Efficient systems for generating high-performance antibody mutants that enable sensitive determinations of diagnostic biomarkers
| Project/Area Number |
15K08629
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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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| Research Field |
Laboratory medicine
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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, 助教 (80572966)
森田 いずみ 神戸薬科大学, 薬学部, 助手 (20299085)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2017: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2016: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 抗体 / 一本鎖Fvフラグメント / 変異体 / ライブラリー / 選択方法 / パンニング / 変異抗体 / バイオマーカー / 進化分子工学 / ファージ提示 / マイクロアレイ / デジタル検出 |
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| Outline of Final Research Achievements |
Phage-display-based molecular evolution potentially provides antibody mutants with greater diagnostic utilities than conventional antibodies. The panning, in which whole members were reacted simultaneously to immobilized antigens, has still been used to isolate evolved mutants in diverse antibody libraries, but often missed valuable species. Thus, we developed a new selection strategy to solve this crucial problem. Bacterial clones in a library, each harboring a single-chain Fv fragment (scFv) gene, were cultured separately in microwells, and scFv-displaying phage generated therein was examined for their antigen-binding property using a bioluminescent assay. One-shot application for an scFv library against cortisol provided several scFv clones with improved affinities, none of which could be isolated through a repeated trials of conventional panning. Thus we evaluated that the present method should be of significant utility for generating high-performance diagnostic antibodies.
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Report
(4 results)
Research Products
(33 results)
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[Journal Article] One-Shot In Vitro Evolution Generated an Antibody Fragment for Testing Urinary Cotinine with More Than 40-Fold Enhanced Affinity2017
Author(s)
Oyama, Hiroyuki; Morita, Izumi; Kiguchi, Yuki; Banzono, Erika; Ishii, Kasumi; Kubo, Satoshi; Watanabe, Yoshiro; Hirai, Anna; Kaede, Chiaki; Ohta, Mitsuhiro; Kobayashi Norihiro
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Journal Title
Analytical Chemistry
Volume: 89(1)
Issue: 1
Pages: 988-995
DOI
Related Report
Peer Reviewed
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[Journal Article] Immunochemical Approach for Monitoring of Structural Transition of ApoA-I upon HDL Formation Using Novel Monoclonal Antibodies2017
Author(s)
H. Kimura, S. Mikawa, C. Mizuguchi, Y. Horie, I. Morita, H. Oyama, T. Ohgita, K. Nishitsuji, A. Takeuchi, S. Lund-Katz, K. Akaji, N. Kobayashi, H. Saito
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Journal Title
Sci. Rep.
Volume: 7
Issue: 1
Pages: 2988-2988
DOI
NAID
Related Report
Peer Reviewed / Open Access / Int'l Joint Research
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[Journal Article] The polyphenol (-)-epigallocatechin-3-gallate prevents apoA-IIowa amyloidosis in vitro and protects human embryonic kidney 293 cells against amyloid cytotoxicity.2016
Author(s)
Nakajima H, Nishitsuji K, Kawashima H, Kuwabara K, Mikawa S, Uchimura K, Akaji K, Kashiwada Y, Kobayashi N, Saito H, Sakashita N
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Journal Title
Amyloid
Volume: 23
Issue: 1
Pages: 17-25
DOI
Related Report
Peer Reviewed
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[Journal Article] Two chalcones, 4-hydroxyderricin and xanthoangelol, stimulate GLUT4-dependent glucose uptake through the LKB1/AMP-activated protein kinase signaling pathway in 3T3-L1 adipocytes2015
Author(s)
Ohta, Mitsuhiro; Fujinami, Aya; Kobayashi, Norihiro; Amano, Akiko; Ishigami, Akihito; Tokuda, Harukuni; Suzuki, Nobutaka; Ito, Fumitake; Mori, Taisuke; Sawada, Morio;
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Journal Title
Nutrition Research
Volume: 35(7)
Issue: 7
Pages: 618-625
DOI
Related Report
Peer Reviewed
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[Journal Article] Cellular Interaction and Cytotoxicity of the Iowa Mutation of Apolipoprotein A-I (ApoA-IIowa) Amyloid Mediated by Sulfate Moieties of Heparan Sulfate2015
Author(s)
Kuwabara, K., Nishitsuji, K., Uchimura, K., Hung, S.C., Mizuguchi, M., Nakajima, H., Mikawa, S., Kobayashi, N., Saito, H., and Sakashita, N.
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Journal Title
J Biol Chem
Volume: 290
Issue: 40
Pages: 24210-24221
DOI
Related Report
Peer Reviewed / Int'l Joint Research
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[Presentation] Effect of Heparin on Amyloid Fibril Formation of ApoA-I Fragment Peptides2016
Author(s)
Shiho Mikawa, Chiharu Mizuguchi, Izumi Morita, Hioryuki Oyama, Teruhiko Baba, Akira Shigenaga, Toshinori Shimanouchi, Norihiro Kobayashi, Akira Otaka, Kenichi Akaji, Hiroyuki Saito
Organizer
第53回ペプチド討論会
Place of Presentation
京都
Year and Date
2016-10-26
Related Report
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