2021 Fiscal Year Final Research Report
Construction of innovative strategy for selecting improved antibody fragments as the platform generating next-generation diagnostic reagents
| Project/Area Number |
19K07021
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Review Section |
Basic Section 47020:Pharmaceutical analytical chemistry and physicochemistry-related
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| Research Institution | Kobe Pharmaceutical University |
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Principal Investigator |
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| Co-Investigator(Kenkyū-buntansha) |
大山 浩之 神戸薬科大学, 薬学部, 准教授 (80572966)
森田 いずみ 神戸薬科大学, 薬学部, 講師 (20299085)
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| Project Period (FY) |
2019-04-01 – 2022-03-31
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| Keywords | 診断薬 / 抗体 / 一本鎖Fvフラグメント / 遺伝子工学 / ファージ提示 |
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| Outline of Final Research Achievements |
Much attention has been paid to antibody mutants generated via genetic engineering, which are selected and isolated from “antibody libraries” composed of diverse molecules with a variety of mutations. In this study, we succeeded in developing a novel “clonal array profiling (CAP)” method that dramatically facilitates generation of antibody mutants with improved antigen-binding characteristics. We demonstrated the utility of the CAP method with the following successful applications: (1) we obtained scFv (single-chain Fv fragment) mutants specific to cortisol with 17-31-fold increased affinity constant (Ka) from libraries composed of scFv members with site-directed mutations or amino-acid insertion in their framework region 1 in the VH domain, and (2) we discovered a scFv mutant specific to estradiol with 372-fold enhanced Ka from a library of scFv members with site-directed mutations in the complementarity-determining region 3 in the VH domain.
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| Free Research Field |
分析化学
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| Academic Significance and Societal Importance of the Research Achievements |
遺伝子操作による改変抗体の有用性に期待が寄せられて久しい。しかし、現状では診断用抗体は今なおハイブリドーマ法により生産され、改変抗体はほとんど市場に出回っていない。研究代表者が開発したCAP法は、抗体ライブラリーに含まれる希少な改良型変異体の選択を飛躍的に効率化する。本研究は、改良型変異体の選択法として標準とされてきた「パンニング」の限界を示した点でも意義が大きい。CAP法は遺伝子操作による機能性分子創製の潜在力を開花させ、ハイブリドーマ抗体を超える優れた改変抗体の創製と実用化を促すだろう。ひいては、診断薬のみならず抗体医薬やワクチンの開発も活性化され、医療の進歩にも貢献するものと期待される。
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