| 主权项 |
1. A nucleic acid comprising or consisting of a sequence encoding for the light chain of a mutated humanized 12G4 monoclonal antibody and/or a sequence encoding for the heavy chain of said mutated humanized 12G4 monoclonal antibody, said mutated humanized 12G4 monoclonal antibody comprising or consisting of:
a) a light chain comprising or consisting of: a variable region (VL) according to amino acid sequence SEQ ID NO: 2 or SEQ ID NO: 4, said VL consisting of three regions determining antigen recognition (CDR) surrounded by four framework regions (FR), and a constant region according to amino acid sequence SEQ ID NO: 6 or a sequence having at least 80% homology with SEQ ID NO: 6 , and b) a heavy chain comprising or consisting of: a variable region (VH) according to amino acid sequence SEQ ID NO: 8 or SEQ ID NO: 10, and a constant region according to amino acid sequence SEQ ID NO: 12 or a sequence having at least 80% homology with SEQ ID NO: 12,wherein said mutated humanized 12G4 monoclonal antibody comprises at least one mutation selected from the group consisting of:
a mutation located in one of the CDR regions of the VL that is a substitution of at least one amino acid selected from the group consisting of S179P, one of E184K, E184G, or E184D, and S182F, a mutation located in one of the FR regions of VL that is a substitution of at least one amino acid selected from the group consisting of: I177T, I132T, A143T, T150A, S158P, L175Q, Y178H, V187A, S192T, G197D, and F212S, and a mutation located in the heavy chain that is a substitution of at least one amino acid selected from the group consisting of: Q1E, Q3E or Q3R, Q6E, A9T, V11A, K12R, K13R, K19E, V20A, one of A24G, A24V or A24T, Q39E, A40V, S31G, L45P, D56N, A76T, A79T, R87G, T58A, Q62R, V67M, I70N, T74A, S77P, A79T, S88P, E89D, F102S, A1031, L110P, and S114T, andsaid mutated humanized 12G4 monoclonal antibody has a KD for human anti-Müllerian hormone type II receptor (AMHRII) at least equal to that of the chimeric 12G4 monoclonal antibody, comprising or consisting of:
a) a light chain consisting of: a variable region according to amino acid sequence SEQ ID NO: 14, and a constant region according to amino acid sequence SEQ ID NO: 6, b) a heavy chain consisting of: a variable region according to amino acid sequence SEQ ID NO: 18, and a constant region according to amino acid sequence SEQ ID NO: 12, for said receptor below 10−7 M. |
