HIGH-YIELD TRANSGENIC MAMMALIAN EXPRESSION SYSTEM FOR GENERATING VIRUS-LIKE PARTICLES

摘要

Virus-like particles (VLPs) of mammalian-hosted viruses, such as SARS-CoV and influenza viruses, have been recombinantly produced from Vero cells. The VLPs closely emulate the exterior of authentic virus particles and are highly immunogenic. They can elicit not only humoral but also cellular immune responses in a mammal. Compositions and methods related to the VLPs are also described.

主权项

1. A method of preparing an influenza virus-like particle (VLP), the method comprising:
obtaining a founder Vero cell stably transfected with a sequence encoding an influenza M1 and a sequence encoding an influenza M2; constructing at least one recombinant DNA molecule comprising a sequence encoding an influenza hemagglutinin (HA) and a sequence encoding an influenza neuraminidase (NA); introducing the at least one recombinant DNA molecule into the founder Vero cell to obtain a co-expression Vero cell stably transfected with the sequences encoding the influenza M1 and the influenza M2, and further transfected with the sequences encoding the influenza HA and the influenza NA, wherein the expressions of the influenza M1, M2, HA and NA proteins in the co-expression Vero cell are controlled by one or more inducible expression systems; culturing the co-expression Vero cell under conditions to allow expressions of the influenza M1, the influenza M2, the influenza HA and the influenza NA, and assembly of the VLP comprising the influenza M1, the influenza M2, the influenza HA and the influenza NA; and isolating the VLP from the culture supernatant of the co-expression Vero cell.