Process for preparing high purity protein A preparation
AssigneeRepligen Corporation
InventorsRichard N. Love, Albert T. Profy
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Abstract
The invention concerns a process for purifying protein A preparations to high purity with high product yield. Where the protein A is obtained from a Gram-negative recombinant microbe hosting a vehicle containing a gene encoding protein A, the protein A is purified to high purity, and, advantageously, to very low levels of endotoxin. The protein A preparations made via the invention process are useful in therapeutic application, e.g., therapeutic plasma exchange, as well as for other well-known uses of protein A.
Claims42
A process for purifying protein A from a protein A-containing solution or other protein A preparation which comprises
- (a) heating a protein A-containing solution at a temperature of about 50.degree. C. to about 80.degree. C. for about 30 minutes, causing some contaminating protein to precipitate;
- (b) loading an ion-exchange column by passing the protein A-containing solution over said column;
- (c) washing the loaded column with a solution comprising a non-ionic detergent;
- (d) eluting protein A from the ion-exchange column;
- and
- (e) precipitating the eluted protein A with ethanol to give a substantially pure preparation of protein A.
A process for purifying protein A from a protein A-containing solution or other protein A preparation which comprises
- (a) heating a protein A-containing solution at a temperature of about 50.degree. C. to about 80.degree. C. for about 30 minutes, causing some contaminating proteins to precipitate;
- (b) precipitating the protein A with ethanol and re-dissolving the pellet in a buffer;
- (c) loading an ion-exchange column by passing the re-dissolved protein A-containing solution over said column;
- (d) washing the loaded column with a solution comprising a non-ionic detergent;
- (e) eluting protein A from the ion-exchange column;
- and
- (f) precipitating the eluted protein A with ethanol to give a substantially pure preparation of protein A.
Dependencies are inferred from the claim text.