Purification directed cloning of peptides using carbonic anhydrase as the affinity binding segment

Patent 5595887 Issued on January 21, 1997. Estimated Expiration Date:

January 21, 2014. Estimated Expiration Date is calculated based on simple USPTO term provisions. It does not account for terminal disclaimers, term adjustments, failure to pay maintenance fees, or other factors which might affect the term of a patent.

Abstract Claims Description Full Text

Patent References

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Inventors

Application

No. 552810 filed on 07/16/1990

US Classes:

435/69.7, Fusion proteins or polypeptides435/68.1Enzymatic production of a protein or polypeptide (e.g., enzymatic hydrolysis, etc.)

Examiners

Primary: Wityshyn, Michael G.
Assistant: Weber, Jon P.

Attorney, Agent or Firm

Merchant & Gould, Smith, Edell, Welter & Schmidt

Foreign Patent References

0131363 EP. 01/19/1985
0163406 EP. 12/19/1985
0230869 EP. 08/19/1987
0244147 EP. 11/19/1987
0286239 EP. 10/19/1988
0327522 EP. 08/19/1989
0333691 EP. 09/19/1989
3523701 DE. 01/19/1987
62-283998 JP. 12/19/1987
2162190 GB. 01/19/1986
2180539 GB. 04/19/1987
WO86/02077 WO. 04/19/1986
8807085 WO. 09/19/1988

International Classes

C12P 021/04
C12P 021/06

Abstract

Methods are presented for producing and purifying a variable fusion polypeptide which can be purified by affinity chromatography with the binding protein partner. The variable fusion polypeptide construct has tandem coupled segments containing one or more copies of a desired peptide linked to carbonic anhydrase as the purification binding protein. In the methods, the fusion protein is expressed in a recombinant host using a recombinant vector containing a gene encoding the fusion polypeptide. Then the expressed fusion polypeptide is purified by immobilized reversible inhibitor affinity chromatography. Finally, the purified fusion polypeptide is cleaved from the desired peptides by chemical or enzymatic means and the desired peptides purified with affinity chromatography.

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