Development of intrabody selection technology in mammalian cells

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K14486
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biofunction/Bioprocess
• Research Institution: The University of Tokyo
• Principal Investigator: Nagamune Teruyuki 東京大学, 大学院工学系研究科(工学部), 教授 (20124373)
• Co-Investigator(Renkei-kenkyūsha): KAWAHARA Masahiro 東京大学, 大学院工学系研究科, 准教授 (50345097)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: バイオテクノロジー / キメラ受容体 / 細胞内抗体スクリーニング / 細胞内蛋白質間相互作用検出 / 細胞アレイ

Outline of Final Research Achievements

In this study, for the development of next-generation antibody drug, we developed a novel versatile intrabody selection method, which can select intrabody against antigen protein expressed in the cytosol of mammalian cells rapidly and directly. We constructed a chimeric receptor library, which can transduce growth signal by binding to antigen, by conjugating naive library of single chain antibody scFv and intracellular domain of receptor c-kit through flexible linker G4S. We expressed this chimeric receptor library together with a model antigen, the nucleoprotein of a rabies virus, in IL-3 dependent mammalian cell Ba/F3, and successfully obtained several scFv binders against the antigen protein from proliferated cells under IL-3 deficient culture condition.

Free Research Field

バイオテクノロジー、細胞工学