Surface-displayed porcine reproductive and respiratory syndrome virus from cell culture onto gram-positive enhancer matrix particles

Abstract

Vaccine immunization is now one of the most effective ways to control porcine reproductive and respiratory syndrome virus (PRRSV) infection. Impurity is one of the main factors affecting vaccine safety and efficacy. Here we present a novel innovative PRRSV purification approach based on surface display technology. First, a bifunctional protein PA-GRFT (protein anchor-griffithsin), the crucial factor in the purification process, was successfully produced in Escherichia coli yielding 80 mg/L of broth culture. Then PRRSV purification was performed by incubation of PA-GRFT with PRRSV and gram-positive enhancer matrix (GEM) particles, followed by centrifugation to collect virions loaded onto GEM particles. Our results showed that most of the bulk impurities had been removed, and PA-GRFT could capture PRRSV onto GEM particles. Our lactic acid bacteria-based purification method, which is promising as ease of operation, low cost and easy to scale-up, may represent a candidate method for the large-scale purification of this virus for vaccine production.

Bacterium-Like Particles Displaying the Rift Valley Fever Virus Gn Head Protein Induces Efficacious Immune Responses in Immunized Mice

“…In recent years, a novel antigen delivery system consisting of non-living, non-genetically modified cell wall particles derived from the food-grade bacterium Lactococcus lactis MG1363 is described for a number of pathogens, including shigellosis ( Heine et al, 2015 ), human papillomavirus (HPV) ( Ribelles et al, 2013 ), influenza virus (H1N1) ( Saluja et al, 2010 ), porcine circovirus type 2 (PCV2) ( Qiao et al, 2019 ), hepatitis E virus (HEV) ( Gao et al, 2015 ), foot-and-mouth disease virus (FMDV) ( Cheng et al, 2019 ), severe acute respiratory syndrome coronavirus (SARS) ( Lee et al, 2006 ), and porcine reproductive and respiratory syndrome virus (PRRSV) ( Li et al, 2018 ). This novel antigen delivery system allows for the display of multiple antigens on the particle surface in the form of recombinant fusion proteins containing a heterogeneous antigen and a peptidoglycan protein anchor (PA).…”

Immune efficacy of a porcine circovirus type 2 vaccine purified using Gram‐positive enhancer matrix surface display technology

“…However, few studies have demonstrated that GEM particles can display complete virions cultured in cell lines. In order to solve this problem, another PL protein (PA‐GRFT) was used to purify PRRSV (Li et al ). However, GRFT only combined with PRRSV and was not available for PCV2.…”

Bacterium-like particles derived from probiotics: progress, challenges and prospects

“…Further experimental results demonstrated that the PA-GRFT partnership could guide PRRSV to the BLPs surface, yielding favorable results in terms of efficiency (64). Additional research using this process to purify additional enveloped animal viruses may result in a brand-new, broadly applicable technique for viral purification (65). Another typical study connected foot-and-mouth disease virus (FMDV)-specific nanobody to BLPs through PA, named GEM-PA-nanotrap, was used to FMDV purification from cellular lysates.…”