| 研究課題/領域番号 |
21K14468
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| 研究種目 |
若手研究
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| 配分区分 | 基金 |
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| 審査区分 |
小区分27040:バイオ機能応用およびバイオプロセス工学関連
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| 研究機関 | 東京工業大学 |
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研究代表者 |
朱 博 東京工業大学, 科学技術創成研究院, 助教 (70886605)
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| 研究期間 (年度) |
2021-04-01 – 2024-03-31
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| 研究課題ステータス |
交付 (2022年度)
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| 配分額 *注記 |
3,510千円 (直接経費: 2,700千円、間接経費: 810千円)
2022年度: 1,950千円 (直接経費: 1,500千円、間接経費: 450千円)
2021年度: 1,560千円 (直接経費: 1,200千円、間接経費: 360千円)
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| キーワード | Immunosensor / mRNA display / Quenchbody / SARS-CoV-2 / Phage display / NGS / バイオテクノロジー / タンパク質 / バイオセンサー / 蛍光 / mRNA-display |
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| 研究開始時の研究の概要 |
Quenchbody（Q-body）は, 抗体あるいはその断片を部位特異的に蛍光修飾して得られる, 蛍光免疫センサーである. Q-bodyは, クエンチ解消原理に基づき様々な抗原を迅速・簡便に検出可能と考えられている. しかし, 現状では, 一つの抗原が一つのQ-bodyの蛍光色素を光らせる事しかできず, Q-bodyの感度は, 親抗体の動力学的特性によって制限されると考えられる. 本研究では, Q-bodyの感度を短期間で向上させるためのツールキットを開発することを目的とする. このツールキットにより, Q-bodyに基づく免疫測定法の開発期間を短縮し, 緊急事態への迅速な対応を可能にする.
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| 研究実績の概要 |
The research project is about making a toolkit for improving the performance of Quenchbody-based immunosensor (Quenchbody) in a short period. It will increase the feasibility of the quenchbody while responding to the needs of society in a fast manner, such as combating pandemics.
In the second year, the mRNA-display procedures were extensively optimized to get a functional and large Quenchbody for the linker screening. The cell-free synthesized Bone Gla protein (BGP) antibody fragment was successfully labeled with a fluorophore in an mRNA-display-required working procedure. The affinity and immunosensor function of the labeled antibody were confirmed. In addition, a phage-display-based Quenchbody library was created with unexpectedly good feasibility for functional linker sequence enrichment. Two rounds of selection of a BGP Quenchbody library with the randomized fluorophore linker sequences were performed and a clear motif was enriched, which was confirmed via next-generation sequencing.
In the meanwhile, the crowding agent function for rapidly improving the Quenchbody function was fully tested on a SARS-CoV-2 Quenchbody. The related results have been published in an International journal and the paper was selected as HOT Article and a Back Cover.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
3: やや遅れている
理由
The research progress was slightly affected by the COVID-19 pandemic. An alternative phage display-based Quenchbody library was successfully created with unexpectedly better feasibility to screen linker sequences. Therefore, additional experiment is needed to finalize the rest of the screening and analysis for a higher grade outcome.
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| 今後の研究の推進方策 |
The mRNA-display procedures for Quenchbody will be further optimized for producing a useful Quenchbody library. The alternative and robust Quenchbody-displayed phage library will be screened to find better linker sequences for improved response and signal amplification. One of the targeting antibodies will be shifted from the cortisol antibody to another biomarker thyroxine with better affinity. Instead of the linear model, a machine learning-based model will be generated to predict the function of the linkers by using the next-generation sequence datasets. The related output will be organized as another research article for publication and conference presentation.
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