Development of antibody-fusion proximity biotinylation enzyme for analysis of virus entry

2022 Fiscal Year Final Research Report

• Project/Area Number: 21K19230
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
• Research Institution: Ehime University
• Principal Investigator: Sawasaki Tatsuya 愛媛大学, プロテオサイエンスセンター, 教授 (50314969)
• Project Period (FY): 2021-07-09 – 2023-03-31
• Keywords: 近位依存性ビオチン標識法 / ウイルス / 感染症 / AirID / ビオチン / 新型コロナウイルス / レンチウイルス

Outline of Final Research Achievements

Using lentivirus as a model, we applied the proximal-dependent biotinyl labeling method to elucidate the virus infection entry pathway, consisting of 1) construction of an anti-virus-AirID fused with a virus recognition antibody Fab, 2) generation of antibody-AirID-loaded virus, 3) verification of biotinylation of virus surface proteins, 4) confirmation of infection ability in cells, and 5) identification of antibody-AirID-loaded We developed technologies for the following five research projects: 1) creation of biotinylated viruses, 2) creation of biotinylated viruses, 3) validation of biotinylation of viral surface proteins, 4) confirmation of virus infectivity, and 5) identification of biotinylated host proteins in cells infected with virus. With this method, the proximal-dependent biotinyl labeling method was applied to elucidate the viral infection entry pathway.

Free Research Field

蛋白質科学

Academic Significance and Societal Importance of the Research Achievements

ウイルスの構造タンパク質を特異的に認識する抗体に近位依存性ビオチン化酵素を融合することにより、ウイルス粒子を近位依存性ビオチン化酵素でラベルし、ウイルスが細胞へ侵入するために関与する宿主タンパク質をビオチン化という形で検出・同定する技術の開発は、世界初の試みであるため挑戦的で社会的意義は大きい。