University College London BMSC - Immunocovidome assay

SARS-CoV2 Immunocomplex by LC-MS/MS on vaccinated cohort
Data License: CC BY 4.0 | ProteomeXchange: PXD028450 | doi: https://doi.org/10.6069/36r1-3z82
  • Organism: Homo sapiens
  • Instrument: Xevo TQ-S
  • SpikeIn: No
  • Keywords: SARS-CoV2, Antibody, complement, spike
  • Lab head: Wendy Heywood Submitter: Wendy Heywood
Abstract
We describe the development of a highly specific, mulitplexed targeted proteomic LC-MS/MS assay for accurate quantitation of immunoprotective-proteins (including several classes of antibodies and complement) to SARS CoV2 S1 spike protein. We demonstrate how antibodies/complement levels can be monitored in response to vaccination with or without previous infection and show their altered level of protection against variants of concern.
Experiment Description
The experiment was performed on serum samples taken from volunteers receiving the Pfizer-BioNTech vaccine at first and second dose. Half of the participants had previous exposure from a SARS-CoV2 infection. Samples were incubated with a S1 spike protein bait derived from the Wuhan hu-1 original virus that vaccines are engineered against. S1 protein corresponding to the alpha, beta and delta variants of concern were also used. The bound immuncomplex was analysed by a mutlipex targeted LC-MS/MS assay for several antibody classes and associated complement factors.
Sample Description
Human sera were obtained from the COVIDsortium Healthcare Workers bioresource which is approved by the ethical committee of UK National Research Ethics Service (20/SC/0149) and registered on ClinicalTrials.gov (NCT04318314). The study conformed to the principles of the Helsinki Declaration, and all subjects gave written informed consent. COVIDsortium Healthcare Worker Participants: WT SARS-CoV-2 infection of study participants was determined using baseline and weekly nasal RNA stabilizing swabs and Roche cobas® SARS-CoV-2 reverse transcriptase polymerase chain reaction (RT-PCR) test as well as baseline and weekly antibody testing using the spike protein S1 IgG EUROIMMUN enzyme-linked immunosorbent assay (ELISA) and nucleocapsid ROCHE Elecsys electrochemiluminescence immunoassay (ECLIA). Antibody ratios >1.1 were considered test positive for the EUROIMMUN SARS-CoV-2 ELISA and >1 was considered test positive for the ROCHE Elecsys anti-SARS-CoV-2 ECLIA following Public Health England evaluation 1 The cross-sectional, case-controlled vaccine sub-study reported here (n = 51) collected samples at a mean/median timepoint of 22d (±2d SD) after administration of the first dose of the mRNA vaccine, BNT162b2 . This vaccine sub-study recruited HCW previously enrolled in the 16-18 week sub-study (11). This included 25 HCW (mean age 44yr, 60% male) with previous laboratory defined evidence of WT SARS-CoV-2 infection and twenty-six HCW (mean age 41y, 54% male) with no laboratory evidence of SARS-CoV-2 infection throughout the initial 16-week longitudinal follow up. Raw Data have participant sample ID and V1 to indicate first vaccine sample or V2 to indicate second vaccine. Other samples are pre vaccine. Processed exported and normalised data are provided in accompanying excel file.
Created on 9/13/21, 2:56 PM
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