Preparation and characterization of the F (ab)2 fragments of an aromatase activity-suppressing monoclonal antibody

Steroids. 1997 Dec;62(12):776-81. doi: 10.1016/s0039-128x(97)00090-1.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antibodies, Monoclonal / isolation & purification
  • Antibodies, Monoclonal / pharmacology
  • Aromatase / drug effects
  • Aromatase / immunology
  • Aromatase / metabolism*
  • Humans
  • Immunoglobulin Fab Fragments / isolation & purification
  • Immunoglobulin Fab Fragments / metabolism*
  • Immunoglobulin Fab Fragments / pharmacology*
  • Immunoglobulin Fragments / isolation & purification
  • Immunoglobulin Fragments / pharmacology
  • Papain / chemistry
  • Papain / metabolism

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin Fab Fragments
  • Immunoglobulin Fragments
  • Aromatase
  • Papain