Applicaton of enzyme-labled antigen method for formalin-fixed paraffin-embedded tissue section

2021 Fiscal Year Final Research Report

• Project/Area Number: 18K07030
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 49020:Human pathology-related
• Research Institution: Fujita Health University
• Principal Investigator: Mizutani Yasuyoshi 藤田医科大学, 医学部, 助教 (10546229)
• Co-Investigator(Kenkyū-buntansha): 塩竈 和也 藤田医科大学, 医療科学部, 講師 (10387699) ; 稲田 健一 藤田医科大学, 医学部, 教授 (70246081) ; 鈴木 元 藤田医科大学, 医学部, 教授 (80236017)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Keywords: 酵素抗原法 / 形質細胞 / 抗体 / 抗原

Outline of Final Research Achievements

In this study, we attempted to apply the enzyme-labeled antigen method to formalin-fixed paraffin-embedded (FFPE) tissue sections. We evaluated retrieval methods for the activity of antibodies on FFPE sections. High-concentration proteinase K treatment resulted in only around 50% retrieval of activity in FFPE sections compared to PFA-fixed frozen sections, but around 80% in PFA-fixed paraffin-embedded sections. We also evaluated whether various fixation methods can maintain antibody activity in paraffin sections. It was found that some fixation methods could maintain antibody activity on paraffin sections regardless of the type of target antigens. Signal amplification methods were also tested and their effectiveness was confirmed.

Free Research Field

病理学

Academic Significance and Societal Importance of the Research Achievements

複数種の固定法について検討を行い、パラフィン切片で抗体活性を維持可能な固定法が明らかとなった。これは、今後の検体の保管管理の簡便化につながる知見であり、酵素抗原法の応用性を高める知見である。また、FFPE切片における抗原の賦活化法や抗原抗体反応のシグナル増感法、あるいは抗原の標識分子に関する有用性を明らかにすることができた。これらの手法を改良して組み合わせることで、酵素抗原法のFFPE切片への応用が可能となることが期待される。