Method of separating glycosylated and non-glycosylated prolactin by ion exchange chromatography

A method for separating glycosylated prolactin from non-glycosylated prolactin comprising:

• a. equilibrating a solution containing glycosylated and non-glycosylated prolactin with an equilibrating solution containing at least one lower alcohol and/or lower glycol;
• b. adding the equilibrated prolactin solution onto a chromatographic column containing a cationic or anionic packing for the ion exchange separation of glycosylated and non-glycosylated prolactin;
• c. eluting the glycosylated prolactin from the chromatographic column with a sufficient quantity of a first eluting solution comprising:
• i. at least one lower alcohol and/or lower glycol in concentration less than 50% per total volume;
• and
• ii. at least one salt, wherein the salt concentration, lower alcohol and/or lower glycol concentration and pH are appropriate for the eluting of non-glycosylated prolactin being substantially free of glycosylated prolactin;
• and
• d. eluting the glycosylated prolactin from the chromatographic column with a sufficient quantity of a second eluting solution comprising:
• i. at least one lower alcohol and/or lower glycol in concentration less than 50% per total volume;
• and
• ii. at least one salt,
• wherein the salt concentration, lower alcohol and/or lower glycol concentration and pH are appropriate for the eluting of glycosylated prolactin being substantially free of non-glycosylated prolactin.

A method for separating glycosylated prolactin from non-glycosylated prolactin comprising:

• a. equilibrating a solution containing glycosylated and non-glycosylated prolactin with an equilibrating solution containing at least one lower alcohol and/or lower glycol;
• b. adding the equilibrated prolactin solution onto a chromatographic column containing a cationic or anionic packing for the ion exchange separation of glycosylated and non-glycosylated prolactin;
• c. eluting the glycosylated prolactin from the chromatographic column with a sufficient quantity of a first eluting solution containing at least one lower alcohol and/or lower glycol in concentration less than 50% per total volume;
• wherein the lower alcohol and/or lower glycol concentration and pH are appropriate for the eluting of glycosylated prolactin being substantially free of non-glycosylated prolactin;
• and
• d. eluting the non-glycosylated prolactin from the chromatographic column with a sufficient quantity of a second eluting solution comprising:
• i. at least one lower alcohol and/or lower glycol in concentration less than 50% per total volume;
• and
• ii. at least one salt,
• wherein the salt concentration, lower alcohol and/or lower glycol concentration and pH are appropriate for the eluting of non-glycosylated prolactin being substantially free of glycosylated prolactin.

A method for separating glycosylated prolactin from non-glycosylated prolactin comprising:

• a. equilibrating a solution containing glycosylated and non-glycosylated prolactin with an equilibrating solution containing at least one lower alcohol or lower glycol;
• b. adding the equilibrated prolactin solution onto a chromatographic column containing a cationic or anionic packing for the ion exchange separation of glycosylated and non-glycosylated prolactin;
• c. eluting the glycosylated prolactin from the chromatographic column with a sufficient quantity of a first eluting solution comprising:
• i. at least one lower alcohol and/or lower glycol in concentration less than 50% per total volume;
• and
• ii. at least one salt, wherein the salt concentration, lower alcohol and/or lower glycol concentration and pH are appropriate for the eluting of glycosylated prolactin being substantially free of non-glycosylated prolactin;
• and
• d. eluting the non-glycosylated prolactin from the chromatographic column with a sufficient quantity of a second eluting solution containing at least one lower alcohol and/or lower glycol in concentration less than 50% per total volume, wherein the lower alcohol and/or lower glycol concentration and pH are appropriate for the eluting of non-glycosylated prolactin being substantially free of glycosylated prolactin.

A method for separating glycosylated prolactin from non-glycosylated prolactin comprising:

• a. equilibrating a solution containing glycosylated and non-glycosylated prolactin with an equilibrating solution containing at least one lower alcohol and/or lower glycol;
• b. adding the equilibrated prolactin solution onto a chromatographic column containing a cationic or anionic packing for the ion exchange separation of glycosylated and non-glycosylated prolactin;
• c. eluting the glycosylated prolactin from the chromatographic column with a sufficient quantity of a first eluting solution containing at least one lower alcohol and/or lower glycol in concentration less than 50% per total volume, wherein the lower alcohol and/or lower glycol concentration and pH are appropriate for the eluting of glycosylated prolactin being substantially free of non-glycosylated prolactin;
• and
• d. eluting the non-glycosylated prolactin from the chromatographic column with a sufficient quantity of a second eluting solution containing at least one lower alcohol and/or lower glycol in concentration less than 50% per total volume, wherein the lower alcohol and/or lower glycol concentration and pH are appropriate for the eluting of non-glycosylated prolactin being substantially free of glycosylated prolactin.