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US 4,920,097 AGranted
Immunosuppressor and method of extraction thereof from leukocytes
AssigneeImreg, Inc.
InventorsA. Arthur Gottlieb, Robert C. Sizemore, Sudhir K. Sinha
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Abstract
Methods of extracting an immunosuppressor from leukocyte dialysates are disclosed, together with methods for using the immunosuppressor and compositions containing the immunosuppressor.
Claims20
In a method of extracting a suppressor from an extract of leukocytes and purifying it whereby said suppressor is separated from extraneous materials in said extract, comprising the
- (1) size fractionating said extract, thereby producing a first dialysate having a nominal M.W. cutoff of approximately 12,000;
- (2) size fractionating said first dialysate, thereby producing a seond dialysate and a retentate, separated from one another at a nominal M.W. cutoff of approximately 3500;
- (3) applying said retentate to a reverse phase octadecylsilane HPLC column;
- (4) eluting said column with a solvent system;
- the improvement comprising:
- eluting said column with an acetonitrile in dilute aqueous trifluoroacetic acid gradient, said gradient including a range of acetonitrile concentration from approximately 70% to approximately 80%, thereby producing elutants associated with different portions of said gradient;
- and collecting the elutant associated with the zone of said gradient wherein acetonitrile concentration is between approximately 75% and approximately 80%, whereby material designated herein as L-4 suppressor is provided.
4 more elements
A material having a M.W. between 3500 and 12,000, and further characterized by having the following properties:
- (a) being substantially entirely chromotographically elutable from octadecylsilane by acetonitrile in dilute aqueous trifluoroacetic acid gradient, in the portions of said gradient between approximately 70% and approximately 85% acetonitrile concentration;
- (b) being substantially not so elutable in the portions of said gradient below approximately 70% acetonitrile concentration;
- and
- (c) being substantially free of material that is so elutable below said concentration and of material that is so elutable only above 90% acetonitrile concentration;
- (d) not being chromotographically elutable from hydroxylapatite by aqueous ammonium bicarbonate gradient, in the portions of said gradient between approximately 0.1 M and 0.15 M;
- and
- (e) nonspecifically causing a lessening of the magnitude or rate of an immune system reaction when administered to a human or other mammalian subject.
4 more elements
Dependencies are inferred from the claim text.