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

IF 16.1 1区 化学 Q1 CHEMISTRY, MULTIDISCIPLINARY
Xuepu Feng, Guoming Tong, Zengwei Ran, Xiaojuan Liu, Liang Li, Guhuan Liu, Ronghua Yang
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引用次数: 0

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.

用于活细胞膜抗原的自催化-集成生物正交(多)催化剂联结免疫吸附试验。
免疫测定方法,特别是酶联免疫吸附测定法(ELISA),以其信号放大能力而闻名,被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给 ELISA 在活细胞中的广泛应用带来了巨大挑战。在这项工作中,我们提出了一种生物正交(多)催化连接免疫吸附试验(BCLISA),该试验设计用于检测细胞膜抗原,包括将基于小分子或聚合物的生物正交催化剂与抗体耦合。经过筛选,我们选择了铜(I)催化的叠氮-炔环加成(CuAAC)作为核心反应体系。聚合物基催化剂由于具有多个催化位点,因此在相同分子浓度下具有更高的反应活性。在 CuAAC 反应过程中形成的聚三唑具有螯合 Cu(I)的能力,从而加快了催化速度。通过利用这种自催化特性,我们成功地提高了 BCLISA 的信号放大潜力。最终,这种自催化集成 BCLISA 技术被用于体外和活细胞膜上的抗原检测和成像。这种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。
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来源期刊
CiteScore
26.60
自引率
6.60%
发文量
3549
审稿时长
1.5 months
期刊介绍: Angewandte Chemie, a journal of the German Chemical Society (GDCh), maintains a leading position among scholarly journals in general chemistry with an impressive Impact Factor of 16.6 (2022 Journal Citation Reports, Clarivate, 2023). Published weekly in a reader-friendly format, it features new articles almost every day. Established in 1887, Angewandte Chemie is a prominent chemistry journal, offering a dynamic blend of Review-type articles, Highlights, Communications, and Research Articles on a weekly basis, making it unique in the field.
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