Plant-derived antibodies and derivatives that reduce risk of antibody-dependent enhancement (ADE) of infection
The present invention describes the plant-based production of a therapeutic anti-virus antibody.
1. A ΔXF N. benthamiana plant-produced monoclonal antibody (MAb) specific for West Nile Virus (WNV), the MAb comprising a defined and highly-uniform N-glycoform, wherein the N-glycoform is Man5, Man7, Man8 and/or Man9;
wherein the MAb comprises pHu-E16scFv-CH1-3, wherein the MAb lacks a KDEL sequence,
wherein the MAb has reduced antibody-dependent enhancement (ADE) as compared to a mammalian-produced MAb,
wherein the MAb has equivalent antigen binding affinity and kinetics, neutralization activity, and in vivo therapeutic activity against a target virus infection as compared to a corresponding antibody produced from non-plant cells, and wherein the MAb comprises a heavy chain (HC) further comprising one or more amino acid mutations that affect Fc receptor or/and C1q binding, wherein the mutations are located at L234, L235 and/or G237 in a human IgG1 isotype HC, or at equivalent locations that correspond to, or align with these positions in a different MAb isotype or subtype, wherein L234, L235 and G237 are equivalent to L255, L256 and G258, respectively, in SEQ ID NO:34.
2. The plant-produced MAb of claim 1 , wherein the N-glycoform selectively binds to Fc receptors or C1q.
3. The plant-produced MAb of claim 1 , wherein the MAb is pHu-E16scFv-CH1-3.
4. The plant-produced MAb of claim 1 , wherein the MAb is Tetra pHu-E16.
5. The plant-produced MAb of claim 1 , wherein the HC is a human IgG1 isotype and the mutations are selected from L234AL235A and L234AL235AG237A, or equivalent mutations that correspond to, or align with these positions.
6. The plant-produced MAb of claim 1 , wherein the N-glycoform is Man7 and/or Man8.
7. A ΔXF N. benthamiana plant-produced monoclonal antibody (MAb) specific for dengue virus (DENV), the MAb comprising a defined and highly-uniform N-glycoform, wherein the N-glycoform is GnGnXF, GnGn or oligomannosidic;
wherein the MAb is pE60 or a single chain variant of DV MAb E60 (scFv-Fc fusion molecule), wherein the MAb lacks a KDEL sequence,
wherein the MAb has reduced antibody-dependent enhancement (ADE) as compared to a mammalian-produced MAb,
wherein the MAb has equivalent antigen binding affinity and kinetics, neutralization activity, and in vivo therapeutic activity against a target virus infection as compared to a corresponding antibody produced from non-plant cells,
wherein the MAb comprises a heavy chain (HC) further comprising one or more amino acid mutations that affect Fc receptor or/and C1q binding, wherein the mutations are located at L234, L235 and/or G237 in a human IgG1 isotype HC, or at equivalent locations that correspond to, or align with these positions in a different MAb isotype or subtype, wherein L234, L235 and G237 are equivalent to L255, L256 and G258, respectively, in SEQ ID NO:34.
8. The plant-produced MAb of claim 7 , wherein the HC comprises two or three amino acid mutations.
9. The plant-produced MAb of claim 8 , wherein the mutations are L234A and L235A or L234A, L235A and G237A, or equivalent mutations that correspond to, or align with these positions.
10. The plant-produced Mab of claim 7 , wherein the N-glycoform is GnGn.
11. The plant-produced Mab of claim 7 , wherein the N-glycoform is GnGnXF.
12. The plant-produced Mab of claim 7 , wherein the N-glycoform has an oligomannosidic structure.
13. The plant-produced MAb of claim 12 , wherein the N-glycoform is Man5, Man7, Man8, and/or Man9.
14. The plant-produced MAb of claim 7 , wherein the MAb comprises:
1) a heavy chain comprising SEQ ID NO:18, SEQ ID NO:22 or SEQ ID NO:24;
2) a heavy chain, wherein the heavy chain comprises an amino acid sequence encoded by SEQ ID NO:5, SEQ ID NO:17, SEQ ID NO:21 or SEQ ID NO:23;
3) a light chain, wherein the light chain comprises an amino acid sequence encoded by SEQ ID NO:4;
4) an amino acid sequence encoded by SEQ ID NO:6, SEQ ID NO:25, SEQ ID NO:29 or SEQ ID NO:31; and/or
5) an amino acid sequence comprising SEQ ID NO:26, SEQ ID NO:30 or SEQ ID NO:32.
15. The plant-produced MAb of claim 7 , wherein the MAb comprises a light chain, the light chain comprising an alanine-serine (AS) peptide between the variable and constant domains of the light chain.
16. The plant-produced MAb of claim 1 , wherein the MAb comprises an amino acid sequence encoded by SEQ ID NO:3.