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US 5,391,713 AGranted
Interferon purification process
AssigneeBionative AB
InventorH.ang.kan Borg
Life of this patent
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Abstract
A process for purification of human leukocyte interferon is described. The process includes immunoaffinity chromatography, ion-exchange chromatography, and a series of precipitation and centrifugation steps.
Claims25
A process for the purification of a crude human leukocyte interferon comprising the steps:
- a) applying a solution of said crude interferon onto an immuno-affinity adsorption column;
- b) eluting the adsorbed interferon from said column using a buffer solution of acid pH;
- c) concentrating the eluate resulting from step b) with a thiocyanate solution by either precipitation or ion exchange chromatography;
- d) precipitating the interferon present in the concentrated eluate resulting from step c) with a thiocyanate solution;
- e) reprecipitating the interferon resulting from step d) in aqueous ethanol;
- and
- f) recovering the interferon obtained in step e).
4 more elements
A process for purification of a crude human leukocyte interferon composition, comprising the steps:
- a) applying a crude human leukocyte interferon composition to an immunoaffinity column comprising anti-human leukocyte interferon antibodies;
- b) washing the column with a buffer;
- c) desorbing the human leukocyte interferon from the immunoaffinity column using an acidic buffer solution at a pH of about 2.0;
- d) applying the resultant desorbed human leukocyte interferon containing composition to an ion exchange column;
- e) eluting the human leukocyte interferon from the ion exchange column by increasing the pH through the addition of Tris-buffer until the resultant pH is about 8;
- f) precipitating interferon from the resultant eluate by the addition of a potassium thiocyanate solution;
- g) reprecipating interferon from the resultant sodium thiocyanate precipitate using acidic ethanol having a temperature of about -20.degree. C.;
- h) adding to the resultant supernatant a basic solution until the resultant pH is about 8.0;
- i) centrifuging the resultant precipitate and dissolving in a sodium phosphate buffer/KSCN solution;
- and
- j) subjecting the resultant interferon solution to centrifugation followed by gel filtration on a column to recover a product which exhibits an absorbance at 280 nm.
8 more elements
Dependencies are inferred from the claim text.