Development of houndshark nanodoy by combination of wheat cell-free and NGS technologies

2018 Fiscal Year Final Research Report

• Project/Area Number: 16H04729
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: System genome science
• 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)
• Research Collaborator: Narita Mitsuyoshi 愛媛県栽培資源研究所
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: cell-free / antibody / shark / nanobody / fukabody / houdshark / NGS

Outline of Final Research Achievements

We used GFP (green fluorescent protein) as a model for shark nonobody, which we called shark nanobody as fukabody. After GFP immunization to houndshark, some houndsharks indicated specific immunoresponse to GFP. We screened GFP-binding fukabody by phage display at totally three-time rounds. DNA fragments from each round were analyzed its sequences by next generation sequencing (NGS) technology. Enriched DNA fragments were found, and then their proteins were synthesized by wheat cell-free system. Enriched clones indicated binding ability to GFP antigen. We are successful to develop construction system for fukabody by combination of wheat cell-free and NGS technologies.

Free Research Field

蛋白質科学

Academic Significance and Societal Importance of the Research Achievements

本研究成果により、日本産エイラクブカを用いて、抗原特異的なフカボディを単離する技術が確立できた。フカボディは、ナノボディよりさらに抗原タンパク質の溝を認識することが本研究成果により分かってきたので、フカボディを利用した創薬ができる基盤技術となった。また、本研究成果により古代ザメとは異なり、現在に広く存在するサメは、哺乳類に近い免疫応答系を有していることが明らかとなった。