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

J Immunol Methods. 1990 Jan 24;126(1):135-41. doi: 10.1016/0022-1759(90)90021-m.

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-alpha-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) disfulfide bond between rCD4 and dgA and was able to efficiently kill a human T cell line infected with HIV-1.

Publication types

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

MeSH terms

  • CD4 Antigens / administration & dosage*
  • Chromatography, Liquid
  • Humans
  • Immunotoxins / isolation & purification*
  • Immunotoxins / pharmacology
  • Recombinant Proteins / administration & dosage
  • Ricin / administration & dosage*

Substances

  • CD4 Antigens
  • Immunotoxins
  • Recombinant Proteins
  • Ricin