Template-Assisted De Novo Sequencing of SARS-CoV-2 and Influenza Monoclonal Antibodies by Mass Spectrometry

• DOI: 10.1021/acs.jproteome.1c00913
• Publication Date: 2022-06-02T21:42:35Z
• Authors: Michelle V. Gadush; Giuseppe A. Sautto; Hamssika Chandras...; Alena Bensussan; Ted M. Ross; Gregory C. Ippolito; Maria D. Person
• Keywords: 0303 health sciences; 03 medical and health sciences; SARS-CoV-2; Influenza, Human; Antibodies, Monoclonal; COVID-19; Humans; Antibodies, Viral; Mass Spectrometry

In this study, we used multiple enzyme digestions, coupled with higher-energy collisional dissociation (HCD) and electron-transfer/higher-energy collision (EThcD) fragmentation to develop a mass-spectrometric (MS) method for determining the complete protein sequence of monoclonal antibodies (mAbs). The was refined on an mAb known sequence, SARS-CoV-1 antireceptor binding domain (RBD) spike antibody. data were searched using Supernovo generate template-assisted de novo two SARS-CoV-2 mAbs sequences resulting in correct variable regions distinction Ile Leu residues. We then set 25 antihemagglutinin (HA) influenza unknown determined high confidence >99% complementarity (CDRs). heavy-chain light-chain genes cloned transfected into cells recombinant expression followed by affinity purification. displayed curves matching original specificity HA antigen. Our findings indicate that methodology results almost antibody coverage CDR diverse sequences.