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  • Next-Generation Serology by Mass Spectrometry: Readout of the SARS-CoV-2 Antibody Repertoire.

Next-Generation Serology by Mass Spectrometry: Readout of the SARS-CoV-2 Antibody Repertoire.

Journal of proteome research (2021-12-09)
Rafael D Melani, Benjamin J Des Soye, Jared O Kafader, Eleonora Forte, Michael Hollas, Voislav Blagojevic, Fernanda Negrão, John P McGee, Bryon Drown, Cameron Lloyd-Jones, Henrique S Seckler, Jeannie M Camarillo, Philip D Compton, Richard D LeDuc, Bryan Early, Ryan T Fellers, Byoung-Kyu Cho, Basil Baby Mattamana, Young Ah Goo, Paul M Thomas, Michelle K Ash, Pavan P Bhimalli, Lena Al-Harthi, Beverly E Sha, Jeffrey R Schneider, Neil L Kelleher
ABSTRACT

Methods of antibody detection are used to assess exposure or immunity to a pathogen. Here, we present Ig-MS, a novel serological readout that captures the immunoglobulin (Ig) repertoire at molecular resolution, including entire variable regions in Ig light and heavy chains. Ig-MS uses recent advances in protein mass spectrometry (MS) for multiparametric readout of antibodies, with new metrics like Ion Titer (IT) and Degree of Clonality (DoC) capturing the heterogeneity and relative abundance of individual clones without sequencing of B cells. We applied Ig-MS to plasma from subjects with severe and mild COVID-19 and immunized subjects after two vaccine doses, using the receptor-binding domain (RBD) of the spike protein of SARS-CoV-2 as the bait for antibody capture. Importantly, we report a new data type for human serology, that could use other antigens of interest to gauge immune responses to vaccination, pathogens, or autoimmune disorders.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Human IgG4 Kappa, Recombinant Protein, purified immunoglobulin, buffered aqueous solution
Sigma-Aldrich
IgM from human serum, reagent grade, ~95% (HPLC), buffered aqueous solution
Sigma-Aldrich
Goat Anti-Rabbit IgG Antibody, (H+L) HRP conjugate, 1 mg/mL, Chemicon®
Sigma-Aldrich
IgA from human serum, reagent grade, ≥95% (HPLC), buffered aqueous solution