Sunday, 6 December 2015
Ivanka Trump Ballroom (Trump National Doral)
Introduction: Allergen extracts are needed for the specific diagnosis of allergic sensitization to foods. Egg white, wheat, soy, peanuts and shrimp are common food in Cuba causing allergic symptoms. Glycerol is commonly used in allergen extract formulation as stabilizing agent for protein components. The aim of this work was to assess the precision, accuracy, detection limit and specificity of the SDS-Polyacrilamide Gel Electrophoresis technique in determining the protein profile if glycerol formulated allergen extracts of these foods. Materials and Methods: SDS-PAGE technique was applied to allergen extracts of egg white, soy, wheat, peanut and shrimp, manufactured in BIOCEN, Cuba, formulated in 50% glycerol. Major allergens bands were identified by comparing the electrophoresis patterns with the IgE binding bands as measured by western blotting. Light scanning densitometry was used for semiquantitative assessment and molecular weight determination. Possible glycerol interference was determined compared to the lyophilized references. Results and Discussion: All pre-established acceptance criteria he analytical technique were met. Coefficient of variation (CV) was less than 5% in all cases. There were no significant differences between different analysts, or between intra-assay and inter-assay variability (Fischer test, p<0.05). Accuracy in the determination of molecular weights was demonstrated showing no significant differences to the theoretical values for major allergens (p=0.05). The lack of interference of glycerol in the final formulation was also demonstrated, since no significant differences in the percentages of the major bands were noted between reconstituted lyophilized allergen extract and products containing glycerol (p=0.05). The detection limit of the technique was 1µg for Wheat, Peanut and Soy, 2 µg for shrimp and 4µg for Egg White. Conclusions: The SDS PAGE technique showed to be appropriate, with suitable precision, accuracy and detection limit for assessing the protein composition of glycerinated allergen extracts of several food allergenic sources.
Learning Objectives:
The aim of this work was to assess the precision, accuracy, detection limit and specificity of the SDS-Polyacrilamide Gel Electrophoresis technique in determining the protein profile if glycerol formulated allergen extracts of Egg white, wheat, soy, peanuts and shrimp, used for the specific diagnosis of allergic sensitization.
The aim of this work was to assess the precision, accuracy, detection limit and specificity of the SDS-Polyacrilamide Gel Electrophoresis technique in determining the protein profile if glycerol formulated allergen extracts of Egg white, wheat, soy, peanuts and shrimp, used for the specific diagnosis of allergic sensitization.