scFv and Antibodies with Reduced Multimerisation
AssigneeY-Mabs Therapeutics, Inc.
InventorsTorben LUND-HANSEN, Nico LIEBENBERG, Matias Munck MORTENSEN, Steen LISBY
Life of this patent
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Abstract
Disclosed are methods for generating variants of scFv, having reduced tendency of forming multimers.
Claims82
A bispecific antibody comprising a first scFv domain capable of binding a chelator or a chelator binding a metal ion, a second scFv domain capable of binding a tumor antigen, and a SADA domain, wherein the VH and VL domains in the first scFv and/or the second scFv domain is/are not connected by a disulfide bond
The bispecific antibody according to any of the preceding claims, wherein the first scFv domain capable of binding a chelator or a chelator binding a metal ion, is selected among scFvs capable of binding DOTA, a derivative of DOTA, DOTAM or any of these binding a metal ion.
The bispecific antibody according to any of the preceding claims, wherein the tumor antigen is selected among:
- HER2, B7-H3, CA6, CD138, CD20, CD19, CD22, CD27L, CD30, CD33, CD37, CD38, CD47, CD56, CD66e, CD70, CD74, CD79b, EGFR, EGFRvIII, FRα, GCC, GPNMB, Mesothelin, MUC16, NaPi2b, Nectin 4, PSMA, STEAP1, Trop-2, 5T4, AGS-16, alpha v beta6, CA19.9, CAIX, CD138, CD174, CD180, CD227, CD326, CD79a, CEACAM5, CRIPTO, DLL3, DS6, Endothelin B receptor, FAP, GD2, Mesothelin, PMEL 17, SLC44A4, TENB2, TIM-1, CD98, Endosialin/CD248/TEM1, Fibronectin Extra-domain B, LIV-1, Mucin 1, p-cadherin, peritosin, Fyn, SLTRK6, Tenascin c, VEGFR2, and PRLR.
The bispecific antibody of claim 61, comprising
- a. a VL sequence with the sequence of SEQ ID NO:
- 56 or a sequence having at least 90% sequence identity, e.g., at least 95% sequence identity, e.g., at least 96% sequence identity, e.g., at least 97% sequence identity, e.g., at least 98% sequence identity or at least 99% sequence identity to SEQ ID NO:
- 56, wherein the amino acid in position 44 is not a cysteine;
- and
- b. a VH sequence with the sequence of SEQ ID NO:
- 57 or a sequence having at least 90% sequence identity, e.g., at least 95% sequence identity, e.g., at least 96% sequence identity, e.g., at least 97% sequence identity, e.g., at least 98% sequence identity or at least 99% sequence identity to SEQ ID NO:
- 57, wherein the amino acid in position 100 is not a cysteine.
The bispecific antibody of claim 61 or 62, wherein the second scFv comprises or consists of the sequence of SEQ ID NO:
- 61, or comprising or consisting of a sequence having at least 90% sequence identity, e.g., at least 95% sequence identity, e.g., at least 96% sequence identity, e.g., at least 97% sequence identity, e.g., at least 98% sequence identity or at least 99% sequence identity to SEQ ID NO:
- 61.
The bispecific antibody according to any of the preceding claims, wherein the SADA domain is selected among domains comprising one of the sequences of SEQ ID NO:
- 5-12 or sequences that differs from one of SEQ ID NO:
- 5-12 by 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 substitutions.
The bispecific antibody according to any of the preceding claims, wherein the SADA domain comprises an amino acid sequence of amino acids 6-36 of SEQ ID NO:
- 5 or a sequence that differs from amino acids 6-36 of SEQ ID NO:
- 5 by one or more substitutions selected among:
- E6V, Q, K, G, D or A;
- Y7S, N, H, F, D or C;
- F8Y, V, S, L, I or C;
- T9S, P, N or A;
- L10V, I or F;
- Q11R, L, K, H or E;
- I12V, T, M, L or F;
- R13S, P, L, H, G or C;
- G14W, R or A;
- R15S, P, L, H, G or C;
- E16V, Q, K, G, D or A;
- F18Y, V, S, L, I or C;
- E19V, Q, K, G, D or A;
- M20V, T, R, L, K or I;
- F21L or I;
- R22L or G;
- E23V, Q, K, G, D or A;
- L24M;
- N25S, I or D;
- E26V, Q, K, G, D or A;
- A27V, T, S, G or D;
- L28W, V, M or F;
- E29Q, G or D;
- L30V, R, I, H or F;
- K31T, R, Q, N, M or E;
- D32Y, V, N, H, G or A;
- A33V, T, S, P, G or D;
- Q34R, L, K, H or E;
- using the numbering of SEQ ID NO:
- 5.
The bispecific antibody according to any of the preceding claims, comprising or consisting of one of the sequences of SEQ ID NO:
- 40, 41, 42, 58, 59, 62, 63, 64, 65, 66, 67, 68 and 69.
A method of generating variants of a scFv domain, comprising a light chain variable domain (VL), a heavy chain variable domain (VH) and one or more disulfide bonds between the VL and the VH, comprising the steps of
- a. Identifying the cysteine residues forming said one or more disulfide bonds between VL and VH;
- and
- b. Substituting the cysteine residues forming one or more of said disulfide bonds identified in step a., with amino acids different from cysteine.
Dependencies are inferred from the claim text.