Construction of efficient systems for generation of anti-metatypic antibody for high-sensitive detection of small molecules
| Project/Area Number |
19K07033
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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) |
森田 いずみ 神戸薬科大学, 薬学部, 講師 (20299085)
小林 典裕 神戸薬科大学, 薬学部, 教授 (90205477)
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| Project Period (FY) |
2019-04-01 – 2022-03-31
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| Project Status |
Completed (Fiscal Year 2021)
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| Budget Amount *help |
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2021: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000)
Fiscal Year 2020: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
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| Keywords | 免疫測定法 / 抗メタタイプ抗体 / scFv / 抗体ライブラリー / ハプテン / サンドイッチELISA / 抗体工学 / ファージ提示 / ファーージ提示 / タンパク質工学 |
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| Outline of Research at the Start |
イムノクロマト法に代表されるサンドイッチ型の免疫測定法は、簡便で多検体の同時処理が可能なため医療機関での迅速な診断に不可欠である。疾患に関連するバイオマーカーの多くは低分子化合物（ハプテン）であるが、その分子サイズゆえにサンドイッチ法への適用は原理的に困難であった。本研究では、ハプテン－抗ハプテン抗体複合体を認識する抗体を創製し、ハプテンの高感度なサンドイッチ法の開発に挑戦する。その際、動物の免疫に頼ることなく、人工の抗体ライブラリーから新規に開発した特異抗体提示ファージの選択法を用いることにより、希少な抗複合体抗体の獲得を可能にする。
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| Outline of Final Research Achievements |
Immunoassay using antigen-antibody reactions are frequently used in clinical testing and diagnostics. Sandwich ELISAs use two different types of antibodies to achieve high sensitivity and specificity, but in principle it has been difficult to apply them to small molecules (haptens). Anti-metatypic antibodies are special antibodies that recognize antigen-antibody complexes and are the key material to establishing sandwich ELISAs for haptens. However, it was difficulted to generation them by the conventional method of immunizing animals. In this study, an array-type screening method was used to prepare an efficient system for generating mutant antibodies that exhibit anti-metatypic activity from antibody-displaying phage libraries. The mutant antibodies obtained enabled sandwich ELISA for thyroxine.
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| Academic Significance and Societal Importance of the Research Achievements |
研究は抗体ライブラリーの活用と目的の抗体分子種を選択する新たな方法の組み合わせであり、類例はほとんどない。免疫複合体に高い選択性を示す抗メタタイプ抗体の効率的な調製は、従来不可能であったステロイドやビタミン類のサンドイッチ型測定法の構築が容易となる。結果として、検体の測定感度が飛躍的に改善されるうえ操作性にも優れ、臨床診断や現場分析に大きく貢献すると思われる。
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Report
(4 results)
Research Products
(25 results)
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[Journal Article] NanoLuc luciferase as a suitable fusion partner of recombinant antibody fragments for developing sensitive luminescent immunoassays2021
Author(s)
Oyama Hiroyuki, Kiguchi Yuki, Morita Izumi, Miyashita Takayuki, Ichimura Akiyoshi, Miyaoka Hiroko, Izumi Aki, Terasawa Sayaka, Osumi Natsumi, Tanaka Hiroki, Niwa Toshifumi, Kobayashi Norihiro
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Journal Title
Analytica Chimica Acta
Volume: 1161
Pages: 238180-238180
DOI
Related Report
Peer Reviewed
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[Journal Article] Clonal array profiling of scFv-displaying phages for high-throughput discovery of affinity-matured antibody mutants2020
Author(s)
Kiguchi Yuki, Oyama Hiroyuki, Morita Izumi, Morikawa Mai, Nakano Asuka, Fujihara Wakana, Inoue Yukari, Sasaki Megumi, Saijo Yuki, Kanemoto Yuki, Murayama Kaho, Baba Yuki, Takeuchi Atsuko, Kobayashi Norihiro
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Journal Title
Scientific Reports
Volume: 10
Issue: 1
Pages: 14103-14103
DOI
Related Report
Peer Reviewed / Open Access
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[Journal Article] Seeking high-priority mutations enabling successful antibody-breeding: systematic analysis of a mutant that gained over 100-fold enhanced affinity2020
Author(s)
Oyama Hiroyuki, Kiguchi Yuki, Morita Izumi, Yamamoto Chika, Higashi Yuka, Taguchi Miku, Tagawa Tatsuya, Enami Yuri, Takamine Yuriko, Hasegawa Hanako, Takeuchi Atsuko, Kobayashi Norihiro
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Journal Title
Scientific Reports
Volume: 10
Issue: 1
Pages: 4807-4807
DOI
Related Report
Peer Reviewed / Open Access
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