湖南师范大学杨荣华团队报道了自催化集成生物正交（聚）催化剂连接的活细胞膜抗原免疫吸附测定。相关研究成果于2024年10月29日发表于国际顶尖学术期刊《德国应用化学》。

免疫测定方法，特别是酶联免疫吸附测定（ELISAs），以其信号放大能力而闻名，广泛应用于科学研究和临床诊断。然而，酶的不稳定性及其对细胞环境的敏感性对ELISA在活细胞中的广泛应用提出了重大挑战。

该文中，研究人员提出了一种用于检测细胞膜抗原的生物正交（多）催化连接免疫吸附试验（BCLISA），该试验将基于小分子或聚合物的生物正交催化剂与抗体偶联。经过筛选，研究人员选择Cu（I）催化叠氮炔烃环加成反应（CuAAC）作为核心反应体系。

由于其多个催化位点，聚合物基催化剂在相同的分子浓度下表现出增强的反应性。CuAAC反应过程中形成的聚三唑具有螯合Cu（I）的能力，从而促进更快的催化作用。

利用这种自催化特性，研究人员成功地提高了BCLISA的信号放大潜力。最终，这种自催化集成的BCLISA技术被用于体外和活细胞膜上的抗原检测和成像。

该方法为活细胞上低丰度抗原的检测和成像提供了一种新方法。

附：英文原文

Title: Autocatalysis-Integrated Bioorthogonal (Poly)Catalyst-Linked Immunosorbent Assay for Living Cell Membrane Antigens

Author: Xuepu Feng, Guoming Tong, Zengwei Ran, Xiaojuan Liu, Liang Li, Guhuan Liu, Ronghua Yang

Issue&Volume: 2024-10-29

Abstract: Immunoassay methods, notably enzyme-linked immunosorbent assays (ELISAs), renowned for their signal amplification capabilities, are extensively employed in scientific research and clinical diagnostics. However, the instability of enzymes and their sensitivity to cellular environments present significant challenges for the broad application of ELISA in living cells. In this work, we present a bioorthogonal (poly)catalysis-linked immunosorbent assay (BCLISA) designed for the detection of cell membrane antigens, which involves coupling bioorthogonal catalysts based on small molecules or polymers to antibodies. After screening, we opted for the copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) as the core reaction system. The polymer-based catalysts exhibit enhanced reactivity at the same molecular concentration due to their multiple catalytic sites. Polytriazoles formed during the CuAAC reaction have the ability to chelate Cu(I), therebypromoting faster catalysis. By harnessing this autocatalytic feature, we successfully increased the signal amplification potential of BCLISA. Ultimately, this autocatalysis-integrated BCLISA technique was employed for antigen detection and imaging on both in vitro and living cell membranes. This approach offers a new method for the detection and imaging of low-abundance antigens on living cells.

DOI: 10.1002/anie.202417352

Source: https://onlinelibrary.wiley.com/doi/10.1002/anie.202417352