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Humanized transformation method of antibody

A technology of humanization and antibody, which is applied in the direction of chemical instruments and methods, peptides, specific peptides, etc., can solve problems such as inability to complete humanization steps, reduce the time for repeated modeling and expression trial and error, and reduce molecular construction and cell expression, the effect of saving the total cost

Inactive Publication Date: 2019-04-16
SHANGHAI WUXI BIOLOGIC TECH CO LTD +2
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] As far as the existing technology is concerned, during the process of antibody humanization transformation, there are still irreplaceable amino acids in the key regions of some antibodies, and the final humanization step cannot be completed
Existing technology is difficult to effectively solve this problem

Method used

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  • Humanized transformation method of antibody
  • Humanized transformation method of antibody
  • Humanized transformation method of antibody

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Embodiment Construction

[0032] The technical solution of the present invention will be clearly and completely described below, obviously, the described embodiments are part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

[0033] Unless otherwise stated, the implementation of the later examples of this project in the present invention will use conventional molecular biology and biochemistry experimental methods, and these methods are known to those skilled in the art, practitioners or scholars. These techniques are fully described in the following documents, for example: Michael R. Green, Joseph Sambrook, "Molecular Cloning Experiment Guide" 4th Edition Book 1-3 (2017); relevant commercial reagents should be provided by the reagent manufacturer Operate according t...

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Abstract

The invention discloses a humanized transformation method of an antibody. According to the method, purified IgG antibody protein in a humanized transformation process of the antibody is treated by N-glycosidase, wherein N-glycosidase is a PNGaseF enzyme, the sequence Asn-X-Ser / Thr in the protein is specifically recognized by the PNGaseF enzyme, glycosylation is completely removed while disulfide bonds in the antibody protein are maintained, and X represents any amino acid except proline. The invention also provides specific application of the method. The method can be used for effectively replacing some unreplaceable amino acids which still exist in a part of antibodies in the later stage of humanized antibody transformation in the prior art. Through the method, appropriate human sequencescan be reserved as many as possible after screening, the purpose of reducing immune reactions is achieved, and meanwhile, the specificity and affinity of the human sequences can be reserved to the maximum extent, so that a reasonable humanized effect is achieved.

Description

technical field [0001] The present invention belongs to the research field of antibody humanization design in molecular biology, especially relates to the use of N-glycosidase to remove N-glycosyl groups that may affect the conformation of antibody complementarity-determining regions, so as to maintain the singleness and purity of antibody molecules . Background technique [0002] At present, antibody humanization design, as a new research and development trend, has become an important link in the process of today's therapeutic antibody research and development. [0003] Antibody humanization: it is a complex process, mainly through molecular biology experiments, supplemented by computer prediction and simulation of antibody protein structure, so that the mouse monoclonal antibody sequence is transformed into a sequence as close as possible to the human antibody sequence. A method of molecular biology. The antibody successfully modified by this method not only retains the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00
CPCC07K16/00C07K2317/24
Inventor 高晨阳陆晓峰雷鸣王卓智李竞陈智胜
Owner SHANGHAI WUXI BIOLOGIC TECH CO LTD
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