» Articles » PMID: 39192985

A Single Domain Antibody-based Luminex Assay for the Detection of SARS-CoV-2 in Clinical Samples

Overview
Journal Front Immunol
Date 2024 Aug 28
PMID 39192985
Authors
Affiliations
Soon will be listed here.
Abstract

Within the past decade, single domain antibodies (sdAbs) have been recognized as unique affinity binding reagents that can be tailored for performance in a variety of immunoassay formats. Luminex MagPlex color-coded magnetic microspheres provide a high-throughput platform that enables multiplexed immunoassays. We developed a MagPlex bead-based assay for the detection of SARS-CoV-2, using sdAbs against SARS-CoV-2 nucleocapsid (N) protein in which we engineered the sdAb capture reagents to orient them on the beads. The oriented sdAbs provided an increase in sensitivity over randomly oriented sdAbs for samples of N diluted in buffer, which also translated into better detection of SARS-CoV-2 in clinical samples. We assessed the specificity of the assay by examining seasonal coronavirus clinical samples. In summary, we provide a proof-of-concept that a bead-based assay using sdAbs to detect SARS-CoV-2 is feasible and future research combining it with other sdAb-coated beads that can detect other viruses may provide a useful diagnostic tool.

References
1.
Lu R, Zhao X, Li J, Niu P, Yang B, Wu H . Genomic characterisation and epidemiology of 2019 novel coronavirus: implications for virus origins and receptor binding. Lancet. 2020; 395(10224):565-574. PMC: 7159086. DOI: 10.1016/S0140-6736(20)30251-8. View

2.
Wu C, Qavi A, Hachim A, Kavian N, Cole A, Moyle A . Characterization of SARS-CoV-2 nucleocapsid protein reveals multiple functional consequences of the C-terminal domain. iScience. 2021; 24(6):102681. PMC: 8168301. DOI: 10.1016/j.isci.2021.102681. View

3.
Pollock N, Savage T, Wardell H, Lee R, Mathew A, Stengelin M . Correlation of SARS-CoV-2 Nucleocapsid Antigen and RNA Concentrations in Nasopharyngeal Samples from Children and Adults Using an Ultrasensitive and Quantitative Antigen Assay. J Clin Microbiol. 2021; 59(4). PMC: 8092747. DOI: 10.1128/JCM.03077-20. View

4.
Zavrtanik U, Lukan J, Loris R, Lah J, Hadzi S . Structural Basis of Epitope Recognition by Heavy-Chain Camelid Antibodies. J Mol Biol. 2018; 430(21):4369-4386. DOI: 10.1016/j.jmb.2018.09.002. View

5.
Xu J, Kerr L, Jiang Y, Suo W, Zhang L, Lao T . Rapid Antigen Diagnostics as Frontline Testing in the COVID-19 Pandemic. Small Sci. 2022; 2(8):2200009. PMC: 9349911. DOI: 10.1002/smsc.202200009. View