In vitro molecular directed evolution method for reshaping antibody
A molecular orientation, antibody technology, applied in biochemical equipment and methods, introduction of foreign genetic material using vectors, botanical equipment and methods, etc., can solve problems such as verifying the degree of humanization of transplanted antibodies and modified antibodies
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[0091] According to the evolutionary conservation of antibody structure and amino acid sequence, a degenerate modified anti-4-1BB single-chain antibody library was designed, and then ribosome display technology and DNA shuffling technology were combined to realize molecular directed evolution in vitro. The reshaped anti-4-1BB single chain antibody was screened from the reshaped single chain antibody library. For the experimental route of this molecular directed evolution method, see figure 1 .
[0092] 1. Molecular design of primary reshaped anti-4-1BB single chain antibody library
[0093] (1) The mouse with agonistic activity reported in the literature (Garni-Wagner BA, et al., 4-1BB is expressed on CD45RAhiROhi transitional T cell in humans. Cell Immunol. 1996 Apr 10; 169 (1): 91-8.) The amino acid sequences of the heavy chain variable region (VH) and light chain variable region (VL) of the anti-4-1BB antibody (4b4-1 clone) (see Figure 2.1M4B4VH and Figure 2.2M4B4VL), acc...
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