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In vitro molecular directed evolution method for reshaping antibody

A technology of molecular targeting and antibodies, applied in biochemical equipment and methods, using vectors to introduce foreign genetic material, botanical equipment and methods, etc., can solve the problems of verifying the degree of humanization of transplanted antibodies and modified antibodies

Inactive Publication Date: 2005-01-19
BEIJING ABT GENETIC ENG TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the limitations of experimental methods, there are no experimental results to verify the degree of humanization of transplanted antibodies and reshaped antibodies

Method used

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  • In vitro molecular directed evolution method for reshaping antibody
  • In vitro molecular directed evolution method for reshaping antibody
  • In vitro molecular directed evolution method for reshaping antibody

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Embodiment

[0092] 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. The experimental route of the molecular directed evolution method is shown in Fig. 1 .

[0093] 1. Molecular design of primary reshaped anti-4-1BB single chain antibody library

[0094] (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), acco...

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Abstract

The invention relates to an in vitro molecular directed evolution method for reshaping antibody, which integrates ribosome displaying technology with DNA reorganization technology by designing reshaped antibody molecules, thus directly obtaining the extracorporal molecules of the single-chain antibody reshaped orientation.

Description

technical field [0001] The present invention relates to an in vitro molecular directed evolution method for antibody reshaping, more specifically, the present invention relates to the development of murine single-chain antibody by designing reshaped antibody molecules and combining ribosome display technology and DNA shuffling technology. An in vitro molecular directed evolution method for antibody reshaping based on direct access to reshaped single-chain antibodies. The present invention also provides the amino acid sequence and its encoding nucleotide sequence of an anti-4-1BB modified antibody obtained by this method, the vector containing the nucleotide sequence and the host cell containing these vectors. Background technique [0002] Compared with polyclonal antibodies, monoclonal antibodies have good specificity and higher affinity; they can be mass-produced with reproducibility; It is widely used in research and other fields. However, during application, especially ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/09C12N15/12C12N15/63C12P21/00
Inventor 王祥武黄华樑周炳林晴
Owner BEIJING ABT GENETIC ENG TECH
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