Method of separating glycosylated and non-glycosylated prolactin

Details Method of separating glycosylated and non-glycosylated prolactin Method of separating glycosylated and non-glycosylated prolactin

1993-05-04

1994-09-06

Naff, David M.

Chemistry: natural resins or derivatives; peptides or proteins;

Proteins, i.e., more than 100 amino acid residues

Separation or purification

530399, A23J 100, A61K 3724

Patent

active

053449207

ABSTRACT:
This invention relates to a method for separating glycosylated prolactin from non-glycosylated prolactin. The method comprises the following steps: 1) equilibrating the prolactin containing solution with an equilibrating solution containing at least one organic solvent; 2) adding the equilibrated prolactin solution onto a chromatographic column containing a packing for the ion-exchange affinity separation of glycosylated and non-glycosylated prolactin; 3) eluting the glycosylated prolactin from the chromatographic column with a sufficient quantity of a first eluting solution containing at least one organic solvent and with or without at least one salt, wherein the salt concentration and/or organic solvent concentration and/or pH are appropriate for the eluting of glycosylated prolactin being substantially free of non-glycosylated prolactin; and 4) eluting the non-glycosylated prolactin from the chromatographic column with a sufficient quantity of a first eluting solution containing at least one organic solvent and with or without at least one salt, wherein the salt concentration and/or organic solvent concentration and/or pH are appropriate for the eluting of non-glycosylated prolactin being substantially free of glycosylated prolactin.

REFERENCES:
Kacsoh et al., Endocrine Regulations, vol. 25, 98-110, 1991.
Lewis et al., Endocrinology, vol. 116, No. 1, pp. 359-363, 1985.
Young et al., Mol. & Cellular Endocrinology, 71 (155-162), 1990.
Chapitis et al., Am. J. Obstet. Gynecol., Apr. 1988, vol. 158, No. 4, pp. 846-853.
Pharmacia Fine Chemicals Booklet, Mar. 1980, pp. 32-38.

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