A HiBiT-tagged pseudovirus-like particle platform for safe, rapid quantification of virus neutralization and antibody-dependent enhancement

Standard neutralization assays are often slow, labor-intensive, and restricted to high-containment facilities, thus complicating and delaying the development of vaccines and antibody-based treatments. Here, we present a novel neutralization assay system using HiBiT-tagged pseudovirus-like particles (HiBiT-PsVLPs). These particles incorporate entry proteins from diverse pathogenic viruses but are non-replicating and lack viral nucleic acids, thus mitigating the biosafety risks of conventional assays. The particles encapsulate the HiBiT peptide, enabling rapid, luminescent quantitation of entry and neutralization. We demonstrate that this platform accurately measures neutralizing activity of monoclonal antibodies across development stages and sensitively detects antibody-dependent enhancement, a critical safety consideration. Altogether, HiBiT-PsVLPs offer a safe, rapid, and scalable platform to accelerate the development of vaccines and antibody therapeutics targeting a broad range of viruses.