tetano
Editor, Senior Moderator
Front Microbiol
. 2022 Aug 22;13:968036.
doi: 10.3389/fmicb.2022.968036. eCollection 2022.
Simultaneous measurement of the antibody responses against SARS-CoV-2 and its multiple variants by a phage display mediated immuno-multiplex quantitative PCR-based assay
Hanyi Chen[SUP] 1 [/SUP], Shen Li[SUP] 1 [/SUP], Jiali Wang[SUP] 1 [/SUP], Siqi He[SUP] 1 [/SUP], Dong Wang[SUP] 2 [/SUP], Zhaohui Qian[SUP] 3 [/SUP], Dandan Hu[SUP] 4 [/SUP], Fangfang Qi[SUP] 5 [/SUP], Keping Hu[SUP] 6 7 [/SUP], Chenyi Luo[SUP] 1 8 [/SUP], Jianxun Wang[SUP] 1 8 [/SUP]
Affiliations
Abstract
To combat the continued pandemic of COVID-19, multiplex serological assays have been developed to comprehensively monitor the humoral immune response and help to design new vaccination protocols to different SARS-CoV-2 variants. However, multiplex beads and stably transfected cell lines require stringent production and storage conditions, and assays based on flow cytometry is time-consuming and its application is therefore restricted. Here, we describe a phage display system to distinguish the differences of immune response to antigenic domains of multiple SARS-CoV-2 variants simultaneously. Compared with linear peptides, the recombinant antigens displayed on the phage surface have shown some function that requires the correct folding to form a stable structure, and the binding efficiency between the recombinant phage and existing antibodies is reduced by mutations on antigens known to be important for antigen-antibody interaction. By using Phage display mediated immuno-multiplex quantitative PCR (Pi-mqPCR), the binding efficiency between the antibody and antigens of different SARS-CoV-2 variants can be measured in one amplification reaction. Overall, these data show that this assay is a valuable tool to evaluate the humoral response to the same antigen of different SARS-CoV-2 variants or antigens of different pathogens. Combined with high-throughput DNA sequencing technology, this phage display system can be further applied in monitoring humoral immune response in a large population before and after vaccination.
Keywords: SARS-CoV-2; antibody responses; coronavirus; phage display; variants.
. 2022 Aug 22;13:968036.
doi: 10.3389/fmicb.2022.968036. eCollection 2022.
Simultaneous measurement of the antibody responses against SARS-CoV-2 and its multiple variants by a phage display mediated immuno-multiplex quantitative PCR-based assay
Hanyi Chen[SUP] 1 [/SUP], Shen Li[SUP] 1 [/SUP], Jiali Wang[SUP] 1 [/SUP], Siqi He[SUP] 1 [/SUP], Dong Wang[SUP] 2 [/SUP], Zhaohui Qian[SUP] 3 [/SUP], Dandan Hu[SUP] 4 [/SUP], Fangfang Qi[SUP] 5 [/SUP], Keping Hu[SUP] 6 7 [/SUP], Chenyi Luo[SUP] 1 8 [/SUP], Jianxun Wang[SUP] 1 8 [/SUP]
Affiliations
- PMID: 36071962
- PMCID: PMC9441900
- DOI: 10.3389/fmicb.2022.968036
Abstract
To combat the continued pandemic of COVID-19, multiplex serological assays have been developed to comprehensively monitor the humoral immune response and help to design new vaccination protocols to different SARS-CoV-2 variants. However, multiplex beads and stably transfected cell lines require stringent production and storage conditions, and assays based on flow cytometry is time-consuming and its application is therefore restricted. Here, we describe a phage display system to distinguish the differences of immune response to antigenic domains of multiple SARS-CoV-2 variants simultaneously. Compared with linear peptides, the recombinant antigens displayed on the phage surface have shown some function that requires the correct folding to form a stable structure, and the binding efficiency between the recombinant phage and existing antibodies is reduced by mutations on antigens known to be important for antigen-antibody interaction. By using Phage display mediated immuno-multiplex quantitative PCR (Pi-mqPCR), the binding efficiency between the antibody and antigens of different SARS-CoV-2 variants can be measured in one amplification reaction. Overall, these data show that this assay is a valuable tool to evaluate the humoral response to the same antigen of different SARS-CoV-2 variants or antigens of different pathogens. Combined with high-throughput DNA sequencing technology, this phage display system can be further applied in monitoring humoral immune response in a large population before and after vaccination.
Keywords: SARS-CoV-2; antibody responses; coronavirus; phage display; variants.