Large scale preparation of an immunoconjugate constructed with human recombinant CD4 and deglycosylated ricin A chain

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

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

A method for the preparation and purification of large amounts (grams) of a conjugate containing recombinant CD4 antigen (rCD4) and chemically deglycosylated ricin A chain (dgA) is described. The cross-linking of rCD4 and dgA molecules was accomplished with N-succinimidyl-oxycarbonyl-α-methyl-(2-pyridyldithio)toluene (SMPT). The rCD4-dgA conjugate was purified by an automatic liquid chromatography system consisting of Blue Sepharose CL-4B and Sephacryl S-200HR Pharmacia Bioprocess columns. The purified, endotoxin-free rCD4-dgA conjugate had a stable (hindered) disulfide bond between rCD4 and dgA and was able to efficiently kill a human T cell line infected HIV-1.

Original languageEnglish (US)
Pages (from-to)135-141
Number of pages7
JournalJournal of Immunological Methods
Volume126
Issue number1
DOIs
StatePublished - Jan 24 1990

Keywords

  • CD4
  • Human immunodeficiency virus
  • Immunoconjugate
  • Ricin A chain

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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