A Promising Technology for Characterizing Proteins, Antigens and Allergens in Extracts and Source Materials
Saturday, March 5, 2016
South Exhibit Hall H (Convention Center)
Jack D. Kelly
Rationale: Genetic and environmental factors can affect the compositions of allergenic extracts and can lead to variabilities in proteins, antigens, and allergens from one product lot to another. Traditional characterization methods (SDS-PAGE/Western blotting) have not been conducive for high throughput screening of allergenic extracts and source materials. A new technology can facilitate the production of these profiles. The objectives herein are: 1) demonstrate the application of this technology (“WES”) for source material assessments, 2) demonstrate that both WES and traditional techniques provide qualitatively similar protein and antigenic profiles, and 3) illustrate the quantitative capabilities of WES.

Methods: Source materials and allergenic extracts were evaluated for total protein (Bradford) and protein/antigenic profiling (WES Nanotechnology Capillary Electrophoresis from ProteinSimple, Santa Clara, CA, or conventional SDS-PAGE / Western Blotting).  WES kits contain sample preparation reagents, pre-filled plates with proprietary electrophoretic reagents, disposable capillary cartridges, and all post-sample preparation steps (loading, washing, immunoprobing, data analysis, incubation time) are fully automated.

Results: Both protein and antigenic patterns were qualitatively similar between WES and the traditional techniques. WES provided several additional benefits, including small reagent volume (5-10 µL), quantitation, high throughput (3-4 hour run times), complete automation of reagents and assay conditions, analytical integrations, matrix-based molecular weight determination, and real-time migration visualizations.

Conclusions: WES assays were simple to perform and provided both similar qualitative data compared to traditional methods and useful quantitative data with full automation. This new nanotechnology allows for a higher throughput of raw material and allergenic extract samples using 5-10 µL of each reagent.