tetano
Editor, Senior Moderator
Int Arch Allergy Immunol
. 2021 Feb 23;1-8.
doi: 10.1159/000514203. Online ahead of print.
Quantitative Measurement of IgG to Severe Acute Respiratory Syndrome Coronavirus-2 Proteins Using ImmunoCAP
Behnam Keshavarz[SUP] 1 [/SUP], Joesph R Wiencek[SUP] 2 [/SUP], Lisa J Workman[SUP] 1 [/SUP], Matthew D Straesser[SUP] 1 [/SUP], Lyndsey M Muehling[SUP] 1 [/SUP], Glenda Canderan[SUP] 1 [/SUP], Fabrizio Drago[SUP] 3 [/SUP], Catherine A Bonham[SUP] 4 [/SUP], Jeffrey M Sturek[SUP] 4 [/SUP], Chintan Ramani[SUP] 4 [/SUP], Coleen A McNamara[SUP] 3 [/SUP], Judith A Woodfolk[SUP] 1 [/SUP], Alexandra Kadl[SUP] 4 5 [/SUP], Thomas A E Platts-Mills[SUP] 1 [/SUP], Jeffrey M Wilson[SUP] 6 [/SUP]
Affiliations
Abstract
Background: Detailed understanding of the immune response to severe acute respiratory syndrome coronavirus (SARS-CoV)-2, the cause of coronavirus disease 2019 (CO-VID-19) has been hampered by a lack of quantitative antibody assays.
Objective: The objective was to develop a quantitative assay for IgG to SARS-CoV-2 proteins that could be implemented in clinical and research laboratories.
Methods: The biotin-streptavidin technique was used to conjugate SARS-CoV-2 spike receptor-binding domain (RBD) or nucleocapsid protein to the solid phase of the ImmunoCAP. Plasma and serum samples from patients hospitalized with COVID-19 (n = 60) and samples from donors banked before the emergence of COVID-19 (n = 109) were used in the assay. SARS-CoV-2 IgG levels were followed longitudinally in a subset of samples and were related to total IgG and IgG to reference antigens using an ImmunoCAP 250 platform.
Results: At a cutoff of 2.5 μg/mL, the assay demonstrated sensitivity and specificity exceeding 95% for IgG to both SARS-CoV-2 proteins. Among 36 patients evaluated in a post-hospital follow-up clinic, median levels of IgG to spike-RBD and nucleocapsid were 34.7 μg/mL (IQR 18-52) and 24.5 μg/mL (IQR 9-59), respectively. Among 17 patients with longitudinal samples, there was a wide variation in the magnitude of IgG responses, but generally the response to spike-RBD and to nucleocapsid occurred in parallel, with peak levels approaching 100 μg/mL, or 1% of total IgG.
Conclusions: We have described a quantitative assay to measure IgG to SARS-CoV-2 that could be used in clinical and research laboratories and implemented at scale. The assay can easily be adapted to measure IgG to mutated COVID-19 proteins, has good performance characteristics, and has a readout in standardized units.
Keywords: Antibody assay; COVID-19; IgG; Nucleocapsid; Spike receptor-binding domain.
. 2021 Feb 23;1-8.
doi: 10.1159/000514203. Online ahead of print.
Quantitative Measurement of IgG to Severe Acute Respiratory Syndrome Coronavirus-2 Proteins Using ImmunoCAP
Behnam Keshavarz[SUP] 1 [/SUP], Joesph R Wiencek[SUP] 2 [/SUP], Lisa J Workman[SUP] 1 [/SUP], Matthew D Straesser[SUP] 1 [/SUP], Lyndsey M Muehling[SUP] 1 [/SUP], Glenda Canderan[SUP] 1 [/SUP], Fabrizio Drago[SUP] 3 [/SUP], Catherine A Bonham[SUP] 4 [/SUP], Jeffrey M Sturek[SUP] 4 [/SUP], Chintan Ramani[SUP] 4 [/SUP], Coleen A McNamara[SUP] 3 [/SUP], Judith A Woodfolk[SUP] 1 [/SUP], Alexandra Kadl[SUP] 4 5 [/SUP], Thomas A E Platts-Mills[SUP] 1 [/SUP], Jeffrey M Wilson[SUP] 6 [/SUP]
Affiliations
- PMID: 33621972
- DOI: 10.1159/000514203
Abstract
Background: Detailed understanding of the immune response to severe acute respiratory syndrome coronavirus (SARS-CoV)-2, the cause of coronavirus disease 2019 (CO-VID-19) has been hampered by a lack of quantitative antibody assays.
Objective: The objective was to develop a quantitative assay for IgG to SARS-CoV-2 proteins that could be implemented in clinical and research laboratories.
Methods: The biotin-streptavidin technique was used to conjugate SARS-CoV-2 spike receptor-binding domain (RBD) or nucleocapsid protein to the solid phase of the ImmunoCAP. Plasma and serum samples from patients hospitalized with COVID-19 (n = 60) and samples from donors banked before the emergence of COVID-19 (n = 109) were used in the assay. SARS-CoV-2 IgG levels were followed longitudinally in a subset of samples and were related to total IgG and IgG to reference antigens using an ImmunoCAP 250 platform.
Results: At a cutoff of 2.5 μg/mL, the assay demonstrated sensitivity and specificity exceeding 95% for IgG to both SARS-CoV-2 proteins. Among 36 patients evaluated in a post-hospital follow-up clinic, median levels of IgG to spike-RBD and nucleocapsid were 34.7 μg/mL (IQR 18-52) and 24.5 μg/mL (IQR 9-59), respectively. Among 17 patients with longitudinal samples, there was a wide variation in the magnitude of IgG responses, but generally the response to spike-RBD and to nucleocapsid occurred in parallel, with peak levels approaching 100 μg/mL, or 1% of total IgG.
Conclusions: We have described a quantitative assay to measure IgG to SARS-CoV-2 that could be used in clinical and research laboratories and implemented at scale. The assay can easily be adapted to measure IgG to mutated COVID-19 proteins, has good performance characteristics, and has a readout in standardized units.
Keywords: Antibody assay; COVID-19; IgG; Nucleocapsid; Spike receptor-binding domain.