UCLA researchers in the Department of Microbiology, Immunology, and Molecular Genetics have developed a matrix based virus-like particle entry and budding assay that can be used to study highly pathogenic viruses at lower containment conditions. The optimized biologically relevant assay is sensitive, specific and allows for easy quantitative detection of viral particles
Many viral entry studies on highly pathogenic agents rely on cell-cell fusion and envelope pseudotyped reporter assays. These assays allow for detailed analyses of virus entry characteristics without high-level biosafety containment. Unfortunately, these surrogate assays may not fully emulate the biological properties of native envelope structures that are unique to the virus being studied.
Researchers at UCLA have developed a matrix based virus-like particle entry and budding assay that overcomes the limitations of surrogate assays and offers significant advantages, namely: extreme sensitivity, adaptability to other viruses with matrix determinants, and immediate entry detection without the requirement for reporter gene transcription/translations. The optimized entry and budding assay is based on a β-lactamase-Nipah matrix fusion protein which is able to bud and form virus-like particles that morphologically resemble paramyxoviruses. To increase detectability, the researchers have also created mammalian codon-optimized and catalytically improved versions of the β-lactamase matrix fusion genes. The matrix based virus-like particle assay can be used to study highly pathogenic (BSL-4) viruses under BSL-2 conditions. In addition, the codon-optimized β-lactamase matrix fusion genes can be used for efficient production of virus-like particles for vaccines.
The virus-like particle entry and budding assays have been established for Nipah and Hendra viruses. The assay has been optimized with a β-lactamase mutation, as well as mammalian codon-optimization of both genes in the fusion construct.
research tool, virus, vaccine production, virus-like particle, VLP, henipavirus, paramyxovirus, Nipah, Hendra, virus entry and budding assay, cell-cell fusion, envelope pseudotyped reporter assays, beta-lactamase