Development of luminescence probes for specific detection of human proteins

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K15421
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 37030:Chemical biology-related
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: Nishihara Ryo 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 (50846988)
• Project Period (FY): 2020-04-01 – 2022-03-31
• Keywords: 生物発光 / ルシフェリン / ルシフェラーゼ / タンパク質 / 酵素反応 / 化学センサー / 発光

Outline of Final Research Achievements

We report the design and synthesis of an imidazopyrazinone-type luciferin, Human Luminophore1 (HuLumino1), with the aim of unmasking the latent luciferase activity of non-luminous proteins. The investigation of reaction behavior of HuLumino1 for the biological proteins revealed that only human serum albumin (HSA) led to a distinct luminescent enhancement, while other proteins resulted in no emission. HSA levels were quantified within 5% error margins of an enzyme-linked immunosorbent assay without the need for any sample pretreatments because of the high specificity of HuLumino1. These results indicate the advantages of the novel platform for protein analysis to detect non-labeled proteins, which generally do not function as enzymes. In addition, the "luciferase-independent" luminescence assay can also be used to detect other biomarkers, such as copper ions.

Free Research Field

分析化学

Academic Significance and Societal Importance of the Research Achievements

非発光タンパク質と発光反応を起こす人工型ルシフェリンを開発する独自の研究アプローチで、ルシフェリンを混合するだけで、特異的かつ迅速に、更には従来法(ELISA)と同等の感度で目的タンパク質または低分子バイオマーカーを発光検出できる新規分析技術の基礎基盤を確立する事に成功した。本技術は、従来のタンパク質分析技術とは原理的にも全く異なるもので、今後のタンパク質分析技術の発展に大きく貢献するものだと考える。