Generation of "antibody-like" proteins based on evolutionary molecular engineering for novel clinical diagnostics

2015 Fiscal Year Final Research Report

• Project/Area Number: 25460709
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Laboratory medicine
• Research Institution: Kobe Pharmaceutical University
• Principal Investigator: MORITA IZUMI 神戸薬科大学, 薬学部, 助手 (20299085)
• Co-Investigator(Kenkyū-buntansha): KOBAYASHI Norihiro 神戸薬科大学, 薬学部, 教授 (90205477) ; OYAMA Hiroyuki 神戸薬科大学, 薬学部, 助教 (80572966)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: 進化分子工学

Outline of Final Research Achievements

Antibodies are widely used in various clinical tests. However, lower expression levels in E. coli and their nature requiring two variable domains (VH and VL) for building the antigen-binding sites reduce their utility in genetic manipulation for generating improved species. Therefore, we attempted creation of new "antibody-like" proteins, which are composed of a single domain scaffold with much smaller molecular weight and can be expressed with a satisfactory yield in E. coli. We chose the biotin-binding protein, streptavidin monomer (stav'), as the basic scaffold. After several steps of mutagenesis and selection for their loop structures protruding from the scaffold, we succeeded in obtaining stav' mutants that had gained novel binding specificities targeting a steroid, estradiol.

Free Research Field

分析化学