High-performance liquid chromatography of amino acids, peptides and proteins. LXXXV. Separation of isoforms of the glycoprotein hormones from human pituitary extracts.
Procedures for the high-resolution separation of microheterogeneous forms of the glycoprotein hormones, lutropin, follitropin and thyrotropin, present in human pituitary extracts are described. The sequential use of preparative isoelectric focusing and ion-exchange high-performance liquid chromatography (HPLC) allows the further discrimination of the isoforms of these glycoprotein hormones, previously considered homogeneous on the basis of their electrophoretic mobility. The ability of ion-exchange HPLC stationary phases to probe for slight differences in charge asymmetry expressed on the surface of these proteins raises the possibility of their rapid purification via the tandem on-line use of the same ion-exchange support with several different elution systems. Retention maps, illustrating the basis of this approach, are described.
D000596:Amino Acids / Q000302:isolation & purification; D002021:Buffers; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D005640:Follicle Stimulating Hormone / Q000032:analysis; D006023:Glycoproteins / Q000302:isolation & purification*; D006801:Humans; D007525:Isoelectric Focusing; D007986:Luteinizing Hormone / Q000032:analysis; D010455:Peptides / Q000302:isolation & purification; D010907:Pituitary Hormones / Q000302:isolation & purification*; D011506:Proteins / Q000302:isolation & purification; D011869:Radioligand Assay; D013972:Thyrotropin / Q000032:analysis
D000596:Amino Acids; D002021:Buffers; D006023:Glycoproteins; D010455:Peptides; D010907:Pituitary Hormones; D011506:Proteins; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone; D013972:Thyrotropin