# US 5,710,254 A · Purification of von Willebrand factor by affinity chromatography

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- Number: US 5,710,254 A
- Document ID: US5710254A
- Status: Granted
- Kind: A
- Application number: 08/474,623
- Filed: 1995-06-07
- Granted: 1998-01-20
- Earliest claimed: 1987-06-29
- Assignees: Rhone-Poulenc Rorer Pharmaceuticals Inc.
- Inventors: Jack Newman; David Farb
- Web page: https://patentgrep.com/patent/US5710254A

## CPC

- C07K 14/755: Factors VIII {, e.g. factor VIII C (AHF), factor VIII Ag (VWF)}
- 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)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

A method of improving the therapeutic activity of von Willebrand Factor obtained from materials comprising on Willebrand Factor comprising incubating said Factor at a temperature of about 20.degree. C. to about 55.degree. C. for about 1 to about 30 hours, most preferably at from about 45.degree. C. to about 55.degree. C. for about 1 to about 5 hours.

## Claims

### Claim 1

A method of improving the therapeutic activity of von Willebrand Factor prepared by recombinant technology, the improvement comprising: incubating a solution of said Factor, substantially free from a chaotropic agent and Factor VIII and not in contact with a column matrix solid substance, at a temperature of about 20.degree. C. to about 55.degree. C. for about 1 to about 30 hours at a pH of about 6.5 to 7.5 and with buffering, which conditions increase the activity of said Factor.

### Claim 2 (depends on 1)

The method of claim 1 wherein said Factor is incubated at a temperature of about 30.degree. C. to about 55.degree. C. for about 5 to about 15 hours.

### Claim 3 (depends on 1)

The method of claim 1 wherein said Factor is incubated in a buffer solution comprising from about 0.01 to about 0.5M Tris and from about 0.01 to about 1.5M NaCI.

### Claim 4 (depends on 3)

The method of claim 3 wherein said Factor is incubated at a temperature of about 30.degree. C. to about 55.degree. C. for about 5 to about 15 hours.

### Claim 5

A method of improving the therapeutic activity of von Willebrand Factor prepared by recombinant technology and eluted from a material comprising said Factor bound to a monoclonal or polyclonal antibody specific thereto, the improvement comprising: incubating a solution of said Factor, substantially free from a chaotropic agent and Factor VIII and not in contact with a column matrix solid substance, at a temperature of about 20.degree. C. to about 55.degree. C. for about 1 to about 30 hours at a pH of about 6.5 to 7.5 and with buffering, which conditions increase the activity of said Factor.

### Claim 6 (depends on 5)

The method of claim 5 wherein said Factor is incubated in a buffer solution comprising from about 0.01 to about 0.5M Tris and from about 0.01 to about 1.5M NaCI.