Establishment of a new technique for immunocytochemistry: microparticle-labeled antibody staining

2023 Fiscal Year Final Research Report

• Project/Area Number: 22K18227
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 90140:Medical technology assessment-related
• Research Institution: Nagoya University
• Principal Investigator: Sakabe Nanako 名古屋大学, 医学系研究科(保健), 助教 (90907809)
• Project Period (FY): 2022-04-01 – 2024-03-31
• Keywords: 微粒子標識抗体染色 / 免疫組織化学 / 膜抗原 / 非特異反応 / ワンステップ法 / ツーステップ法

Outline of Final Research Achievements

The purpose of this study is to establish a simple and general-purpose, microparticle-labeled antibody staining, that does not require a staining step. We produced a secondary antibody labeled with microparticles and performed immunostaining (two-step method) using cultured cell lines, resulting in the detection of the target antigen without the staining step. In this study, we detected several membrane antigens using the microparticle-labeled antibody. We suggest that the use of microparticles of different colors may lead to multiplex staining to detect different antigens on the same specimen. In addition, we evaluated the application of the two-step method to rapid immunostaining and obtained positive staining results in only 10 minutes of antibody reactions. We consider that microparticle-labeled antibody staining might be applicable to intraoperative diagnosis.

Free Research Field

医療技術評価学

Academic Significance and Societal Importance of the Research Achievements

医療分野における診断・治療薬適応判定や研究分野で広く使用される免疫染色は、操作が煩雑で時間を要し、非特異反応や発色操作による過染色・未染色などの問題も存在する。本研究において、様々な色の微粒子を用いて微粒子標識抗体を作製し、培養細胞株を用いた検討により目的の抗原を発色操作なしで検出することができた。これにより、染色作業が簡便・迅速化され過染色・未染色による誤判定も回避でき、術中細胞診断への応用にも繋がると考えている。さらに、微粒子の色を使い分けることで同一標本上で複数の抗原を観察する多重染色へも応用でき、汎用性の高い染色法の提供に繋がると考えている。