Multi plasmid system for the production of influenza virus
AssigneeMedImmune, LLC
InventorsGeorge Kemble, Gregory Duke
Life of this patent
Estimating the term…
Abstract
Vectors and methods for the production of influenza viruses suitable as recombinant influenza vaccines in cell culture are provided. Bi-directional expression vectors for use in a multi-plasmid influenza virus expression system are provided. Additionally, the invention provides methods of producing influenza viruses with enhanced ability to replicate in embryonated chicken eggs and/or cells (e.g., Vero and/or MDCK) and further provides influenza viruses with enhanced replication characteristics. In addition, the present invention includes an improved method of rescue, wherein animal cells (e.g., SF Vero cells) are electroporated with plasmids and vectors of the invention.
Claims17
A method for rescue of a reassortant influenza virus, comprising:
- (i) electroporating Vero cells with polynucleotide vectors that direct the expression in the Vero cells of genomic or antigenomic vRNA segments, a nucleoprotein and an RNA-dependent polymerase, whereby ribonucleoprotein complexes can be formed and viral particles can be assembled in the absence of helper virus;
- (ii) co-cultivating the electroporated Vero cells with another cell type, which cell type was not electroporated in step (i), under conditions permissive for viral replication;
- and
- (iii) recovering influenza virus, wherein the efficiency of the rescue is at least 90%.
Dependencies are inferred from the claim text.