Claims
- 1. A method for purifying a biologically active agent comprising a factor isolated from Type 1 human female breast cyst fluid, said factor being immunoreactive with digoxin antibodies and having anti-hypertensive activity in mammals, said method comprising the steps of:
- agitating Type 1 human female breast cyst fluid with a lower alcohol, thereby causing precipitation of undesirable constituents in said fluid,
- discarding the precipitate,
- mixing the alcohol-solubilized factor with first polar solvent to form a second precipitate,
- discarding the second precipitate,
- evaporating said first polar solvent and recovering a first aqueous phase containing said factor,
- further extracting said first aqueous phase with a second polar solvent, said second polar solvent being less polar than said first polar solvent
- obtaining an organic phase containing said agent,
- removing said second polar solvent from said organic phase and obtaining a second aqueous phase containing said agent,
- lyophilizing said second aqueous phase,
- extracting said lyophilizate with methanol-weater,
- subjecting said extract to chromatography on pressure stabilized polydextran, and recovering fractions containing said agent.
- 2. The method of claim 1 wherein said agent is purified by chromatography on Sephadex LH-20.
- 3. A method for purifying a biologically active agent comprising a factor isolated from Type 1 human female breast cyst fluid, said factor being immunoreactive with digoxin antibodies and having anti-hypertensive activity in mammals, said method comprising the steps of:
- agitating Type 1 human female breast cyst fluid with a first polar solvent, thereby causing a precipitation of undersirable constituents in said fluid,
- discarding the precipitate,
- mixing the solubilized factor with a second polar solvent, and recovering an organic phase containing said factor, said second polar solvent being less polar than said first polar solvent,
- removing said second polar solvent from said organic phase and obtaining an aqueous phase containing said agent,
- lyophilizing said aqueous phase,
- extracting said lyophilizate with a methanol-water mixture,
- subjecting said extracted lyophilizate to hydrophobic affinity chromatography, and
- recovering fractions containing said agent.
- 4. The method of claim 2 wherein said agent is further purified by chromatography on an amino carbohydrate column.
- 5. The method of claim 2 wherein said agent also has the property of stimulating the binding of ouabain to the cellular ouabain receptor.
- 6. The method of claim 2 wherein said agent is further characterized by absorbing light at 210 nm of the spectrum.
FIELD OF THE INVENTION
This application is a continuation-in-part of U.S. patent application Ser. No. 939,552, filed Dec. 9, 1986 of Fred I. Chasalow and H. Leon Bradlow.
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Continuation in Parts (1)
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Number |
Date |
Country |
Parent |
939552 |
Dec 1986 |
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