Antibody Profiling and Prevalence in the US population during the SARS-CoV2 Pandemic

Abstract

Background: Antibody diagnostics play an important role in disease detection and can potentially aid in monitoring of the immune responses to see if an individual has developed immunity. Developing high throughput diagnostics which does not involve handling of infectious material becomes imperative in the case of pandemics such as the recent outbreak of SARS-CoV2. Methods:A protein microarray technology was used to detect the plurality of antibody response to four novel antigens namely S1 glycoprotein, Receptor binding domain (RBD), S2 glycoprotein and Nucleoprotein of the novel coronavirus named SARS-CoV2 using serum samples. A DBS card was additionally used to compare its performance with a venipuncture-based serum separator tube (SST) draw. Results:The three main subclasses of antibodies IgM, IgA and IgG were analyzed to see the variations in immune responses in the affected population and compared to their microbial RT-PCR based NP swab results. The clinical sensitivity and specificity were determined to be 98.1% and 98.6%. In the matrix comparison study, which would enable patients to test without risk of transmitting the virus, DBS matched with higher than 98% accuracy to a venipuncture-based SST collection. Conclusion:Multiplex testing enables higher sensitivity and specificity which is essential while establishing exposure on a population scale. This flexible platform along with a discrete collection methodology would be crucial and broadly useful to scale up testing in current and future pandemics.Minimum sample volume that can be collected using DBS cards can be processed in this multiplex pillar plate format enabling the capacity to provide the reliability of high throughput analyzers while having the ease of collection similar to rapid tests.

SARS-CoV2 envelop proteins reshape the serological responses of COVID-19 patients

“…Antibody production was detected against peptides derived from the structural proteins Spike (S1, S2 and RBD domains), N and M and from the accessory protein ORF3a. Interestingly, we are able to demonstrate for the first time the occurrence of an antibody response in COVID+ patients against the entire M protein using our cell-based serological assay, thereby confirming results observed in proteome microarrays in a more physiological context (20)(21)(22). Higher Ig binding was observed in hospitalized COVID+ patients with moderate and severe forms of the disease.…”

“…Recent studies have mapped the regions of SARS-CoV-2 proteins recognized by antibodies from COVID-19 patients using proteome microarrays (20)(21)(22). Antibody production was detected against peptides derived from the structural proteins Spike (S1, S2 and RBD domains), N and M and from the accessory protein ORF3a.…”

Identification of immunodominant linear epitopes from SARS-CoV-2 patient plasma

“…The rapid propagation of SARS-CoV-2 stimulated several studies predicting the antigenic parts of the viral proteins in silico [26][27][28][29][30][31][32], and analyzing SARS-CoV-1 epitopes that were conserved in this new coronavirus [33][34][35][36]. More recently, the first reports of experimental epitope mapping of the SARS-CoV-2 were deposited on repositories [37][38][39][40][41][42].…”

An ultra-high-density protein microarray for high throughput single-tier serological detection of Lyme disease

“…1d-g. The individual wafers for each antigen were then diced into 0.7 × 0.7 mm 2 microchips using a stealth dicing process, as previously described 21 . The diced wafers were picked and placed onto individual carrier tapes using a standard die sorting system.…”

“…Data analysis. The generated images are analyzed using the in-house reporter software as described previously 21 . Briefly, the chemiluminescent signals are converted into intensity plots which was compared to the thresholds established for each antigen to determine sero-positivity.…”