Methods for purification of recombinantly produced proteins
US5530100A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | May 7, 1990 |
| Grant date | Jun 25, 1996 |
| Priority date | — |
| Expiry date | May 7, 2010 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC07K14/755
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Recovery of the 52/48 kDa tryptic fragment of vWF or peptide subfragments thereof produced in the form of inclusion bodies from recombinant host cells is carried out by providing a washed recombinant host cell suspension, adding a detergent and subjecting the cells to mechanical disruption. A second detergent is added, followed by another mechanical disruption and centrifugation to a pellet. The pellet is resuspended in a buffer and subjected to another mechanical disruption. Inclusion bodies are washed, resuspended and then recovered. In addition, endotoxins and DNA are removed from inclusion bodies containing the 52/48 kDa tryptic fragment of vWF or peptide subfragments thereof by mechanically disrupting the inclusion bodies in an aqueous buffer containing a detergent. A washed pellet is formed from these mechanically disrupted inclusion bodies and dissolved in a denaturant. Alkylation is then performed and the suspension is subjected to column chromatography, where the alkylated fragment or subfragment is recovered by elution.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.