| Project/Area Number |
18K07030
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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 49020:Human pathology-related
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| Research Institution | Fujita Health University |
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Principal Investigator |
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| 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)
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| Project Period (FY) |
2018-04-01 – 2022-03-31
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| Project Status |
Completed (Fiscal Year 2021)
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| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2019: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | 酵素抗原法 / 形質細胞 / 抗体 / 抗原 / 疾患特異的抗体 / 抗体産生細胞 / パラフィン切片 / 免疫 / 慢性炎症 / 感染症 / がん / パラフィン包埋切片 / 固定 |
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| 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.
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| 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。
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