In:
PLOS ONE, Public Library of Science (PLoS), Vol. 17, No. 5 ( 2022-5-24), p. e0268767-
Abstract:
Since the outbreak of the COVID-19 pandemic, widespread infections have allowed SARS-CoV-2 to evolve in human, leading to the emergence of multiple circulating variants. Some of these variants show increased resistance to vaccine-elicited immunity, convalescent plasma, or monoclonal antibodies. In particular, mutations in the SARS-CoV-2 spike have drawn attention. To facilitate the isolation of neutralizing antibodies and the monitoring of vaccine effectiveness against these variants, we designed and produced biotin-labeled molecular probes of variant SARS-CoV-2 spikes and their subdomains, using a structure-based construct design that incorporated an N-terminal purification tag, a specific amino acid sequence for protease cleavage, the variant spike-based region of interest, and a C-terminal sequence targeted by biotin ligase. These probes could be produced by a single step using in-process biotinylation and purification. We characterized the physical properties and antigenicity of these probes, comprising the N-terminal domain (NTD), the receptor-binding domain (RBD), the RBD and subdomain 1 (RBD-SD1), and the prefusion-stabilized spike ectodomain (S2P) with sequences from SARS-CoV-2 variants of concern or of interest, including variants Alpha, Beta, Gamma, Epsilon, Iota, Kappa, Delta, Lambda, Mu, and Omicron. We functionally validated probes by using yeast expressing a panel of nine SARS-CoV-2 spike-binding antibodies and confirmed sorting capabilities of variant probes using yeast displaying libraries of plasma antibodies from COVID-19 convalescent donors. We deposited these constructs to Addgene to enable their dissemination. Overall, this study describes a matrix of SARS-CoV-2 variant molecular probes that allow for assessment of immune responses, identification of serum antibody specificity, and isolation and characterization of neutralizing antibodies.
Type of Medium:
Online Resource
ISSN:
1932-6203
DOI:
10.1371/journal.pone.0268767
DOI:
10.1371/journal.pone.0268767.g001
DOI:
10.1371/journal.pone.0268767.g002
DOI:
10.1371/journal.pone.0268767.g003
DOI:
10.1371/journal.pone.0268767.g004
DOI:
10.1371/journal.pone.0268767.g005
DOI:
10.1371/journal.pone.0268767.g006
DOI:
10.1371/journal.pone.0268767.g007
DOI:
10.1371/journal.pone.0268767.g008
DOI:
10.1371/journal.pone.0268767.s001
DOI:
10.1371/journal.pone.0268767.s002
DOI:
10.1371/journal.pone.0268767.s003
DOI:
10.1371/journal.pone.0268767.s004
DOI:
10.1371/journal.pone.0268767.s005
DOI:
10.1371/journal.pone.0268767.s006
DOI:
10.1371/journal.pone.0268767.s007
DOI:
10.1371/journal.pone.0268767.s008
DOI:
10.1371/journal.pone.0268767.s009
DOI:
10.1371/journal.pone.0268767.s010
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2022
detail.hit.zdb_id:
2267670-3
Permalink