Patent EP0104356A1 successfully refreshed from Google Patents.
← Back to Patent Explorer
EP0104356A1 Expired / Lapsed EP ⏱️ 0.0 years remaining

Highly purified human antihemophilic factor and method for the preparation thereof

Assignee: University of Rochester
•
Filing: 1983-07-28
•
Granted: —
•
Expires: 2003-07-28
📄 Original PDF 🌐 Google Patents 🔄 Sync Now

Abstract

Abstract Highly purified, biologically active Human Antihemophilic Factor (AHF) preparations are prepared having specific activities of about 4000 - 8000 units per milligram of AHF. In the method of preparation an AHF concentrate, prepared by fractionation of plasma to partially remove fibrinogen, fibronectin and other plasma components is subjected to a separation on the basis of Stokes' radius to separate AHF from the bulk of remaining proteins in the AHF concentrate. The pooled fractions containing AHF activity are concentrated by precipitation with ammonium sulfate, sodium sulfate, etc., by diafiltration, by PEG addition, or the like. The concentrate, is solubilized or equilibrated in an aqueous medium and treated to change the effective Stokes' radius of the AHF to an apparently low value and then subjected to a separation from the concentrate. The AHF pool from above is treated to remove cations by dialysis against an appropriate buffer of lower ionic strength and chromatographed on an anion-exchange medium. The AHF fraction from the above chromatography, is a highly purified AHF preparation.

Inventors

  • 👤 Stephen I. Chavin
  • 👤 Philip J. Fay

Bibliographic Details

Application Number —
Jurisdiction EP
Data Source google_patents
Last Synchronized 2026-11-02 22:53
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/745; C07K14/755; A; A61; A61K; A61K38/00; Y; Y10; Y10S; Y10S530/00; Y10S530/827; Y10S530/829; Y10S530/83

Claims Analysis

Total Claims: 10
Claim 1
Independent Claim 132 words
l. A method for preparing a highly purified. essentially homogenous Antihemophilic Factor concentrate,characterized in that it comprises the steps of - (a) obtaining an Antihemophilic Factor concentrate which is totally or partially free from prothrombin complex proteins, fibrinogen, and albumin, (b) subjecting the Antihemophilic Factor concentrate to a separation on the basis of Stoke's radius to separate Antihemophilic Factor of an apparently high Stoke's radius value from other proteins, (c) treating the Antihemophilic Factor concentrate to change the effective Stoke's radius of the Antihemophilic Factor molecule to an apparently low value, (d) subjecting the Antihemophilic Factor concentrate to a separation on the basis of Stoke's radius to separate Antihemophilic Factor of apparently low Stoke's radius value from other proteins, (e) subjecting the Antihemophilic Factor concentrate to chromotography on an anion exchange medium.
Claim 2
Dependent Claim 35 words
A method according to claim 1, characterized in that the separation in step (b) or step (d) is accomplished by subjecting the Antihemophilic Factor concentrate to gel permeation chromatography on cross-linked agarose or cross-linked polyacrylamide.
Claim 3
Dependent Claim 31 words
A method according to claim 1, characterized in that the separation in step (b) and/or step (d) is accomplished by subjecting the Antihemophilic Factor concentrate to controlled pore glass bead chromatography.
Claim 4
Dependent Claim 24 words
A method according to claim 1, characterized in that the separation in step (b) and/or step (d) is accomplished by sucrose density gradient ultrafiltration.
Claim 5
Independent Claim 55 words
A method according to any of claims 1 to 4, characterized in that the Antihemophilic Factor concentrate in step (c) is treated. with a source of divalent cations in an amount of about 5 - 10 parts by volume of Antihemophilic concentrate per part of a solution about 1 - 3 M in divalent cations.
Claim 6
Dependent Claim 19 words
A highly purified, essentially homogenous Antihemophilic Factor concentrate obtainable by the method of any of claims 1 to 5.
Claim 7
Dependent Claim 20 words
An Antihemophilic Factor concentrate having a specific activity of at least about 4000 Antihemophilic Factor units per milligram of protein.
Claim 8
Dependent Claim 29 words
An Antihemophilic Factor concentrate according to claim 7, characterized in that it is essentially free from Factors II, VII, IX, and X, fibriongen, albumin, fibronectin, and von Willebrand factor.
Claim 9
Dependent Claim 30 words
An Antihemophilic Factor concentrate according to any of claims 6 to 8, characterized in that it has the amino acid composition of the Antihemophilic Factor as given in Table 1.
Claim 10
Dependent Claim 30 words
An Antihemophilic Factor concentrate according to any of claims 6 to 9, characterized in that the ratio of Antihemophilic Factor to Antihemophilic Factor antigen is about 50: 1 or greater.

References Cited (Backward Citations)

US4278594A Cited
EP0041173A1 Cited

USPTO Annuity & Maintenance Windows

Maintenance schedule is calculated for granted US utility patents.

Assigned Categories & Tags

No categories assigned to this patent yet.

Assign Category Tag