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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.

medRxiv : the preprint server for health sciences (2021-07-17)
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 multi-parametric 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 apply Ig-MS to plasma from subjects with severe & mild COVID-19, 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, with compatibility to any recombinant antigen to gauge our immune responses to vaccination, pathogens, or autoimmune disorders.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
IgA 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®