New screening method for fish monoclonal antibody gene and establishment of antibody expression system

2017 Fiscal Year Final Research Report

• Project/Area Number: 15H02461
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Aquatic life science
• Research Institution: The University of Tokyo
• Principal Investigator: Asakawa Shuichi 東京大学, 大学院農学生命科学研究科(農学部), 教授 (30231872)
• Co-Investigator(Kenkyū-buntansha): 大政 健史 大阪大学, 工学(系)研究科(研究院), 准教授 (00252586) ; 近藤 秀裕 東京海洋大学, 学術研究院, 准教授 (20314635) ; 田角 聡志 東京大学, 大学院農学生命科学研究科(農学部), 特任助教 (90359646)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: IgNAR / 一本鎖抗体 / サメ抗体 / 免疫クロブリン / トランスクリプトーム / CHO細胞 / VLR / トラフグ

Outline of Final Research Achievements

The cDNA sequence of IgNAR of bamboo shark (Chiloscyllium punctatum) was obtained and the sequence of the constant region was extracted. We also established a system to amplify the variable region of IgNAR and measure the expression frequency of individual genes.
Antigens (HEL, BSA, MMP-09) were administered several times to the bamboo shark at intervals of one month, and blood was collected one month after administration. Antibody titer was measured by ELISA for BSA and multiple genes with increased expression frequency were identified in conjunction with rise in antibody titer. Expression constructs combining variable and constant regions of shark antibodies were prepared using CHO cells and succeeded in secretory production. From these results, identification of the antigen-specific antibody gene by transcriptome and establishment of its expression system were almost completed.

Free Research Field

水圏生物工学