Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Separation or purification
Patent
1994-07-15
1997-03-11
Ulm, John
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Separation or purification
530416, 530417, 530422, 530425, 530351, 530426, 435 6952, 435 711, C07K 114, C12N 1519
Patent
active
056102844
ABSTRACT:
There are disclosed (i) a purification process for obtaining a human BCDF having the intramolecular disulfide linkage and the stereostructure of natural type human BCDF which comprises subjecting to an oxidation reaction and a refolding treatment a reduced type human BCDF obtained by culturing a microorganism having a human BCDF gene integrated therein and solubilized with guanidine hydrochloride, characterized in that after the oxidation reaction, a gel filtration chromatographic treatment is conducted under the conditions of the guanidine hydrochloride concentration adjusted to 4-7M; (ii) a purification process for obtaining a natural type human BCDF monomer by removing the organic solvent from an organic solvent-containing solution of human BCDF, characterized in that the solution is passed through a gel filtration chromatographic column equilibrated with an organic solvent, followed by eluting according to a stepwise or linear gradient program; and (iii) a human BCDF purification process comprising an ion exchange chromatographic treatment and a reversed phase HPLC step, in combination (i) or (ii). According to these purification processes, it becomes possible to remove the impurities derived from a microorganism and human BCDF analogs, and the thus obtained natural type human BCDF has a high purity and can be utilized for pharmaceutical preparations.
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Date Masayo
Ejima Daisuke
Sato Yutaka
Takahara Yoshiyuki
Watanabe Mayumi
Ajinomoto Co. Inc.
Mertz Prema
Ulm John
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