Detection of Neutralization-sensitive Epitopes in Antigens Displayed on Virus-Like Particle (VLP)-Based Vaccines Using a Capture Assay

The virus-like particle (VLP) capture assay is an immunoprecipitation method, commonly known as a 'pull-down assay' used to purify and isolate antigen-displaying VLPs. Surface antigen-specific antibodies are coupled to, thus immobilized on solid insoluble matrix such beads. Due their high affinity the target antigen, these can VLPs decorated with cognate antigen anchored in membrane envelope of This protocol describes binding protein A- or G-conjugated magnetic In our study, human immunodeficiency virus (HIV)-derived formed by group-specific (Gag) viral core precursor p55 Gag displaying glycoproteins (Env) HIV examined. captured utilizing broadly neutralizing (bNAbs) directed against neutralization-sensitive epitopes Env. VLP outlined here represents sensitive easy-to-perform method demonstrate that (i) respective (ii) surface retained its structural integrity demonstrated epitope-specific bNAbs (iii) revealed detection proteins subsequent Western blot-analysis. Consequently, utilization for facilitates prediction whether vaccines will be able elicit B cell response vaccinated humans. We anticipate this furnish other researchers valuable straightforward experimental approach examine potential VLP-based vaccines.