Large scale preparation of immunotoxins constructed with the Fab' fragment of IgG1 murine monoclonal antibodies and chemically deglycosylated ricin A chain

J Immunol Methods. 1988 Sep 13;112(2):267-77. doi: 10.1016/0022-1759(88)90367-5.

Abstract

In this report, we describe a method for the preparation of large amounts (grams) of immunotoxins (ITs) consisting of Fab' fragments of murine IgG1 monoclonal antibodies conjugated to chemically deglycosylated ricin A chain (dgA). The preparation of Fab' and dgA chain and the purification of the Fab'-dgA IT were accomplished by gel filtrations and affinity chromatography utilizing six Pharmacia Bioprocess columns (Sephadex G-25M, Sephacryl S-200HR and Blue Sepharose CL-4B) integrated into a semi-automatic chromatography system controlled by a Pharmacia C3-process controller. The final Fab'-dgA ITs were highly purified, potent, sterile and low in endotoxin concentration.

Publication types

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

MeSH terms

  • Antibodies, Monoclonal*
  • Antigens, CD*
  • Antigens, Differentiation, B-Lymphocyte / immunology*
  • Biotechnology / methods
  • Cell Adhesion Molecules*
  • Chemical Phenomena
  • Chemistry, Physical
  • Chromatography
  • Endotoxins / analysis
  • Glycoproteins
  • Hydrogen-Ion Concentration
  • Immunoglobulin Fab Fragments
  • Immunoglobulin G
  • Immunotoxins*
  • Lectins*
  • Oxidation-Reduction
  • Ricin*
  • Sialic Acid Binding Ig-like Lectin 2

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • Antigens, Differentiation, B-Lymphocyte
  • Cell Adhesion Molecules
  • Endotoxins
  • Glycoproteins
  • Immunoglobulin Fab Fragments
  • Immunoglobulin G
  • Immunotoxins
  • Lectins
  • Sialic Acid Binding Ig-like Lectin 2
  • Ricin