Isolation of the 36-kD German (Blattella germanica) cockroach allergen using fast protein liquid chromatography.
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.
Helm
R M
RM
Department of Pediatrics, University of Arkansas for Medical Sciences, Arkansas Children's Hospital, Little Rock 72202.
Brenner
R J
RJ
Williams
L W
LW
Burks
A W
AW
D000485:Allergens / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000918:Antibody Specificity; D002850:Chromatography, Gel; D002853:Chromatography, Liquid / Q000379:methods; D003058:Cockroaches / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D007073:Immunoglobulin E / Q000276:immunology; D008969:Molecular Sequence Data; D012967:Sodium Dodecyl Sulfate
D000485:Allergens; D000596:Amino Acids; D012967:Sodium Dodecyl Sulfate; D007073:Immunoglobulin E