set forth in SEQ ID NO: 237 and VH contains the amino acid sequence set forth in SEQ ID NO: 238; t) VL contains the amino acid sequence set forth in SEQ ID NO: 247 and VH contains the amino acid sequence set forth in SEQ ID NO: 248; u) VL contains the amino acid sequence set forth in SEQ ID NO: 257 and VH contains the amino acid sequence set forth in SEQ ID NO: 258; or v) VL contains the amino acid sequence set forth in SEQ ID NO: 267 and VH contains the amino acid sequence set forth in SEQ ID NO: 268, The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 1.

4. The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 1, wherein VL contains the amino acid sequence set forth in SEQ ID NO: 163 and VH contains the amino acid sequence set forth in SEQ ID NO:

164.

5. Comprising a light chain (LC) and a heavy chain (HC), wherein LC is a) the amino acid sequence set forth in SEQ ID NO: 27; b) the amino acid sequence set forth in SEQ ID NO: 37; c) the amino acid sequence set forth in SEQ ID NO: 47; d) the amino acid sequence set forth in SEQ ID NO: 57; e) the amino acid sequence set forth in SEQ ID NO: 67; f) the amino acid sequence set forth in SEQ ID NO: 77; g) the amino acid sequence set forth in SEQ ID NO: 87; h) the amino acid sequence set forth in SEQ ID NO: 97; i) the amino acid sequence set forth in SEQ ID NO: 107; j) the amino acid sequence set forth in SEQ ID NO: 117; k) the amino acid sequence set forth in SEQ ID NO: 127; l) the amino acid sequence set forth in SEQ ID NO: 137; m) the amino acid sequence set forth in SEQ ID NO: 147; n) the amino acid sequence set forth in SEQ ID NO: 157; o) the amino acid sequence set forth in SEQ ID NO: 167; p) the amino acid sequence set forth in SEQ ID NO: 177; q) the amino acid sequence set forth in SEQ ID NO: 199; r) the amino acid sequence set forth in SEQ ID NO: 231; s) the amino acid sequence set forth in SEQ ID NO: 241; t) the amino acid sequence set forth in SEQ ID NO: 251; u) the amino acid sequence set forth in SEQ ID NO: 261; or v) the amino acid sequence set forth in SEQ ID NO: 271 and / or HC is a) the amino acid sequence set forth in SEQ ID NO: 28 or SEQ ID NO: 29; b) the amino acid sequence set forth in SEQ ID NO: 38 or SEQ ID NO: 39; c) the amino acid sequence set forth in SEQ ID NO: 48 or SEQ ID NO: 49; d) the amino acid sequence set forth in SEQ ID NO: 58 or SEQ ID NO: 59; e) the amino acid sequence set forth in SEQ ID NO: 68 or SEQ ID NO: 69; (f) the amino acid sequence set forth in SEQ ID NO: 78 or SEQ ID NO: 79; (g) the amino acid sequence set forth in SEQ ID NO: 88 or SEQ ID NO: 89; (h) the amino acid sequence set forth in SEQ ID NO: 98 or SEQ ID NO: 99; (i) the amino acid sequence set forth in SEQ ID NO: 108 or SEQ ID NO: 109; (j) the amino acid sequence set forth in SEQ ID NO: 118 or SEQ ID NO: 119; (k) the amino acid sequence set forth in SEQ ID NO: 128 or SEQ ID NO: 129; (l) the amino acid sequence set forth in SEQ ID NO: 138 or SEQ ID NO: 139; (m) the amino acid sequence set forth in SEQ ID NO: 148 or SEQ ID NO: 149; (n) the amino acid sequence set forth in SEQ ID NO: 158 or SEQ ID NO: 159; (o) the amino acid sequence set forth in SEQ ID NO: 168 or SEQ ID NO: 169; (p) the amino acid sequence set forth in SEQ ID NO: 178 or SEQ ID NO: 179; (q) the amino acid sequence set forth in SEQ ID NO: 200 or SEQ ID NO: 201; (r) the amino acid sequence set forth in SEQ ID NO: 232 or SEQ ID NO: 233; (s) the amino acid sequence set forth in SEQ ID NO: 242 or SEQ ID NO: 243; (t) the amino acid sequence set forth in SEQ ID NO: 252 or SEQ ID NO: 253; (u) the amino acid sequence set forth in SEQ ID NO: 262 or SEQ ID NO: 263; or (v) the amino acid sequence set forth in SEQ ID NO: 272 or SEQ ID NO: 273 The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 1, comprising

6. (a) LC comprises the amino acid sequence set forth in SEQ ID NO: 27, and HC comprises the amino acid sequence set forth in SEQ ID NO: 28 or SEQ ID NO: 29; (b) LC comprises the amino acid sequence set forth in SEQ ID NO: 37, and HC comprises the amino acid sequence set forth in SEQ ID NO: 38 or SEQ ID NO: 39; (c) The LC contains the amino acid sequence set forth in SEQ ID NO: 47, and the HC contains the amino acid sequence set forth in SEQ ID NO: 48 or SEQ ID NO: 49; (d) The LC contains the amino acid sequence set forth in SEQ ID NO: 57, and the HC contains the amino acid sequence set forth in SEQ ID NO: 58 or SEQ ID NO: 59; (e) The LC contains the amino acid sequence set forth in SEQ ID NO: 67, and the HC contains the amino acid sequence set forth in SEQ ID NO: 68 or SEQ ID NO: 69; (f) The LC contains the amino acid sequence set forth in SEQ ID NO: 77, and the HC contains the amino acid sequence set forth in SEQ ID NO: 78 or SEQ ID NO: 79; (g) The LC contains the amino acid sequence set forth in SEQ ID NO: 87, and the HC contains the amino acid sequence set forth in SEQ ID NO: 88 or SEQ ID NO: 89; (h) The LC contains the amino acid sequence set forth in SEQ ID NO: 97, and the HC contains the amino acid sequence set forth in SEQ ID NO: 98 or SEQ ID NO: 99; (i) The LC contains the amino acid sequence set forth in SEQ ID NO: 107, and the HC contains the amino acid sequence set forth in SEQ ID NO: 108 or SEQ ID NO: 109; (j) The LC contains the amino acid sequence set forth in SEQ ID NO: 117, and the HC contains the amino acid sequence set forth in SEQ ID NO: 118 or SEQ ID NO: 119; (k) The LC contains the amino acid sequence set forth in SEQ ID NO: 127, and the HC contains the amino acid sequence set forth in SEQ ID NO: 128 or SEQ ID NO: 129; (l) The LC contains the amino acid sequence set forth in SEQ ID NO: 137, and the HC contains the amino acid sequence set forth in SEQ ID NO: 138 or SEQ ID NO: 139; (m) The LC contains the amino acid sequence set forth in SEQ ID NO: 147, and the HC contains the amino acid sequence set forth in SEQ ID NO: 148 or SEQ ID NO: 149; (n) The LC contains the amino acid sequence set forth in SEQ ID NO: 157, and the HC contains the amino acid sequence set forth in SEQ ID NO: 158 or SEQ ID NO: 159; (o) The LC contains the amino acid sequence set forth in SEQ ID NO: 167, and the HC contains the amino acid sequence set forth in SEQ ID NO: 168 or SEQ ID NO: 169; (p) The LC contains the amino acid sequence set forth in SEQ ID NO: 177, and the HC contains the amino acid sequence set forth in SEQ ID NO: 178 or SEQ ID NO:

179. (q) The LC contains the amino acid sequence set forth in SEQ ID NO: 199, and the HC contains the amino acid sequence set forth in SEQ ID NO: 200 or SEQ ID NO: 201; (r) The LC contains the amino acid sequence set forth in SEQ ID NO: 231, and the HC contains the amino acid sequence set forth in SEQ ID NO: 232 or SEQ ID NO: 233; (s) The LC contains the amino acid sequence set forth in SEQ ID NO: 241, and the HC contains the amino acid sequence set forth in SEQ ID NO: 242 or SEQ ID NO: 243; (t) The LC contains the amino acid sequence set forth in SEQ ID NO: 251, and the HC contains the amino acid sequence set forth in SEQ ID NO: 252 or SEQ ID NO: 253; (u) The LC contains the amino acid sequence set forth in SEQ ID NO: 261, and the HC contains the amino acid sequence set forth in SEQ ID NO: 262 or SEQ ID NO: 263; or (v) The LC contains the amino acid sequence set forth in SEQ ID NO: 271, and the HC contains the amino acid sequence set forth in SEQ ID NO: 272 or SEQ ID NO: 273, The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 5.

7. The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 6, wherein the LC contains the amino acid sequence set forth in SEQ ID NO: 167 and the HC contains the amino acid sequence set forth in SEQ ID NO:

169.

8. A recombinant anti-sporozoite circumsporozoite protein (CSP) antibody comprising a light chain variable region (VL) and a heavy chain variable region (VH), wherein the VL contains an amino acid sequence having at least one amino acid substitution with respect to the sequence set forth in SEQ ID NO: 13, at least one amino acid substitution in the VL is at position 1 and / or position 44, and / or the VH contains an amino acid sequence having at least one amino acid substitution with respect to the sequence set forth in SEQ ID NO:

14.

9. The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 8, wherein the amino acid substitution at position 1 is E1Q and the amino acid substitution at position 44 is R44T. **Claim 10**: The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 9, wherein at least one amino acid substitution in VH is at position 21, 23, 88, 98, or a combination thereof. **Claim 11**: The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 10, wherein the amino acid substitution at position 21 is P21S, the amino acid substitution at position 23 is T23A; the amino acid substitution at position 80 is I80T; and the amino acid substitution at position 90 is T90A. **Claim 12**: The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 8, wherein at least one amino acid substitution in the VH region is at position 80, 21, 99, 90, or a combination thereof. **Claim 13**: The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 12, wherein the amino acid substitution at position 80 is I80T, the amino acid substitution at position 21 is P21S, the amino acid substitution at position 99 is T99A, and the amino acid substitution at position 90 is T90A. **Claim 14**: The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 1, wherein the antibody comprises a heavy chain (HC) comprising the amino acid sequence set forth in SEQ ID NO: 18, and at least one amino acid substitution is at position 438 and / or 444. **Claim 15**: The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 14, wherein the amino acid substitution at position 438 is M438L and the amino acid substitution at position 444 is N444S. **Claim 16**: A recombinant anti-sporozoite circumsporozoite protein (CSP) antibody comprising a light chain variable region (VL) and a heavy chain variable region (VH), a) wherein VL comprises the amino acid sequence set forth in SEQ ID NO: 163 and VH comprises the amino acid sequence set forth in SEQ ID NO: 164; b) wherein VL comprises the amino acid sequence set forth in SEQ ID NO: 63 and VH comprises the amino acid sequence set forth in SEQ ID NO: 64; or c) wherein VL comprises the amino acid sequence set forth in SEQ ID NO: 133 and VH comprises the amino acid sequence set forth in SEQ ID NO:

134. **Claim 20**: A recombinant anti-sporozoite circumsporozoite protein (CSP) antibody. **Claim 17** Comprising a light chain (LC) and a heavy chain (HC), a) the LC comprises the amino acid sequence set forth in SEQ ID NO: 167 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 168 or 169; b) 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; or c) the LC comprises the amino acid sequence set forth in SEQ ID NO: 137 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 139, The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 16. **Claim 18** Comprising a light chain (LC) and a heavy chain (HC), a) 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; b) the LC comprises the amino acid sequence set forth in SEQ ID NO: 137 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 139; or c) the LC comprises the amino acid sequence set forth in SEQ ID NO: 167 and the HC comprises the amino acid sequence set forth in SEQ ID NO: 168 or SEQ ID NO: 169, A recombinant anti-sporozoite circumsporozoite protein (CSP) antibody. **Claim 19** The VH comprises the amino acid sequence set forth in SEQ ID NO: 218 and the VH comprises at least one amino acid substitution at position 40, 69, 80, 85, 120, or a combination thereof, the recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 1. **Claim 20** a) the amino acid substitution at position 40 is T40A, b) the amino acid substitution at position 69 is I69T, c) the amino acid substitution at position 80 is S80Y, d) the amino acid substitution at position 85 is G85S, and e) the amino acid substitution at position 120 is I120T, The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 19. **Claim 21** The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 1, wherein HC comprises the amino acid sequence set forth in SEQ ID NO: 222 and comprises at least one amino acid substitution at position 434 and / or position 440. **Claim 22** The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to claim 21, wherein the amino acid substitution at position 434 is M434L and the amino acid substitution at position 440 is N440S. **Claim 23** The antibody a) shows at least a 20% reduction in parasite liver load compared to a reference antibody, b) shows at least a 20% increase in survival rate compared to a reference antibody, c) shows an increase in conformational stability compared to a reference antibody, and / or d) shows an increase in colloidal stability compared to a reference antibody and the reference antibody is selected from the group consisting of AB-000317, AB-000224, and AB-007088, the recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to any one of claims 1 to 22. **Claim 24** The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to any one of claims 1 to 22, wherein the antibody binds to the NANP repeat region. **Claim 25** The recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to any one of claims 1 to 22, wherein the antibody binds to a polypeptide comprising or consisting of the amino acid sequence set forth in SEQ ID NO:

280. **Claim 26** A polynucleotide encoding the recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to any one of claims 1 to 22. **Claim 27** An expression vector comprising the polynucleotide according to claim 26. **Claim 28** A host cell comprising the polynucleotide according to claim 26 or an expression vector comprising said polynucleotide. **Claim 29** A composition comprising the recombinant anti-sporozoite circumsporozoite protein (CSP) antibody according to any one of claims 1 to 22. The composition according to claim 29, further comprising a pharmaceutically acceptable carrier. The composition according to claim 29 for use in the prevention and / or treatment of malaria in a subject in need of treatment, optionally wherein the subject is a pediatric patient.