Preparation and characterization of the F (ab)2 fragments of an aromatase activity-suppressing monoclonal antibody.
The preparation and characterization of the Fab and F(ab')2 fragments of a murine monoclonal antibody specific for aromatase cytochrome P-450 and which is suppressive of estrogen biosynthesis are described. This monoclonal antibody, MAb3-2C2, was purified from murine ascites using protein A affinity chromatography and digested with immobilized papain to produce antibody fragments. The Fab and F(ab')2 fragments were then purified using protein A affinity chromatography and S-200 HR size exclusion chromatography. The Fab fragment was further purified using S-100 HR size exclusion chromatography. Both the Fab and F(ab')2 fragments of the MAb3-2C2 suppressed aromatase activity in a dose-dependent manner. While the F(ab')2 fragment (110 kDa) maintained potent suppressive activity, the Fab fragment (42 kDa) required a higher concentration to suppress aromatase activity as compared to the IgG.
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Endocrine Biochemistry Department, Hauptman-Woodward Medical Research Institute, Buffalo, NY 14203-1196, USA.
Osawa
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D000911:Antibodies, Monoclonal / Q000302:isolation & purification / Q000494:pharmacology; D001141:Aromatase / Q000187:drug effects / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology*; D007128:Immunoglobulin Fragments / Q000302:isolation & purification / Q000494:pharmacology; D010206:Papain / Q000737:chemistry / Q000378:metabolism
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007128:Immunoglobulin Fragments; D001141:Aromatase; D010206:Papain