Quantitative measurement of IgG to SARS-CoV-2 proteins using ImmunoCAP.
| Autor: | Keshavarz B, Wiencek JR, Workman LJ, Straesser MD, Muehling LM, Canderan G, Drago F, Bonham CA, Sturek JM, Ramani C, McNamara CA, Woodfolk JA, Kadl A, Platts-Mills TAE, Wilson JM |
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| Jazyk: | angličtina |
| Zdroj: | MedRxiv : the preprint server for health sciences [medRxiv] 2020 Nov 12. Date of Electronic Publication: 2020 Nov 12. |
| DOI: | 10.1101/2020.11.09.20228411 |
| Abstrakt: | Background: Detailed understanding of the immune response to SARS-CoV-2, the cause of coronavirus disease 2019 (COVID-19), has been hampered by a lack of quantitative antibody assays. Objective: To develop a quantitative assay for IgG to SARS-CoV-2 proteins that could readily 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 resin. Plasma and serum samples from patients with COVID-19 (n=51) and samples from donors banked prior to 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: Performance characteristics demonstrated 100% sensitivity and 99% specificity at a cut-off level of 2.5 µg/mL for 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 novel antigens, has good performance characteristics and a read-out in standardized units. |
| Databáze: | MEDLINE |
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