Selection free growth of host cells containing multiple integrating vectors
AssigneeGala Design, Inc.
InventorsRobert D. Bremel, Gregory T. Bleck, Dona York, Kurt Eakle
Life of this patent
Estimating the term…
Abstract
The present invention relates to the production of proteins in host cells, and more particularly to host cells containing multiple integrated copies of an integrating vector comprising an exogenous gene. The present invention further relates to the use of integrating vectors lacking a selectable marker and growth of host cells containing such vectors in the absence of selection. The present invention further provides methods of expressing increased levels of protein in host cells using such vectors.
Claims50
A host cell comprising a genome, said genome comprising at least one integrated integrating vector, wherein said integrating vector comprises at least one exogenous gene operably linked to a promoter, and wherein said integrating vectors lacks a gene encoding a selectable marker.
A method for transfecting host cells comprising:
- a) providing:
- i) a plurality of host cells comprising a genome, and
- ii) a plurality of integrating vectors, wherein said integrating vectors comprise at least one exogenous gene, and wherein said integrating vectors lack a gene encoding a selectable marker;
- b) contacting said host cell with said plurality of integrating vectors to generate transfected host cells comprising at least one integrated copy of said integrating vector;
- and
- c) clonally selecting said transfected host cells.
A method of producing a protein of interest comprising:
- a) providing a host cell comprising a genome, said genome comprising at least one integrated copy of at least one integrating vector comprising an exogenous gene operably linked to a promoter, wherein said integrating vector lacks a gene encoding a selectable marker, and wherein said exogenous gene encodes a protein of interest, and
- b) culturing said host cells under conditions such that said protein of interest is produced.
A retroviral vector comprising a gene construct comprising an exogenous promoter operably linked to an exogenous gene, said vector lacking a gene encoding a selectable marker.
Dependencies are inferred from the claim text.