One-step procedure for the rapid isolation of mouse monoclonal antibodies and their antigen binding fragments by fast protein liquid chromatography on a mono Q anion-exchange column

J Chromatogr. 1985 Jan 25;319(1):67-77. doi: 10.1016/s0021-9673(01)90540-0.

Abstract

A one-step chromatographic procedure was used to isolate rapidly mouse IgG monoclonal antibodies (mAbs) (IgG1, IgG2a, IgG2b) contained in ascites fluids and Fab fragments contained in papain-treated mAb suspensions. Chromatographic separations were performed on an anion-exchange Mono Q column connected to a fast protein liquid chromatographic (FPLC) system. Detection of mAb or their antigen binding fragments (Fab) in eluted peaks was performed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis together with a silver or a Coomassie Brillant Blue R 250 staining technique and solid phase radioimmunoassay with 125I-labelled sheep anti-mouse antibodies directed against total immunoglobulins. Rapid assessment of the purity of isolated mAbs and their Fab fragments was performed by gel permeation chromatography on a TSK G 3000 SW column. Mouse mAbs and their Fab fragments were rapidly isolated (25 min), in a functionally active state, to a high degree of purity on the FPLC-Mono Q system compared to the time taken by other techniques.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / isolation & purification*
  • Binding Sites, Antibody
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Chromatography, Liquid / methods
  • Electrophoresis, Polyacrylamide Gel
  • Mice
  • Radioimmunoassay
  • Sodium Dodecyl Sulfate

Substances

  • Antibodies, Monoclonal
  • Sodium Dodecyl Sulfate