2019 Fiscal Year Final Research Report
NMR method to obtain structural fingerprints of an intact monoclonal antibody acquired under formulated storage conditions
| Project/Area Number |
18K19415
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| Research Category |
Grant-in-Aid for Challenging Research (Exploratory)
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| Allocation Type | Multi-year Fund |
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| Review Section |
Medium-sized Section 47:Pharmaceutical sciences and related fields
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| Research Institution | National Institute of Advanced Industrial Science and Technology |
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Principal Investigator |
Takeuchi Koh 国立研究開発法人産業技術総合研究所, 生命工学領域, 研究グループ長 (20581284)
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| Co-Investigator(Kenkyū-buntansha) |
徳永 裕二 国立研究開発法人産業技術総合研究所, 生命工学領域, 研究員 (80713354)
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| Project Period (FY) |
2018-06-29 – 2020-03-31
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| Keywords | 高分子バイオ医薬 / NMR / 創薬科学 / 高次構造解析 |
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| Outline of Final Research Achievements |
Non-invasive evaluation of tertiary structure is fundamental for the research, development, and use of the biologics. However, few methodologies are currently available for evaluating large molecular weight (MW) biologics, such as ther-apeutic monoclonal antibodies (mAbs; 150 kDa). Here, we have newly developed a 15N direct detection nuclear magnetic resonance (NMR) technique, the 15N direct detection CRINEPT, that allows the observation of the main chain amide reso-nances of a non-deuterated protein with MW 150 kDa. The technique not only substantially expands the range of pro-teins applicable to solution NMR studies, but also allows the non-invasive structural analyses of intact mAbs in a wide range of temperature and solvent conditions. As proof of principle, we successfully acquired the 15N-detected CRINEPT spectra of an intact mAb in its formulated solution at 4°C.
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| Free Research Field |
創薬科学
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| Academic Significance and Societal Importance of the Research Achievements |
本研究では困難であった製剤・保存条件下での高分子バイオ医薬の高次構造評価を、独自のNMR技術：15N観測CRENEPT法の開発により可能にした。その際、代表的な高分子バイオ医薬である抗体について、完全に非破壊的な高次構造解析を低温かつ保存温度というNMR測定に不利な条件で実現することに成功した。本研究の成果は、バイオ医薬の高機能化を含む研究開発への貢献が期待されるが、本手法を用いることでNMR法により解析可能なタンパク質の数を飛躍的に向上させることが可能であるであるため、広く構造生命科学的研究に資するものである。
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