Distribution of engineered-cysteine caps
AssigneeSeattle Genetics, Inc.
InventorsSwapnil Bhargava, Cheng-Wei Aaron Chen, Matthew J. Leith
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Abstract
The present invention provides a method for removing cysteine caps from antibodies and re-capping the antibodies with cysteine molecules. The methods include, inter alia, culturing a host cell comprising a protein molecule having at least one capped engineered cysteine residue, and contacting the cell culture with cystine. Dissolved oxygen levels can be manipulated in the cell culture to further enhance the removal and re-capping process.
Claims23
A method for removing a cysteine cap from a protein molecule, the method comprising:
- culturing a host cell comprising a protein molecule having at least one capped cysteine residue;
- and
- contacting the host cell culture with cystine;
- whereby the cysteine cap is removed from the protein molecule.
A method for removing and re-forming a cysteine cap from a protein molecule, the method comprising:
- culturing a host cell comprising a protein molecule having at least one capped cysteine residue;
- and
- contacting the host cell culture with cystine;
- simultaneously or subsequent to contacting the host cell culture with cystine, contacting the host cell culture with a first manipulation of DO at a set point of 0%-50%;
- subsequent to contacting the host cell culture with DO at a set point of 0%-50%, contacting the host cell culture with a second manipulation of DO at a set point of 20%-100%;
- whereby the cysteine cap is removed from the protein molecule and the protein molecule is re-capped with a cysteine molecule.
A method for removing a cysteine cap from a protein molecule, the method comprising:
- culturing a host cell comprising a protein molecule having at least one capped cysteine residue;
- and
- contacting the host cell culture with a symmetrical disulfide;
- whereby the cysteine cap is removed from the protein molecule.
Dependencies are inferred from the claim text.