| Autor: |
Fisher M; Department of Infectious Diseases, Israel Institute for Biological Research, 7410001 Ness-Ziona, Israel., Manor A; Department of Environmental Physics, Israel Institute for Biological Research, 7410001 Ness Ziona, Israel., Abramovitch H; Department of Quality Assurance, Israel Institute for Biological Research, 7410001 Ness Ziona, Israel., Fatelevich E; Department of Infectious Diseases, Israel Institute for Biological Research, 7410001 Ness-Ziona, Israel., Afrimov Y; Department of Infectious Diseases, Israel Institute for Biological Research, 7410001 Ness-Ziona, Israel., Bilinsky G; Department of Biochemistry and Molecular Genetics, Israel Institute for Biological Research, 7410001 Ness Ziona, Israel., Lupu E; Department of Biotechnology, Israel Institute for Biological Research, 7410001 Ness Ziona, Israel., Ben-Shmuel A; Department of Infectious Diseases, Israel Institute for Biological Research, 7410001 Ness-Ziona, Israel., Glinert I; Department of Infectious Diseases, Israel Institute for Biological Research, 7410001 Ness-Ziona, Israel., Madar-Balakirski N; Department of Pharmacology, Israel Institute for Biological Research, 7410001 Ness Ziona, Israel., Marcus H; Department of Biotechnology, Israel Institute for Biological Research, 7410001 Ness Ziona, Israel., Mechaly A; Department of Infectious Diseases, Israel Institute for Biological Research, 7410001 Ness-Ziona, Israel. |
| Abstrakt: |
A multi-component microarray, applying a novel analysis algorithm, was developed for quantitative evaluation of the SARS-CoV-2 vaccines' immunogenicity. The array enables simultaneous quantitation of IgG, IgM, and IgA, specific to the SARS-CoV-2 spike, receptor binding domain, and nucleocapsid proteins. The developed methodology is based on calculating an apparent immunoglobulin signal from the linear range of the fluorescent read-outs generated by scanning the microarray slides at different exposure times. A dedicated algorithm, employing a rigorous set of embedded conditions, then generates a normalized signal for each of the unique assays. Qualification of the multi-component array performance (evaluating linearity, extended dynamic-range, specificity, precision, and accuracy) was carried out with an in-house COVID-19, qRT-PCR positive serum, as well as pre-pandemic commercial negative sera. Results were compared to the WHO international standard for anti-SARS-CoV-2 immunoglobulins. Specific IgG, IgM, and IgA signals obtained by this array enabled successful discrimination between SARS-CoV-2 q-RT-PCR positive (seroconverted SARS-CoV-2 patients) and negative (naïve) samples. This array is currently used for evaluation of the humoral response to BriLife, the VSV-based Israeli vaccine during phase I/II clinical trials. |
