Abstract
AbstractPreviously, 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.
Funder
the Beijing Science and Technology Project under grants
the National Key R&D Program of China under grants
the National Natural Science Foundation of China under grants
Publisher
Springer Science and Business Media LLC
Reference28 articles.
1. Bradbury, A. R. & Marks, J. D. Antibodies from phage antibody libraries. J Immunol Methods. 290, 29–49 (2004).
2. Daugherty, P. S., Bl, O. M. I., Georgiou, G. & Chen, G. Antibody affinity maturation using bacterial surface display. Protein Eng. 11, 825 (1998).
3. Feldhaus, M. & Siegel, R. Flow cytometric screening of yeast surface display libraries. Methods in Molecular Biology. 263, 311–332 (2004).
4. Harvey, B. R. et al. Anchored periplasmic expression, a versatile technology for the isolation of high-affinity antibodies from Escherichia coli-expressed libraries. Proc Natl Acad Sci USA 101, 9193–9198 (2004).
5. Kondo, A. & Ueda, M. Yeast cell-surface display—applications of molecular display. Appl Microbiol Biotechnol. 64, 28–40 (2004).
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