Human müllerian inhibiting substance: enhanced purification imparts biochemical stability and restores antiproliferative effects.
Separation of copurifying protease activity from recombinant human Müllerian inhibiting substance (rhMIS) bound to a monoclonal antibody immunoaffinity column by a high-salt wash results in cleaner preparations of rhMIS resistant to cleavage upon storage. In addition, an inhibitor of rhMIS antiproliferative activity is removed. Proteolytic cleavages produced by either a copurifying protease or exogenous plasmin occur at residues 229 and 427 but do not abolish rhMIS biological activity. This report details the modified immunoaffinity column isolation protocol suitable for proteins such as rhMIS and describes the biochemical and antiproliferative properties of this protein.
Ragin
R C
RC
Pediatric Surgical Research Laboratories, Massachusetts General Hospital, Boston.
Donahoe
P K
PK
Kenneally
M K
MK
Ahmad
M F
MF
MacLaughlin
D T
DT
D000595:Amino Acid Sequence; D000818:Animals; D054304:Anti-Mullerian Hormone; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D016466:CHO Cells; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002846:Chromatography, Affinity; D006224:Cricetinae; D010450:Endopeptidases / Q000378:metabolism; D006023:Glycoproteins; D006131:Growth Inhibitors / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007163:Immunosorbent Techniques; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D013696:Temperature; D013735:Testicular Hormones / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D006131:Growth Inhibitors; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D013735:Testicular Hormones; D054304:Anti-Mullerian Hormone; D010450:Endopeptidases