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

Victor Ghetie, Philip Thorpe, Maria Ana Ghetie, Phillip Knowles, Jonathan W. Uhr, Ellen S. Vitetta

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)223-230
Number of pages8
JournalJournal of Immunological Methods
Volume142
Issue number2
DOIs
StatePublished - Sep 13 1991

Keywords

  • Deglycosylated ricin A chain
  • Good laboratory practice
  • Immunotoxin
  • Large scale
  • Monoclonal antibody

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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