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

Abstract

Aims: Purification of porcine circovirus type 2 (PCV2) using Gram-positive enhancer matrix (GEM) surface display technology and immunogenicity evaluation of the purified antigen. Methods and Results: A recombinant bifunctional protein containing a protein anchor domain and a 'virus anchor' domain was designed as a protein linker (PL) between PCV2 and GEM particles. By incubating with PL and GEM particles sequentially, PCV2 could be purified and enriched through a simple centrifugation process with GEM surface display technology. Our data showed that one unit (2Á5 9 10 9 particles) of GEM particles with 80 lg PL could purify 100 ml of PCV2-containing culture supernatant (viral titre: 10 6Á5 TCID 50 per ml À1 ) with a recovery rate up to 99Á6%. The impurity removal efficiency of this method, calculated according to decreased total protein content during purification, was approximately 98%. Furthermore, in vivo experimentation showed that piglets immunized with purified PCV2 could elicit strong immune responses to prevent against PCV2 infection. Conclusion: Porcine circovirus type 2 could be efficiently purified and enriched with GEM display technology via a crucial PL, and the purified PCV2 could elicit effective immune responses against PCV2 infection. Significance and Impact of the Study: The GEM-based purification method established here is cost-efficient and high-throughput, and may represent a promising large-scale purification method for PCV2 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).…”

Cell Surface Display of Thermomyces lanuginosus Lipase in Pichia pastoris

“…Cell surface display integrates expression and immobilization of the enzyme together thus there is no requirement of complicated purification process except fusing the interest protein with anchor proteins through the convenient genetic operation (Kondo and Ueda, 2004). It has been used in many fields, such as live vaccine development, library screening and bioconversion (Georgiou et al, 1997;Tateno et al, 2007;Qiao et al, 2019), and have become a powerful technique in recent years. Up to now, few reports on the yeast surface display of TLL have been published, except Dai et al (2012).…”