A rapid screening and production method using a novel mammalian cell display to isolate human monoclonal antibodies

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• 作者：Kosuke Tomimatsu ; Shin-ei Matsumoto ; Hayato Tanaka ; Makiko Yamashita ; Hidekazu Nakanishi ; Kiichiro Teruya ; Saiko Kazuno ; Tomoya Kinjo ; Takeki Hamasaki ; Ken-ichi Kusumoto ; Shigeru Kabayama ; Yoshinori Katakura ; Sanetaka Shirahata
• 关键词：Human monoclonal antibody ; Mammalian cell display ; In vitro maturation ; CHO cells ; Cre recombinase ; Human high affinity IgE receptor
• 刊名：Biochemical and Biophysical Research Communications
• 出版年：8 November, 2013
• 年：2013
• 卷：441
• 期：1
• 页码：59-64
• 全文大小：1058 K

文摘

Antibody display methods are increasingly being used to produce human monoclonal antibodies for disease therapy. Rapid screening and isolation of specific human antibody genes are valuable for producing human monoclonal antibodies showing high specificity and affinity. In this report, we describe a novel mammalian cell display method in which whole human IgG is displayed on the cell surface of CHO cells. Cells expressing antigen-specific human monoclonal IgGs with high affinity on the cell surface after normal folding and posttranscriptional modification were screened using a cell sorter. The membrane-type IgG-expressing CHO cells were then converted to IgG-secreting cells by transfection with a plasmid coding Cre recombinase. This mammalian cell display method was applied to in vitro affinity maturation of monoclonal C9 IgG specific to the human high-affinity IgE receptor (Fc蔚RI伪). The CDR3 of the C9 heavy chain variable region gene was randomly mutated and inserted into pcDNA5FRT/IgG. A C9 IgG (CDRH3r)-expressing CHO cell display library consisting of 1.1 脳 10⁶ independent clones was constructed. IgG-displaying cells showing high reactivity to Fc蔚RI伪 antigen were screened by the cell sorter, resulting in the establishment of a CHO cell line producing with higher reactivity than the parent C9 IgG.