tetano
Editor, Senior Moderator
Biol Methods Protoc
. 2024 Jan 19;9(1):bpae001.
doi: 10.1093/biomethods/bpae001. eCollection 2024. 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
Hubert Bernauer[SUP] 1 [/SUP], Josef Maier[SUP] 1 [/SUP], Norbert Bannert[SUP] 2 [/SUP], Daniel Ivanusic[SUP] 2 [/SUP]
Affiliations
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.
Keywords: ELISA; IgG antibodies; SARS-CoV-2; receptor binding domain (RBD); tANCHOR; tetraspanin.
. 2024 Jan 19;9(1):bpae001.
doi: 10.1093/biomethods/bpae001. eCollection 2024. 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
Hubert Bernauer[SUP] 1 [/SUP], Josef Maier[SUP] 1 [/SUP], Norbert Bannert[SUP] 2 [/SUP], Daniel Ivanusic[SUP] 2 [/SUP]
Affiliations
- PMID: 38332985
- PMCID: PMC10850845
- DOI: 10.1093/biomethods/bpae001
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.
Keywords: ELISA; IgG antibodies; SARS-CoV-2; receptor binding domain (RBD); tANCHOR; tetraspanin.