New approaches for high-yield purification of Müllerian inhibiting substance improve its bioactivity

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 766; no. 1; pp. 89 - 98
• Main Authors: Lorenzo, Hans K, Teixeira, Jose, Pahlavan, Nima, Laurich, V.Matt, Donahoe, Patricia K, MacLaughlin, David T
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 05.01.2002
• Subjects: Animals; Anti-Mullerian Hormone; CHO Cells; Chromatography, Affinity - methods; Chromatography, Ion Exchange - methods; Cricetinae; Glycoproteins; Growth Inhibitors - isolation & purification; Growth Inhibitors - pharmacology; Humans; Müllerian inhibiting substance; Recombinant Proteins - isolation & purification; Recombinant Proteins - pharmacology; Testicular Hormones - isolation & purification; Testicular Hormones - pharmacology; Transforming growth factor β; Protein purification; Transforming growth factor β; Müllerian inhibiting substance
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/S0378-4347(01)00436-4

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 β family with which MIS shares sequence homology.