Polysaccharide toxin from Group B .beta.-hemolytic Streptococcus (GBS) having improved purity

• US 5,811,403 A
• Filed: 09/30/1996
• Issued: 09/22/1998
• Est. Priority Date: 09/30/1996
• Status: Expired due to Term

• Current Assignee: Vanderbilt University
• Original Assignee: Vanderbilt University
• Inventors: Hellerqvist, Carl G.
• Primary Examiner(s): Weber, Jon P.
• Assistant Examiner(s): HANLEY, SUSAN MARIE
• Application Number:
  US08/744,770
• Time in Patent Office: 722 Days
• Field of Search: 435/252.1, 435/253.4, 435/72, 435/73, 514/25, 514/23, 514/42, 514/43, 514/54, 514/57, 514/60, 530/414, 530/416, 530/415, 536/6
• US Class Current: 514/23
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 1/205 Bacterial isolates
• CPC Class Codes: C12P 1/04 by using bacteria
• CPC Class Codes: C12P 19/26 Preparation of nitrogen-containing carbohydrates
• CPC Class Codes: C12R 2001/46 Streptococcus ; Enterococcus; Lactococcus

Abstract

The invention relates to a polysaccharide toxin from group B β-hemolytic Streptococcus (GBS) having improved purity. The improved purity of the toxin, 95%, is achieved by the method of purification wherein a bacterial fermentation stock is subjected to chromatography with hydrophobic interaction chromatography (HIC) resin, extraction with a phenol/saline solution followed by ion exchange chromatography. The purified GBS toxin has a molecular weight of about 300 kD and a relative carbohydrate ratio of rhamnose:mannose:galactose:glucose:N-acetyl glucosamine:N-acetyl galactosamine of about 0:1:3:1:1:1, respectively. The GBS toxin is useful for treatment of tumors and other medical conditions.

Claims

1. A group B β

-hemolytic Streptococcus (GBS) polysaccharide toxin having at least about 95% purity, a molecular weight of about 300 kD and a relative carbohydrate ratio of rhamnose;

mannose;

galactose;

glucose;

N-acetyl glucosamine;

N-acetyl galactosamine of about 0;

1;

3;

1;

1;

1, respectively.

3. A pharmaceutical composition comprising the GBS toxin of claims 1 or 2 and a pharmaceutically acceptable carrier.

2. A group B β

-hemolytic Streptococcus (GBS) polysaccharide toxin having at least about 95% purity obtained by the steps comprising;

a) applying a source of said GBS toxin to a hydrophobic interaction chromatography (HIC) resin;

b) eluting the HIC resin to obtain an eluate containing said GBS toxin;

c) extracting said GBS toxin-containing eluate with an aqueous salt solution of phenol;

d) recovering said GBS toxin from the extraction step;

e) applying said GBS toxin recovered from the extraction step to a column having ion exchange resin;

f) eluting the ion exchange column and recovering said GBS toxin from the ion exchange eluate; and

wherein said recovered GBS toxin has a molecular weight of about 300 kD and a relative carbohydrate ratio of rhamnose;

mannose;

galactose;

glucose;

N-acetyl glucosamine;

N-acetyl galactosamine of about 0;

1;

3;

1;

1;

1, respectively.

4. The GBS toxin according to claim 2 wherein the HIC resin has interactive hydrophobic groups bound to said resin and wherein said hydrophobic groups are selected from the group consisting of alkyl, alkoxy and aryl.

5. The GBS toxin according to claim 4 wherein said hydrophobic groups are alkyl groups consisting of 2 to 12 carbons.