Thiophilic adsorption chromatography: purification of Equ c2 and Equ c3, two horse allergens from horse sweat

J Chromatogr B Biomed Sci Appl. 1998 Jun 12;710(1-2):57-65. doi: 10.1016/s0378-4347(98)00130-3.

Abstract

Purification of two allergens from horse (Equus caballus) sweat, Equ c2 and Equ c3, by means of salt-promoted chromatography on a "thiophilic" (T-gel) adsorbent is described. Immobilization of these proteins was found to be dependent on the presence of water-structure-forming salts where the ammonium sulphate concentration in the equilibration buffer was 2 M. Equ c2 showed higher affinity towards the thiophilic matrix than Equ c3. Their molecular mass (Mr) values established by SDS-polyacrylamide gel electrophoresis were for Equ c2 approximately 17,000 and for Equ c3 approximately 16,000, and both proteins showed a low isoelectric point of approximately 3.8. Their allergenic properties were also investigated using sera from horse-sensitized patients, where it was demonstrated that these proteins exhibited an IgE antibody binding capacity. In this report we show the broad potential applications of thiophilic adsorption chromatography for the efficient purification of allergens.

MeSH terms

  • Adsorption
  • Allergens / immunology
  • Allergens / isolation & purification*
  • Animals
  • Chromatography / methods*
  • Electrophoresis, Agar Gel
  • Horses* / immunology
  • Humans
  • Hypersensitivity / immunology
  • Sulfhydryl Reagents
  • Sweat / chemistry*
  • Sweat / immunology

Substances

  • Allergens
  • Sulfhydryl Reagents