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) for a target influenza strain, the method comprising:
(1) obtaining a founder Vero cell, wherein the founder Veto cell is stably transfected with a sequence encoding an influenza M1 and a sequence encoding an influenza M2 comprising the amino acid sequence of SEQ ID NO: 7; (2) constructing at least one recombinant DNA molecule comprising a sequence encoding an influenza hemagglutinin (HA) and a sequence encoding an influenza neuraminidase (NA), wherein the influenza HA and NA are derived from the target influenza strain; (3) transfecting the founder Vero cell from step (1) with the at least one recombinant DNA molecule from step (2) to obtain a co-expression Vero cell, wherein the co-expression Vero cell expresses the influenza M1, M2, HA and NA proteins from one or more inducible expression systems; (4) culturing the co-expression Vero cell from step (3) under conditions for expressions of the influenza M1, M2, HA and NA proteins, assembly of the influenza VLP, and secretion of the influenza VLP into culture supernatant of the co-expression Vero cell; and (5) isolating the influenza VLP from the culture supernatant of the co-expression Vero cell, wherein the influenza VLP comprises the influenza M1, M2, HA and NA proteins and at least one cellular protein of the Vero cell, and the administration of the VLP to a subject induces an immunity against the target influenza strain in the subject.