Enhanced solubilization of immunoreactive proteins from Brugia malayi adult parasites using cetyltrimethylammonium bromide.
To determine an optimal method for extracting immunoreactive proteins from filarial parasites, we have subjected Brugia malayi adult worms to a variety of solubilization regimens and compared the results. The parasites were extracted in one of seven detergents (including anionic, cationic, nonionic, and zwitterionic compounds) under varying conditions of pH, detergent concentrations, and incubation time. The individual antigen preparations were then compared both by one-dimensional SDS-PAGE and by immunoblotting analysis using a serum pool from individuals resident in an area endemic for lymphatic filariasis. The cationic detergent cetyltrimethylammonium bromide (CTAB) at 1.0% concentration, pH 7.2, consistently solubilized more proteins immunoreactive with the sera tested. Additionally, CTAB never failed to solubilize immunoreactive proteins solubilized by those other detergents or combinations of detergents studied.
Freedman
D O
DO
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.
Nutman
T B
TB
Ottesen
E A
EA
D000818:Animals; D000947:Antigens, Helminth / Q000032:analysis / Q000302:isolation & purification*; D002009:Brugia / Q000276:immunology*; D000077286:Cetrimonium; D002593:Cetrimonium Compounds; D002793:Cholic Acids; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D000644:Quaternary Ammonium Compounds; D012995:Solubility; D013501:Surface-Active Agents
D000947:Antigens, Helminth; D002593:Cetrimonium Compounds; D002793:Cholic Acids; D003902:Detergents; D000644:Quaternary Ammonium Compounds; D013501:Surface-Active Agents; C048531:chapso; D000077286:Cetrimonium