Method for purifying heregulin
AssigneeGenentech, Inc.
InventorsRichard L. Vandlen, William E. Holmes
Life of this patent
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Abstract
A novel polypeptide with binding affinity for the p185.sup.HER2 receptor, designated heregulin-.alpha., has been identified and purified from cultured human cells. DNA sequences encoding additional heregulin polypeptides, designated heregulin-.alpha., heregulin-.beta.1, heregulin-.beta.2, heregulin-.beta.2-like, and heregulin-.beta.3, have been isolated, sequenced and expressed. Provided herein are nucleic acid sequences encoding the amino acid sequences of heregulins useful in the production of heregulins by recombinant means. Further provided are the amino acid sequences of heregulins and purification methods therefor. Heregulins and their antibodies are useful as therapeutic agents and in diagnostic methods.
Claims17
A method for increasing the purity of a composition comprising a heregulin selected from the group consisting of heregulin-.alpha. (HRG-.alpha.), heregulin-.beta.1 (HRG-.beta.1), heregulin-.beta.2 (HRG.beta.2), heregulin-.beta.2-like (HRG-.beta.2-like), and heregulin-.beta.3 (HRG-.beta.3) which has been expressed by a recombinant cell comprising the use of a procedure selected from the group consisting of fractionation on an immunoaffinity column, fractionation on an ion-exchange column, ethanol precipitation, reverse phase HPLC, chromatography on silica, chromatography on heparin Sepharose, chromatography on a cation exchange resin, chromatofocusing, SDS-PAGE, ammonium sulfate precipitation, and gel filtration, wherein the heregulin is purified to a degree of purity suitable for diagnostic use.
A method for increasing the purity of a composition comprising heregulin and a contaminant to a degree of purity suitable for diagnostic use comprising the steps of adsorbing the heregulin onto heparin Sepharose and adsorbing the heregulin onto a polyaspartic acid column, wherein the heregulin is heregulin-.alpha. (HRG-.alpha.), heregulin-.beta.1 (HRG-.beta.1), heregulin-.beta.2 (HRG-.beta.2), heregulin-.beta.2-like (HRG-.beta.2-like) or heregulin-.beta.3 (HRG-.beta.3).
Dependencies are inferred from the claim text.