Affinity peptides

• US 5,284,933 A
• Filed: 02/22/1988
• Issued: 02/08/1994
• Est. Priority Date: 03/10/1987
• Status: Expired due to Term

• Current Assignee: Roche Diagnostic Systems Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: Eggimann, Bernhard, Gentz, Reiner, Hochuli, Erich, Stuber, Dietrich, Dobeli, Heinz
• Primary Examiner(s): Ossanna, Nina
• Application Number:
  US07/158,962
• Time in Patent Office: 2,178 Days
• Field of Search: 530/350, 530/324, 530/413, 530/409, 435/69.7
• US Class Current: 530/350
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07K 14/57 IFN-gamma
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/21 containing a His-tag
• CPC Class Codes: C07K 2319/50 containing protease site
• CPC Class Codes: C07K 2319/75 containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/70 Vectors or expression systems specially adapted for E. coli

Abstract

Fusion proteins and a process for their purification by means of metal chelate affinity chromatography on NTA resins are provided by this invention.

Claims

1. A fusion protein comprising a biologically active polypeptide or protein linked by its amino- and/or carboxyl-terminus to one or two affinity peptides, which affinity peptides have the formula

View Dependent Claims (2, 3, 4, 5, 6, 7, 8)

2. The fusion protein of claim 1 in which the affinity peptides have an amino acid sequence selected from the group consisting of ##STR3##

3. The fusion protein of claim 1 in which at least one of the links between the biologically active polypeptide or protein and the affinity peptides can be cleaved by a protease.

4. The fusion protein of claim 3 in which the protease is enterokinase.

5. The fusion protein of claim 3 in which the protease is coagulation factor X_a.

6. The fusion protein of claim 1 in which the affinity peptides complex immobilized nickel ions.

7. The fusion protein of claim 1 in which the biologically active polypeptide or protein has an amino acid sequence corresponding to the sequence of human immune interferon, a subsequence of human immune interferon, or mouse dihydrofolate reductase.

8. The fusion protein of claim 1 which is essentially pure.

9. An affinity peptide of formula

View Dependent Claims (10)

10. The affinity peptide of claim 9 having an amino acid sequence selected from the group consisting of ##STR4##