Isolation of the 36-kD German (Blattella germanica) cockroach allergen using fast protein liquid chromatography

Int Arch Allergy Immunol. 1994;103(1):59-66. doi: 10.1159/000236606.

Abstract

Cockroach allergens have been established as important sensitizing agents in the induction of asthma. In the present investigation, we identified a 36-kD allergen from German cockroach (Blattella germanica) using fast protein liquid chromatography that reacted in the Bla g II monoclonal assay developed by Pollart et al. Defatted whole body German cockroaches were extracted in phosphate-buffered saline and fractionated using gel filtration (Superdex 75) and anion exchange (MonoQ) chromatography. The 36-kD allergen was isolated from this enriched allergen preparation by excision from modified preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels, electroelution, dialysis and lyophilization. SDS-PAGE/Western immunoblot analysis using a serum pool of cockroach-sensitive individuals revealed a single band. The lyophilized 36-kD allergen was then analyzed for amino acid composition and digested for peptide amino acid sequencing.

Publication types

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

MeSH terms

  • Allergens / isolation & purification*
  • Amino Acid Sequence
  • Amino Acids / analysis
  • Animals
  • Antibody Specificity
  • Chromatography, Gel
  • Chromatography, Liquid / methods
  • Cockroaches / immunology*
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Immunoglobulin E / immunology
  • Molecular Sequence Data
  • Sodium Dodecyl Sulfate

Substances

  • Allergens
  • Amino Acids
  • Sodium Dodecyl Sulfate
  • Immunoglobulin E