Methods for making recombinant proteins using apoptosis inhibitors
AssigneeGenentech, Inc.
InventorsVishva Dixit, Robert W. Hamilton, Jana van de Goor
Life of this patent
Estimating the term…
Abstract
The invention provided improved methods of making and producing recombinant proteins in in vitro cultures of host cells using apoptosis inhibitors. The use of one or more apoptosis inhibitors in the methods can reduce apoptosis in the cell cultures and markedly improve yield of the desired recombinant proteins.
Claims37
A method of making recombinant proteins using one or more apoptosis inhibitors, comprising the steps of:
- (a) providing a vector comprising a gene encoding an apoptosis inhibitor,
- (b) providing a vector comprising a gene encoding a protein of interest,
- (c) providing a host cell,
- (d) transforming or transfecting the host cell with the vector of steps (a) and (b),
- (e) providing cell culture media,
- (f) culturing the transformed or transfected host cell in the cell culture media under conditions sufficient for expression of the protein of interest and the apoptosis inhibitor, and optionally
- (g) recovering or purifying the protein of interest from the host cell and/or the cell culture media.
A method of making recombinant proteins using one or more apoptosis inhibitors, comprising the steps of:
- (a) providing a vector comprising a gene encoding a protein of interest,
- (b) providing a host cell comprising a gene encoding an apoptosis inhibitor,
- (c) transforming or transfecting the host cell with the vector of step (a),
- (d) providing cell culture media,
- (e) culturing the transformed or transfected host cell in the cell culture media under conditions sufficient for expression of the protein of interest and the apoptosis inhibitor, and optionally
- (f) recovering or purifying the protein of interest from the host cell and/or cell culture media.
A method of making recombinant proteins using one or more apoptosis inhibitors, comprising-the steps of:
- a) providing a vector comprising a gene encoding a protein of interest,
- b) providing a host cell,
- c) transforming or transfecting the host cell with the vector of step (a),
- d) providing cell culture media,
- e) providing an apoptosis inhibitor,
- f) admixing the apoptosis inhibitor into the cell culture media,
- g) culturing the host cell in the cell culture media under conditions sufficient for expression of the protein of interest, and optionally
- h) recovering or purifying the protein of interest from the host cell and/or the cell culture media.
A method of increasing yield of a protein of interest in a cell culture, comprising the steps of:
- (a) providing a vector comprising a gene encoding an apoptosis inhibitor selected from the group consisting of caspase-9 dominant negative protein and baculovirus p35,
- (b) providing a vector comprising a gene encoding a protein of interest,
- (c) providing a host cell,
- (d) transforming or transfecting the host cell with the vector of steps (a) and (b),
- (e) providing cell culture media,
- (f) culturing the transformed or transfected host cell in the cell culture media under conditions sufficient for expression of the protein of interest and an amount of the apoptosis inhibitor which is effective in increasing yield of the protein of interest, and optionally
- (g) recovering or purifying the protein of interest from the host cell and/or the cell culture media.
A method of prolonging host cell viability in a cell culture, comprising the steps of:
- (a) providing a vector comprising a gene encoding an apoptosis inhibitor selected from the group consisting of caspase-9 dominant negative protein and baculovirus p35,
- (b) providing a vector comprising a gene encoding a protein of interest,
- (c) providing a host cell,
- (d) transforming or transfecting the host cell with the vector of steps (a) and (b),
- (e) providing cell culture media,
- (f) culturing the transformed or transfected host cell in the cell culture media under conditions sufficient for expression of the protein of interest and an amount of the apoptosis inhibitor which is effective for prolonging viability of the host cells in the cell culture, and optionally
- (g) recovering or purifying the protein of interest from the host cell and/or the cell culture media.
Dependencies are inferred from the claim text.