tANCHOR cell-based ELISA approach as a surrogate for antigen-coated plates to monitor specific IgG directed to the SARS-CoV-2 receptor binding domain

IF 2.5 Q3 BIOCHEMICAL RESEARCH METHODS
Hubert Bernauer, Josef Maier, Norbert Bannert, Daniel Ivanusic
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引用次数: 0

Abstract

Enzyme-linked immunosorbent assay (ELISA) systems use plates coated with peptides or expressed and purified proteins to monitor immunoglobulins derived from patient serum. However, there is currently no easy, flexible, and fast adaptive ELISA-based system for testing antibodies directed against new SARS-CoV-2 variants. In this study, we utilized the tANCHOR protein display system that provides a cell surface decorated with the receptor binding domain (RBD) to monitor specific antibodies derived from SARS-CoV-2 convalescent and vaccinated individuals directed against it. To test sera from vaccinees or convalescent individuals, only the RBD coding sequence needs to be cloned in the tANCHOR vector system and transfected into HeLa cells. Time-consuming protein expression, isolation, and purification followed by coating assay plates is not necessary. With this technique, the immune evasion of new SARS-CoV-2 variants from current vaccination regimes can be examined quickly and reliably.
用基于 tANCHOR 细胞的 ELISA 方法替代抗原涂层板,监测针对 SARS-CoV-2 受体结合域的特异性 IgG
酶联免疫吸附试验(ELISA)系统使用涂有肽或表达和纯化蛋白质的平板来监测从病人血清中提取的免疫球蛋白。然而,目前还没有一种简便、灵活、快速的自适应 ELISA 系统来检测针对 SARS-CoV-2 新变种的抗体。在这项研究中,我们利用了 tANCHOR 蛋白显示系统,该系统提供了一个由受体结合域(RBD)装饰的细胞表面,用于监测来自 SARS-CoV-2 康复者和疫苗接种者的针对 SARS-CoV-2 的特异性抗体。要检测疫苗接种者或康复者的血清,只需在 tANCHOR 载体系统中克隆 RBD 编码序列并转染 HeLa 细胞。无需进行耗时的蛋白质表达、分离和纯化,然后再涂布检测板。有了这项技术,就可以快速可靠地检测当前疫苗接种方案中新的 SARS-CoV-2 变体的免疫逃避能力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Biology Methods and Protocols
Biology Methods and Protocols Agricultural and Biological Sciences-Agricultural and Biological Sciences (all)
CiteScore
3.80
自引率
2.80%
发文量
28
审稿时长
19 weeks
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