Bernauer Hubert, Maier Josef, Bannert Norbert, Ivanusic Daniel
ATG:biosynthetics GmbH, 79249 Merzhausen, Germany.
Sexually Transmitted Bacterial Pathogens and HIV (FG18), Robert Koch-Institute, 13353 Berlin, Germany.
Biol Methods Protoc. 2024 Jan 19;9(1):bpae001. doi: 10.1093/biomethods/bpae001. eCollection 2024.
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 severe acute respiratory syndrome coronavirus 2 (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 are not necessary. With this technique, the immune evasion of new SARS-CoV-2 variants from current vaccination regimes can be examined quickly and reliably.
酶联免疫吸附测定(ELISA)系统使用包被有肽或表达并纯化的蛋白质的平板来监测源自患者血清的免疫球蛋白。然而,目前尚无一种简单、灵活且快速的基于ELISA的适应性系统来检测针对新型严重急性呼吸综合征冠状病毒2(SARS-CoV-2)变体的抗体。在本研究中,我们利用了tANCHOR蛋白展示系统,该系统提供了一个装饰有受体结合域(RBD)的细胞表面,以监测来自SARS-CoV-2康复者和接种疫苗个体的针对它的特异性抗体。为了检测来自疫苗接种者或康复个体的血清,只需将RBD编码序列克隆到tANCHOR载体系统中并转染到HeLa细胞中。无需进行耗时的蛋白质表达、分离和纯化,然后再包被测定平板。通过这项技术,可以快速可靠地检测新的SARS-CoV-2变体对当前疫苗接种方案的免疫逃逸情况。