The GLP large scale preparation of immunotoxins containing deglycosylated ricin A chain and a hindered disulfide bond

J Immunol Methods. 1991 Sep 13;142(2):223-30. doi: 10.1016/0022-1759(91)90110-2.

Abstract

The large scale preparation of two second generation immunotoxins containing murine monoclonal antibodies and deglycosylated ricin A chain is described. The procedure for the preparation of immunotoxins consists of the derivatization of antibody with SMPT and reduction of dgA with DTT followed by their reaction to establish a hindered interchain disulfide bond. The purification of the immunotoxin includes affinity chromatography on Blue-Sepharose to remove the free antibody and gel filtration on Sephacryl S-200HR to remove any high molecular weight material and free dgA. The two immunotoxins were prepared by GLP procedures and tested for yield, composition, purity, sterility and biological activity.

Publication types

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

MeSH terms

  • Antibodies, Monoclonal
  • Antigens, CD / immunology
  • Antigens, CD19
  • Antigens, Differentiation, B-Lymphocyte / immunology
  • Cell Adhesion Molecules*
  • Chromatography, Gel
  • Chromatography, High Pressure Liquid
  • Cross-Linking Reagents
  • Cysteine
  • Electrophoresis, Polyacrylamide Gel
  • Immunotoxins*
  • Lectins*
  • Ricin
  • Sialic Acid Binding Ig-like Lectin 2
  • Succinimides

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • Antigens, CD19
  • Antigens, Differentiation, B-Lymphocyte
  • Cell Adhesion Molecules
  • Cross-Linking Reagents
  • Immunotoxins
  • Lectins
  • Sialic Acid Binding Ig-like Lectin 2
  • Succinimides
  • 4-succinimidyloxycarbonyl-alpha-methyl-alpha(2-pyridyldithio)toluene
  • Ricin
  • Cysteine