Product and process for introduction of a hinge region into a fusion protein to facilitate cleavage
US5013653A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Dec 28, 1989 |
| Grant date | May 7, 1991 |
| Priority date | — |
| Expiry date | Dec 28, 2009 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC07K2319/75
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Disclosed is a method for the isolation and purification of polypeptides expressed in host cells by recombinant DNA techniques. A fused polypeptide is produced containing a desired polypeptide fused to additional amino acids. The additional amino acids define a leader sequence having properties exploitable in purification, a hinge region, and a cleavage site. The hinge region is cysteine-free and has a secondary structure which serves to expose the cleavage site to a selected endopeptidase. The method of the invention involves the production of a fused polypeptide which may be efficiently isolated by exploiting the properties of the leader sequence, and then efficiently cleaved at the cleavage site in an appropriate aqueous environment by virtue of the influence of the hinge on the cleavage agent/cleavage site reaction and other properties of the fused polypeptide.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.