A rapid bead-based assay for screening of SARS-CoV-2 neutralizing antibodies.
Antib Ther
; 5(2): 100-110, 2022 Apr.
Article
in English
| MEDLINE | ID: covidwho-1795393
ABSTRACT
Quantitative determination of neutralizing antibodies against Severe Acute Respiratory Syndrome Corona Virus-2 (SARS-CoV-2) is paramount in immunodiagnostics, vaccine efficacy testing, and immune response profiling among the vaccinated population. Cost-effective, rapid, easy-to-perform assays are essential to support the vaccine development process and immunosurveillance studies. We describe a bead-based screening assay for S1-neutralization using recombinant fluorescent proteins of hACE2 and SARS-CoV2-S1, immobilized on solid beads employing nanobodies/metal-affinity tags. Nanobody-mediated capture of SARS-CoV-2-Spike (S1) on agarose beads served as the trap for soluble recombinant ACE2-GFPSpark, inhibited by neutralizing antibody. The first approach demonstrates single-color fluorescent imaging of ACE2-GFPSpark binding to His-tagged S1-Receptor Binding Domain (RBD-His) immobilized beads. The second approach is dual-color imaging of soluble ACE2-GFPSpark to S1-Orange Fluorescent Protein (S1-OFPSpark) beads. Both methods showed a good correlation with the gold standard pseudovirion assay and can be adapted to any fluorescent platforms for screening.
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Topics:
Vaccines
Language:
English
Journal:
Antib Ther
Year:
2022
Document Type:
Article
Affiliation country:
Abt
Similar
MEDLINE
...
LILACS
LIS