High efficiency CHO cell display-based antibody maturation

Sci Rep. 2020 May 15;10(1):8102. doi: 10.1038/s41598-020-65044-7.

Abstract

Previously, we developed a CHO cell display-based antibody maturation procedure in which an antibody (or other protein) gene of interest was induced to mutate by activation-induced cytidine deaminase (AID) and then form a library by simply proliferating the CHO cells in culture. In this study, we further improved the efficiency of this maturation system by reengineering AID, and optimizing the nucleic acid sequence of the target antibody gene and AID gene as well as the protocol for AID gene transfection. These changes have increased both the mutation rate and the number of mutation type of antibody genes by more than 10 fold, and greatly improved the maturation efficiency of antibody/other proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / immunology
  • CHO Cells
  • Cricetinae
  • Cricetulus
  • Cytidine Deaminase / genetics*
  • Cytidine Deaminase / immunology*
  • Gene Library*
  • Humans
  • Mutation Rate
  • Mutation*
  • Single-Chain Antibodies / biosynthesis*
  • Single-Chain Antibodies / genetics
  • Single-Chain Antibodies / immunology
  • Tumor Necrosis Factor-alpha / antagonists & inhibitors*
  • Tumor Necrosis Factor-alpha / immunology

Substances

  • Antibodies, Monoclonal
  • Single-Chain Antibodies
  • Tumor Necrosis Factor-alpha
  • Cytidine Deaminase