Purification of a pertussis outer membrane protein
US6444211B1 · kind B1 · utility
Assignee
Inventors
Key dates
| Filing date | Jun 8, 1999 |
| Grant date | Sep 3, 2002 |
| Priority date | — |
| Expiry date | Jun 8, 2019 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY10S530/825
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Pertactin (formerly 69 kDa protein) is recovered in stable biologically pure form having no detectable adenylate cyclase activity from fermentation broth from the fermentation of Bordetella pertussis as well as from the cells. The broth is processed to selectively remove pertussis toxin (PT) and filamentous haemagglutinin (FHA), the pertactin is precipitated by ammonium sulphate and the precipitate is dissolved in buffer at pH 6.0 to 8.5, the solution then is passed through hydroxyapatite and ion-exchange chromatograph columns before final ultrafiltration. Cells are extracted with urea and the extract ultrafiltered and diafiltered. The pertactin is precipitated from the extract and the precipitate processed as above. In a variation, the broth is contacted with ammonium sulphate to precipitate pertactin, PT and FHA, the precipitate is dissolved and the PT and FHA selectively removed, before the solution is passed to the chromatograph columns.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.