Earnest-Silveira L, Christiansen D, Herrmann S, Ralph S A, Das S, Gowans E J, Torresi J
Department of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, Victoria 3010, Australia.
Department of Microbiology and Immunology, The Peter Doherty Institute for Infection and Immunity, University of Melbourne, Parkville, Victoria 3010, Australia; Department of Surgery, Austin Hospital, University of Melbourne, Australia.
J Virol Methods. 2016 Oct;236:87-92. doi: 10.1016/j.jviromet.2016.06.012. Epub 2016 Jun 29.
A method for the large-scale production of a quadrivalent mammalian cell derived hepatitis C virus-like particles (HCV VLPs) is described. The HCV core E1 and E2 coding sequences of genotype 1a, 1b, 2a or 3a were co-expressed in Huh7 cell factories using a recombinant adenoviral expression system. The structural proteins self-assembled into VLPs that were purified from Huh7 cell lysates by iodixanol ultracentrifugation and Stirred cell ultrafiltration. Electron microscopy, revealed VLPs of the different genotypes that are morphologically similar. Our results show that it is possible to produce large quantities of individual HCV genotype VLPs with relative ease thus making this approach an alternative for the manufacture of a quadrivalent mammalian cell derived HCV VLP vaccine.
描述了一种大规模生产四价哺乳动物细胞衍生丙型肝炎病毒样颗粒（HCV VLPs）的方法。使用重组腺病毒表达系统在Huh7细胞工厂中共表达1a、1b、2a或3a基因型的HCV核心E1和E2编码序列。结构蛋白自组装成VLPs，通过碘克沙醇超速离心和搅拌细胞超滤从Huh7细胞裂解物中纯化。电子显微镜显示不同基因型的VLPs在形态上相似。我们的结果表明，可以相对轻松地大量生产单个HCV基因型VLPs，从而使这种方法成为生产四价哺乳动物细胞衍生HCV VLP疫苗的一种替代方法。
