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

V. Ghetie, M. A. Ghetie, J. W. Uhr, E. S. Vitetta

Research output: Contribution to journalArticle

25 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)267-277
Number of pages11
JournalJournal of Immunological Methods
Volume112
Issue number2
DOIs
StatePublished - Sep 13 1988

Keywords

  • Fab′ fragment
  • Immunotoxin
  • Large scale preparation of proteins
  • Monoclonal antibody
  • Ricin toxin deglycosylated A chain

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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