New approaches for high-yield purification of Müllerian inhibiting substance improve its bioactivity.
We have established a new method to purify Müllerian inhibiting substance (MIS) with higher purity and recovery over existing procedures. Recombinant human MIS was expressed in Chinese hamster ovary cells and secreted into chemically defined serum-free media. The secreted products were concentrated by either precipitation with ammonium sulfate or lectin-affinity chromatography, each of which was followed by anion-exchange chromatography. Further separation of the active carboxy-terminal domain of MIS was achieved after cleavage by plasmin followed by lectin-affinity chromatography. This method may be applicable to other members of the transforming growth factor beta family with which MIS shares sequence homology.
Lorenzo
Hans K
HK
Massachusetts General Hospital and the Department of Surgery, Harvard Medical School, Boston 02114, USA.
Teixeira
Jose
J
Pahlavan
Nima
N
Laurich
V Matt
VM
Donahoe
Patricia K
PK
MacLaughlin
David T
DT
D000818:Animals; D054304:Anti-Mullerian Hormone; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D006224:Cricetinae; D006023:Glycoproteins; D006131:Growth Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D006801:Humans; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000494:pharmacology; D013735:Testicular Hormones / Q000302:isolation & purification* / Q000494:pharmacology
D006023:Glycoproteins; D006131:Growth Inhibitors; D011994:Recombinant Proteins; D013735:Testicular Hormones; D054304:Anti-Mullerian Hormone