Preparation of a beta-amanitin-concanavalin A conjugate of low toxicity

Toxicon. 1987;25(9):981-7. doi: 10.1016/0041-0101(87)90161-9.

Abstract

Pure beta-amanitin was isolated by combined adsorption chromatography of a crude methanol extract from Amanita phalloides on Sephasorb HP Ultrafine and Sephadex LH-20. The beta-amanitin was coupled to concanavalin A by the carbodiimide method. The conjugate was purified by fractionation on a column of Sephadex G-75. The molar ratio beta-amanitin to concanavalin A in this conjugate was 4.14. The purified conjugate was tested by thin-layer chromatography and showed characteristic (for amatoxins) bright purple staining but different mobility. The ultraviolet spectrum of the conjugate was different from the spectra of the native beta-amanitin and the native concanavalin A. The toxicity of the conjugate was determined by in vivo toxicological studies and was four-fold lower than that of the native beta-amanitin. These results suggest that this conjugate may be used for immunization procedures and for the production of amatoxin-specific antibodies.

MeSH terms

  • Amanitins* / analysis
  • Amanitins* / toxicity
  • Animals
  • Chromatography, Thin Layer
  • Concanavalin A*
  • Immunodiffusion
  • Spectrophotometry, Ultraviolet

Substances

  • Amanitins
  • Concanavalin A
  • beta-amanitin