tetano
Editor, Senior Moderator
Clin Transl Immunology
. 2022 Mar 23;11(3):e1380.
doi: 10.1002/cti2.1380. eCollection 2022.
A scalable serology solution for profiling humoral immune responses to SARS-CoV-2 infection and vaccination
Karen Colwill[SUP] 1 [/SUP], Yannick Galipeau[SUP] 2 [/SUP], Matthew Stuible[SUP] 3 [/SUP], Christian Gervais[SUP] 3 [/SUP], Corey Arnold[SUP] 2 [/SUP], Bhavisha Rathod[SUP] 1 4 [/SUP], Kento T Abe[SUP] 1 5 [/SUP], Jenny H Wang[SUP] 1 [/SUP], Adrian Pasculescu[SUP] 1 [/SUP], Mariam Maltseva[SUP] 2 [/SUP], Lynda Rocheleau[SUP] 2 [/SUP], Martin Pelchat[SUP] 2 6 [/SUP], Mahya Fazel-Zarandi[SUP] 1 [/SUP], Mariam Iskilova[SUP] 1 [/SUP], Miriam Barrios-Rodiles[SUP] 1 [/SUP], Linda Bennett[SUP] 1 [/SUP], Kevin Yau[SUP] 7 [/SUP], François Cholette[SUP] 8 9 [/SUP], Christine Mesa[SUP] 8 [/SUP], Angel X Li[SUP] 1 10 [/SUP], Aimee Paterson[SUP] 1 10 [/SUP], Michelle A Hladunewich[SUP] 7 [/SUP], Pamela J Goodwin[SUP] 1 11 [/SUP], Jeffrey L Wrana[SUP] 1 5 [/SUP], Steven J Drews[SUP] 12 13 [/SUP], Samira Mubareka[SUP] 14 15 16 17 [/SUP], Allison J McGeer[SUP] 1 10 18 [/SUP], John Kim[SUP] 8 [/SUP], Marc-André Langlois[SUP] 2 6 [/SUP], Anne-Claude Gingras[SUP] 1 5 [/SUP], Yves Durocher[SUP] 3 [/SUP]
Affiliations
Abstract
Objectives: Antibody testing against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been instrumental in detecting previous exposures and analyzing vaccine-elicited immune responses. Here, we describe a scalable solution to detect and quantify SARS-CoV-2 antibodies, discriminate between natural infection- and vaccination-induced responses, and assess antibody-mediated inhibition of the spike-angiotensin converting enzyme 2 (ACE2) interaction.
Methods: We developed methods and reagents to detect SARS-CoV-2 antibodies by enzyme-linked immunosorbent assay (ELISA). The main assays focus on the parallel detection of immunoglobulin (Ig)Gs against the spike trimer, its receptor binding domain (RBD) and nucleocapsid (N). We automated a surrogate neutralisation (sn)ELISA that measures inhibition of ACE2-spike or -RBD interactions by antibodies. The assays were calibrated to a World Health Organization reference standard.
Results: Our single-point IgG-based ELISAs accurately distinguished non-infected and infected individuals. For seroprevalence assessment (in a non-vaccinated cohort), classifying a sample as positive if antibodies were detected for ≥ 2 of the 3 antigens provided the highest specificity. In vaccinated cohorts, increases in anti-spike and -RBD (but not -N) antibodies are observed. We present detailed protocols for serum/plasma or dried blood spots analysis performed manually and on automated platforms. The snELISA can be performed automatically at single points, increasing its scalability.
Conclusions: Measuring antibodies to three viral antigens and identify neutralising antibodies capable of disrupting spike-ACE2 interactions in high-throughput enables large-scale analyses of humoral immune responses to SARS-CoV-2 infection and vaccination. The reagents are available to enable scaling up of standardised serological assays, permitting inter-laboratory data comparison and aggregation.
Keywords: SARS‐CoV‐2; antibody detection; antibody neutralisation; assay development and standardisation; high‐throughput screening.
. 2022 Mar 23;11(3):e1380.
doi: 10.1002/cti2.1380. eCollection 2022.
A scalable serology solution for profiling humoral immune responses to SARS-CoV-2 infection and vaccination
Karen Colwill[SUP] 1 [/SUP], Yannick Galipeau[SUP] 2 [/SUP], Matthew Stuible[SUP] 3 [/SUP], Christian Gervais[SUP] 3 [/SUP], Corey Arnold[SUP] 2 [/SUP], Bhavisha Rathod[SUP] 1 4 [/SUP], Kento T Abe[SUP] 1 5 [/SUP], Jenny H Wang[SUP] 1 [/SUP], Adrian Pasculescu[SUP] 1 [/SUP], Mariam Maltseva[SUP] 2 [/SUP], Lynda Rocheleau[SUP] 2 [/SUP], Martin Pelchat[SUP] 2 6 [/SUP], Mahya Fazel-Zarandi[SUP] 1 [/SUP], Mariam Iskilova[SUP] 1 [/SUP], Miriam Barrios-Rodiles[SUP] 1 [/SUP], Linda Bennett[SUP] 1 [/SUP], Kevin Yau[SUP] 7 [/SUP], François Cholette[SUP] 8 9 [/SUP], Christine Mesa[SUP] 8 [/SUP], Angel X Li[SUP] 1 10 [/SUP], Aimee Paterson[SUP] 1 10 [/SUP], Michelle A Hladunewich[SUP] 7 [/SUP], Pamela J Goodwin[SUP] 1 11 [/SUP], Jeffrey L Wrana[SUP] 1 5 [/SUP], Steven J Drews[SUP] 12 13 [/SUP], Samira Mubareka[SUP] 14 15 16 17 [/SUP], Allison J McGeer[SUP] 1 10 18 [/SUP], John Kim[SUP] 8 [/SUP], Marc-André Langlois[SUP] 2 6 [/SUP], Anne-Claude Gingras[SUP] 1 5 [/SUP], Yves Durocher[SUP] 3 [/SUP]
Affiliations
- PMID: 35356067
- PMCID: PMC8942165
- DOI: 10.1002/cti2.1380
Abstract
Objectives: Antibody testing against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been instrumental in detecting previous exposures and analyzing vaccine-elicited immune responses. Here, we describe a scalable solution to detect and quantify SARS-CoV-2 antibodies, discriminate between natural infection- and vaccination-induced responses, and assess antibody-mediated inhibition of the spike-angiotensin converting enzyme 2 (ACE2) interaction.
Methods: We developed methods and reagents to detect SARS-CoV-2 antibodies by enzyme-linked immunosorbent assay (ELISA). The main assays focus on the parallel detection of immunoglobulin (Ig)Gs against the spike trimer, its receptor binding domain (RBD) and nucleocapsid (N). We automated a surrogate neutralisation (sn)ELISA that measures inhibition of ACE2-spike or -RBD interactions by antibodies. The assays were calibrated to a World Health Organization reference standard.
Results: Our single-point IgG-based ELISAs accurately distinguished non-infected and infected individuals. For seroprevalence assessment (in a non-vaccinated cohort), classifying a sample as positive if antibodies were detected for ≥ 2 of the 3 antigens provided the highest specificity. In vaccinated cohorts, increases in anti-spike and -RBD (but not -N) antibodies are observed. We present detailed protocols for serum/plasma or dried blood spots analysis performed manually and on automated platforms. The snELISA can be performed automatically at single points, increasing its scalability.
Conclusions: Measuring antibodies to three viral antigens and identify neutralising antibodies capable of disrupting spike-ACE2 interactions in high-throughput enables large-scale analyses of humoral immune responses to SARS-CoV-2 infection and vaccination. The reagents are available to enable scaling up of standardised serological assays, permitting inter-laboratory data comparison and aggregation.
Keywords: SARS‐CoV‐2; antibody detection; antibody neutralisation; assay development and standardisation; high‐throughput screening.