Purification of human pituitary LH and thyrotrophin by hydrophobic chromatography.
A new preparative procedure is described for the efficient purification of LH and TSH from frozen human pituitary glands. LH and TSH were isolated simultaneously from crude preparations by hydrophobic-interaction chromatography followed by separation and purification by reverse-phase high-performance liquid chromatography in a single step. Highly purified hormones were prepared in good yields (45 mg LH and 20 mg TSH/1000 glands) and with high biological activities; the potencies of purified LH and TSH were 5.8 x NIH-LH-S1 equivalent in an ovarian ascorbic acid depletion assay and 7.1 U/mg against human TSH MRC Research Standard (T1 70/9) in the McKenzie assay respectively. Cross-contamination by other glycoprotein hormones was low.
Hiyama
J
J
Department of Biochemistry, University of Auckland, New Zealand.
Surus
A
A
Renwick
A G
AG
D000818:Animals; D001681:Biological Assay / Q000379:methods; D002845:Chromatography / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D007986:Luteinizing Hormone / Q000302:isolation & purification*; D051379:Mice; D010902:Pituitary Gland / Q000032:analysis*; D011863:Radioimmunoassay / Q000379:methods; D013972:Thyrotropin / Q000302:isolation & purification*
D007986:Luteinizing Hormone; D013972:Thyrotropin