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Chip‐based capillary electrophoresis profiling of olive pollen extracts used for allergy diagnosis and immunotherapy
Authors:Agnieszka Zienkiewicz  Krzysztof Zienkiewicz  Fernando Florido  Francisco Manuel Marco  Pedro José Romero  Antonio Jesus Castro  Juan de Dios Alché
Affiliation:1. Department of Biochemistry, Cell and Molecular Biology of Plants, Estación Experimental del Zaidín, CSIC, Granada, Spain;2. Department of Physiology and Molecular Biology of Plants, Nicolaus Copernicus University, Toruń, Poland;3. Department of Cell Biology, Nicolaus Copernicus University, Toruń, Poland;4. Allergy Service, Hospital Universitario “San Cecilio”, Granada, Spain;5. Department of Biotechnology, University of Alicante, Alicante, Spain;6. Department of Medicine, University of Granada, Granada, Spain
Abstract:Standardization of protein extracts for clinical purposes represents an important task in order to maintain adequate reactivity, presence of the relevant allergens, and safety among other factors. The main objective of this work was to explore the potential use of a chip‐based automated CE system commercially available to analyze several of the most common forms of allergenic extracts from olive pollen used in allergy clinics. These include experimental extracts prepared from olive pollens, in‐house reference extracts, extracts designed for skin prick test assays, and a panel of vaccine variants aimed to specific immunotherapy. As a major conclusion of the study, chip‐based CE allowed in all cases to determine accurate protein profiles with different degrees of sensitivity, where several allergens (particularly the major olive pollen allergen Ole e 1) were easily recognized. Moreover, several purified allergens were also analyzed by this method, and proposed as specific standards for different purposes. In the present condition, the method can only provide the protein profile of the extracts with respect to a preestablished standard extract, but not allergen identification. However, these and other future developments and applications are discussed.
Keywords:Allergen  Chip‐Based CE  Extract  Pollen  Standardization  Vaccine
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