High-yield Transgenic Mammalian Expression System for Generating Virus-like Particles
AssigneeAcademia Sinica
InventorsPei-Wen Hsiao, Chia- Ying Wu, Yi-Chun Yeh
Life of this patent
Estimating the term…
Abstract
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.
Claims26
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.
A method of preparing an influenza virus-like particle (VLP), the method comprising:
- obtaining a co-expression Vero cell stably transfected with a sequence encoding an influenza M1 and a sequence encoding an influenza M2, and further transfected with a sequence encoding an influenza hemagglutinin (HA) and a sequence encoding an influenza neuraminidase (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;
- and
- 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
An influenza virus-like particle (VLP), comprising:
- an influenza M1, an influenza M2, an influenza hemagglutinin (HA) and an influenza neuraminidase (NA), wherein the influenza proteins are recombinantly expressed from a Vero cell;
- and
- at least one cellular protein of the Vero cell.
A founder Vero cell being a Vero cell stably transfected with a sequence encoding an influenza M1 and a sequence encoding an influenza M2, wherein the expressions of the influenza M1 and M2 proteins in the founder Vero cell are controlled by an inducible expression system.
A method of obtaining a founder Vero cell, the method comprising:
- introducing into a Vero cell a sequence encoding an influenza M1 and a sequence encoding an influenza M2;
- and
- obtaining the founder Vero cell stably transfected with the sequence encoding the influenza M1 and the sequence encoding the influenza M2,
- wherein the expressions of the influenza M1 and M2 proteins in the founder Vero cell are controlled by an inducible expression system.
Dependencies are inferred from the claim text.