Methods for harvesting biomolecules
AssigneeAmgen Inc.
InventorsThomas Clark Pearson, Sarah Whetstone, Jeremy S. Conner
Life of this patent
Estimating the term…
Abstract
The disclosure provides methods of purifying native and recombinant biomolecules, e.g., proteins, from mammalian cells using purification protocols incorporating harvest recovery operations involving decanter centrifugation of at least one target biomolecule from at least one particulate component of cell culture fluid. The unexpected capacity of decanter centrifuge separation of biological materials of similar densities found in mammalian cell culture fluid has been found to yield high quantities of functional protein in efficient, low-cost harvest recovery steps of biomolecule purification protocols.
Claims17
A method for separating protein produced in a mammalian cell from at least one other mammalian cell component comprising:
- (a) introducing a continuous stream of a fluid comprising mammalian cells from a bioreactor into at least one decanter centrifuge;
- (b) operating the decanter centrifuge, thereby separating fluid components into a low density composition and a high density composition;
- (c) recycling the low density composition through the bioreactor;
- (d) collecting the low density composition or recycling the low density composition through the decanter centrifuge;
- and
- (e) recycling the high density composition through the decanter centrifuge,
- wherein the fluid components comprise mammalian cells and components of mammalian cells, further wherein the low density composition comprises the protein.
A method for purifying a protein from a mammalian cell culture comprising:
- (a) introducing a continuous stream of a cell culture fluid comprising mammalian cells into at least one decanter centrifuge;
- (b) operating the decanter centrifuge, thereby separating fluid components into a low density fluid composition and a high density fluid composition;
- (c) subjecting the low density fluid composition to depth filtration to yield a filtered low density fluid composition;
- (d) recycling the filtered low density composition through the bioreactor to obtain a recycled low density composition;
- (e) recycling the high density composition through the decanter centrifuge;
- (f) contacting the recycled low density fluid composition with at least one chromatography medium;
- and
- (g) collecting fluid containing the protein from the chromatography medium, thereby purifying the protein.
Dependencies are inferred from the claim text.