Investigation of Kluyveromyces marxianus as a novel host for large‐scale production of porcine parvovirus virus‐like particles

Abstract Background Porcine Parvovirus (PPV) is a Parvovirinae virus that can cause embryonic and fetal loss death mummification in affected pigs. Unlike conventional vaccines, virus-like particles (VLPs) inherit the natural structure of their authentic virions highly immunostimulatory induce strong humoral immune T cell responses with no risk pathogenicity. The production PPV VLPs still challenge based on traditional expression platforms due to low yields high culture costs. Kluyveromyces marxianus safe fast-growing eukaryote get biomass low-cost cultures. In this study, we investigated downstream processes K. , potential for effective stand-alone vaccines. Results After optimization according codon bias VP2 protein from Kresse strain was expressed. 5 L fermentator, yield reached 2.5 g/L, quantified by HPLC, using defined mineral medium after 48 h fermentation. Two strategies were established purify intracellular VLPs: (i) Using cation exchange chromatography coupled Sephacryl® S-500 HR supernatants pH adjusted lysates. (ii) anion followed cross-flow diafiltration recover precipitated purity about 95%, total recovery more than 60%. Vaccination mice purified induced titers specific IgG antibodies sera, showed hemagglutination inhibitions both swine guinea pig erythrocytes. Spleen lymphocyte proliferation cytokines detection suggested produced provoked cellular immunity mice. Conclusions This highest recombinant achieved date. superiorities, Generally Recognized As Safe (GRAS), production, short lead time, cost, make greatly competitive platform bioproduction vaccine.