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US 7,247,475 B2Granted
Method for producing monoclonal antibodies
AssigneeWyeth
InventorsClive R. Wood, Randal J. Kaufman
Life of this patent
Estimating the term…
Abstract
An improved method for the production of monoclonal antibodies is disclosed.
Claims20
A method of optimizing the expression level of an antibody or a fragment thereof which comprises:
- (a) producing eukaryotic host cells containing and capable of expressing a first DNA sequence encoding at least an antibody heavy chain or an antigen binding portion thereof, said first DNA sequence being associated with a first heterologous selectable amplifiable marker gene, and a second DNA sequence encoding at least an antibody light chain or an antigen binding portion thereof, said second DNA sequence being associated with a second heterologous selectable amplifiable marker gene;
- (b) culturing said host cells in a suitable culture medium;
- (c) differentially amplifying said first and second DNA sequences with appropriate selective agents to allow maximized production of said antibody or fragment thereof
- (d) choosing a host cell that produces a desired amount of antibody.
1 more element
A method of optimizing expression of an antibody or fragment thereof comprising:
- (a) producing a first eukaryotic host cell containing and capable of expressing a first DNA sequence encoding at least an antibody heavy chain variable region or an antigen binding portion thereof, said first DNA sequence being associated with a heterologous selectable amplifiable marker gene;
- (b) producing a second eukaryotic host cell containing and capable of expressing a second DNA sequence encoding at least an antibody light chain variable region or an antigen binding portion thereof, said second DNA sequence being associated with a heterologous selectable amplifiable marker gene,
- (c) culturing said host cells in suitable selective culture media;
- (d) measuring the relative amounts of said first and second DNA sequences expressed;
- (e) differentially amplifying said first and said second DNA sequences with appropriate selective agent to optimize the relative gene copy number of said first and second DNA sequences;
- (f) fusing said first and second host cells;
- and
- (g) expressing said antibody or fragment thereof.
5 more elements
A method of optimizing expression of an antibody or fragment thereof comprising:
- (a) producing a first eukaryotic host cell containing and capable of expressing a first DNA sequence encoding at least an antibody heavy chain variable region or an antigen binding portion thereof, said first DNA sequence being associated with a heterologous selectable amplifiable marker gene;
- (b) producing a second eukaryotic host cell containing and capable of expressing a second DNA sequence encoding at least an antibody light chain variable region or an antigen binding portion thereof, said second DNA sequence being associated with a heterologous selectable amplifiable marker gene,
- (c) culturing said host cells in suitable selective culture media;
- (d) differentially amplifying said first and said second DNA sequences with appropriate selective agent;
- (e) fusing said first and second host cells;
- (f) choosing a host cell that produces a desired amount of antibody;
- and
- (g) expressing said antibody or fragment thereof.
5 more elements
Dependencies are inferred from the claim text.