Heterodimeric FC (fragment crystallizable) modification method based on charge network and preparation method of heterodimeric proteins

Abstract

The invention discloses a heterodimeric FC (fragment crystallizable) modification method based on a charge network and a preparation method of heterodimeric proteins. The modification method is characterized by establishing a charge interaction network and mutating one or more interfacial amino acids based on interaction of heavy-chain CH3 two-arm amino acids of the FC and modifying the amino acids in the CH3 domains through charge effect, thus weakening self interactions of the domains (which is beneficial to forming homodimers) and enhancing the interactions between the domains ( which is beneficial to forming heterodimers) and finally selecting sense mutation. The invention also relates to heterodimeric proteins prepared by the preparation method, independent polypeptides forming the heterodimeric FC and nucleotide sequences coding the polypeptides.

Description

technical field [0001] The present invention relates to a method for preparing proteins and polypeptides related to the FC of the heterodimeric antibody, and also relates to the FC protein of the heterodimeric antibody and the polypeptide itself, including the individual polypeptides constituting the FC of the heterodimeric antibody; the present invention also relates to Nucleic acid sequences encoding these polypeptides, and pharmaceutical compositions comprising one or more heterogeneous FC proteins or polypeptides. technical background [0002] Monoclonal antibody drugs have grown rapidly in the past fifteen years and have become a growth point in the pharmaceutical industry. Since 1996, a total of about 30 monoclonal antibody drugs have been approved for marketing. Among them, nine monoclonal antibody drugs have annual sales exceeding one billion US dollars. In 2010, the total sales of monoclonal antibody drugs exceeded 30 billion US dollars, and the annual growth rate...

AI Technical Summary

Problems solved by technology

There are two defects in the inventive method. One is that the technical solution lacks completeness. The interaction between interface amino acids is not limited to a pair of amino acids, and the interface amino acids are replaced with opposite charges, especially in the case of multiple amino acid mutations. , based on the complex interactions between interfacial amino acids, electrostatic interactions do not always inhibit homodimers or favor heterodimer formation, although the patent includes some specific substitutions that do favor heterodimers However, there is no strategy or method for evaluation and screening in its technical solution. In the case of complex interface amino acid interactions or multiple point mutations, this patent cannot guide those skilled in the art in order to obtain meaningful mutations; The second is that the technical solution is limited to the replacement of charged amino acids with opposite electrical properties. In fact, a charge interaction network is formed between interface amino acids, and any change in the electrical properties of an amino acid (charged or uncharged amino acid) will cause two polypeptide chains Changes in the electrostatic interaction between charged amino acids are limited to oppositely charged amino acids, which ignores the large number of possible sense mutations

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