Isolation of human plasma procoagulant protein factor VIII from biological factors
US4675385A · kind A · utility
Assignee
Inventor
Key dates
| Filing date | Mar 27, 1985 |
| Grant date | Jun 23, 1987 |
| Priority date | — |
| Expiry date | Mar 27, 2005 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY10S530/83
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
A rapid and simple process for purifying human, bovine and porcine procoagulant protein Factor VIII on a large scale using sequential high performance size exclusion chromatography under, first, low salt concentration conditions and, second, under high salt concentration conditions from reconstituted commercial Factor VIII:C (complexed Factor VIII) concentrate. The chromatographic separation is carried out on a high performance size exclusion chromatographic column packed with porous beads having a particle size of from about 13 to about 35 microns, pore diameters of from about 500 to about 2000 Angstroms and a pore volume of from about 1.0 to about 1.8 ml per gram. The first chromatographic separation is carried in a buffered aqueous solution using the buffered aqueous solution as an eluant. The low molecular weight constituents (impurities) are separated from Factor VIII and the high molecular weight constituents (impurities). A second chromatographic separation may be carried out after Factor VIII has been dissociated in a buffered solution having a concentration of from about 0.25 to about 0.45M calcium ion. The second chromatographic column is packed with some packing as the f…
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