Recombinant peptide production using a cross-linkable solubility tag

US7951559B2 · kind B2 · utility

Assignee

• E.I. DU PONT DE NEMOURS AND COMPANY · US

Inventors

• Albert W. Alsop · Wilmington, US
• Qiong Cheng · Wilmington, US
• Linda Jane Solomon · Wilmington, US
• Stephen R. Fahnestock · Wilmington, US
• Tanja Maria Gruber · Media, US
• Pierre E. Rouviere · Wilmington, US

Key dates

• Filing date: Jul 14, 2008
• Grant date: May 31, 2011
• Expiry date: Jul 14, 2029

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C07K2319/00
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The invention relates to the recombinant expression of a peptide of interest in the form of a fusion protein comprising a solubility tag. The fusion protein comprises at least two portions separated by a cleavable peptide sequence wherein one portion is devoid of cysteine residues and the second portion comprises an effective number of cross-linkable cysteine residues. After cell lysis and isolation of the fusion protein, the fusion protein is subsequently cleaved into a mixture of first and second portions. Oxidative cross-linking is used to selectively precipitate one of the two portions to facilitate simple and effective separation of the peptide of interest.