Isolation of human interferon by immunosorbent and high performance liquid chromatography
AssigneeCetus Corporation
InventorsYin H. Tan, Heidi Smith-Johannsen
Life of this patent
Estimating the term…
Abstract
An improved process for the isolation and purification of HIFNs is disclosed wherein a partially purified preparation of the HIFN is sequentially passed through an antibody affinity column and a reversed-phase high performance liquid chromatographic column. Organic solvents used during the elution are extracted and the protein concentrated for subsequent use.
Claims12
A process for the isolation and purification of a lipophilic interferon protein comprising
- (a) applying a solution of said protein to an immunosorbent adsorption affinity column;
- (b) eluting said protein from said affinity column with an equilibrating buffer;
- (c) passing the eluate from said affinity column containing said protein through a reversed-phase high performance liquid chromatographic column;
- and
- (d) eluting said protein from said chromatographic column with an organic solvent.
A process for the isolation and purification of biologically active human interferon comprising:
- (a) applying a solution of ammonium sulfate precipitated HIFN to an antibody affinity column;
- (b) eluting said HIFN from said affinity column with a buffer;
- (c) ultrafiltering the eluate;
- (d) injecting said ultrafiltered HIFN solution onto a reverse-phase high performance liquid chromatographic column;
- (e) eluting said HIFN from said chromatographic column with a linear gradient flow of n-propanol;
- (f) extracting said n-propanol from the eluate with cyclohexane;
- and
- (g) remvoing residual cyclohexane from said HIFN solution with a stream of nitrogen.
A process for preparing human fibroblast interferon which process comprises
- (a) applying a solution at neutral pH of the human fibroblast interferon to an immunosorbent affinity column comprising antibodies to human fibroblast interferon immobilized on a solid support;
- (b) eluting the human fibroblast interferon with a buffer of pH about 2-3;
- (c) applying the human fibroblast interferon containing fractions from (b) to a reverse phase, high performance liquid chromatography column;
- (d) eluting the human fibroblast interferon from said chromatography column with a linear gradient flow of n-propanol.
Dependencies are inferred from the claim text.