Methods and compositions; purified preparation of neural progenitor regulatory factor

• US 5,276,145 A
• Filed: 03/17/1992
• Issued: 01/04/1994
• Est. Priority Date: 08/04/1989
• Status: Expired due to Term

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Bottenstein, Jane E.
• Primary Examiner(s): Russel, Jeffrey E.
• Application Number:
  US07/852,755
• Time in Patent Office: 658 Days
• Field of Search: 530/350, 530/399, 514/12, 514/21, 514/2, 514/8
• US Class Current: 530/399
• CPC Class Codes: C07K 14/475 Growth factors; Growth regulators

Abstract

A novel substantially purified preparation of a neural progenitor regulatory factor and methods for producing such purified factor are claimed. In a preferred embodiment, the factor has an approximate molecular weight of about 46-47 kilodaltons (as de

FUNDING

Development of the present invention was facilitated by funding from the National Institutes of Health, Grant # NS 20375. Accordingly, the U.S. Government may own certain rights.

Claims

1. A purified preparation, which is substantially free of platelet derived growth factor, of a neural progenitor regulatory factor, wherein said factor has:

a) a molecular weight between about 40 and about 50 kilodaltons as determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis; and

b) an ability to stimulate proliferation of 0-2A glial progenitor cells.

4. The composition of claims 1, 2 or 3 wherein said molecular weight is about 46-47 kilodaltons.

2. A purified preparation, which is substantially free of platelet derived growth factor, of a neural progenitor cell regulatory factor, wherein said factor has the following characteristics:

a) affinity for heparin

i) such that said factor binds to a heparin sepharose gel particle when said factor and a preparation of heparin sepharose gel particles are mixed together with a buffer comprising about 5 mM phosphate, 50 mM NaCl, and having a pH of about 7.4 for a period of about 18 hours; and

ii) elutes from said heparin sepharose particles when the concentration of NaCl in said buffer is raised above about 0.7M-0.8M;

b) a molecular weight between about 40 and about 50 kilodaltons as determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis; and

c) an ability to stimulate proliferation of cells of the C62C glial cell line.

3. A purified preparation, which is substantially free of platelet derived growth factor, of a neural progenitor cell regulatory factor, wherein said factor has the following characteristics:

a) affinity for heparin

i) such that said factor binds to a heparin sepharose gel particle when said factor and a preparation of heparin sepharose gel particles are mixed together with a buffer comprising 5mM phosphate, 50mM NaCl, and having a pH of about 7.4 for a period of about eight hours; and

ii) elutes from said heparin sepharose particles when the concentration of NaCl in said buffer is raised above about 0.7M-0.8M;

b) a molecular weight between about 40 and about 50 kilodaltons as determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis; and

c) an ability to stimulate proliferation of 0-2A glial progenitor cells derived from the brain of a neonatal rat.

5. A purified preparation, which is substantially free of platelet derived growth factor, of a neural progenitor regulatory factor wherein said factor is the neural progenitor regulatory factor produced by a cell line ATCC (American Type Culture Collection) accession number CLR 10187 and has a molecular weight between about 40 and about 50 kilodaltons as determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis.