Development of new technology identifying cancer mutation to clarify molecular pathology of cancer

• Project/Area Number: 15H06917
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: Tumor diagnostics
• Research Institution: National Institutes of Biomedical Innovation, Health and Nutrition
• Principal Investigator: Mori Takahide 国立研究開発法人医薬基盤・健康・栄養研究所, 医薬基盤研究所 創薬デザイン研究センター, 技術補助員 (40756847)
• Project Period (FY): 2015-08-28 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: がん / 自己抗体 / 病態解明

Outline of Final Research Achievements

I successfully established a technique recovering a pair of variable heavy and light antibody gene sequences from each B cell in human blood. This technique should be useful to identify autoantibody sequences against mutated cancer antigens. We showed that 2 days-polyclonal activation of B cell from human blood in cell culture followed by single cell sorting dramatically increased the rate of antibody gene recovery from a single cell in the next cDNA preparation and sequencing step. The high rate of recovery of antibody sequence makes this technique practical for obtaining antibody sequence from a limited number of B cell available in patients’ specimens and having their moderate-level of the specific binding affinity. This approach should be practical and applicable to obtain a small size of antibody library from a few patients’ specific B cells in human blood. Isolated antibodies by this method will be useful for identifying autoantigens including mutated cancer-related antigens.

Research Products

• [Presentation] がん分子病態の解明を目指したがん患者由来抗体ライブラリの構築と評価 (2016)
  • Author(s): 芳賀 優弥, 長野 一也, 森 宣瑛, 永野 貴士, 向井 美穂, 大須賀 絵理, 竹谷 苑子, 井阪 亮, 鎌田 春彦, 角田 慎一, 堤 康央
  • Organizer: 日本薬学会第136年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2016-03-26