a protein allergen microarray detects specific ige to pollen surface, cytoplasmic, and commercial allergen extracts过敏原蛋白质微阵列检测特定的ige花粉表面,细胞质,商业过敏原提取物.pdfVIP

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2017-09-05 发布于上海
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a protein allergen microarray detects specific ige to pollen surface, cytoplasmic, and commercial allergen extracts过敏原蛋白质微阵列检测特定的ige花粉表面,细胞质,商业过敏原提取物.pdf

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a protein allergen microarray detects specific ige to pollen surface, cytoplasmic, and commercial allergen extracts过敏原蛋白质微阵列检测特定的ige花粉表面,细胞质,商业过敏原提取物

A Protein Allergen Microarray Detects Specific IgE to Pollen Surface, Cytoplasmic, and Commercial Allergen Extracts 1 ¤a 1¤b 2¤c Katinka A. Vigh-Conrad * , Donald F. Conrad , Daphne Preuss 1 Department of Human Genetics, The University of Chicago, Chicago, Illinois, United States of America, 2 Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, Illinois, United States of America Abstract Background: Current diagnostics for allergies, such as skin prick and radioallergosorbent tests, do not allow for inexpensive, high-throughput screening of patients. Additionally, extracts used in these methods are made from washed pollen that lacks pollen surface materials that may contain allergens. Methodology/Principal Findings: We sought to develop a high-throughput assay to rapidly measure allergen-specific IgE in sera and to explore the relative allergenicity of different pollen fractions (i.e. surface, cytoplasmic, commercial extracts). To do this, we generated a protein microarray containing surface, cytoplasmic, and commercial extracts from 22 pollen species, commercial extracts from nine non-pollen allergens, and five recombinant allergenic proteins. Pollen surface and cytoplasmic fractions were prepared by extraction into organic solvents and aqueous buffers, respectively. Arrays were incubated with ,25 uL of serum from 176 individuals and bound IgE was detected by indirect immunofluorescence, providing a high- throughput measurement of IgE. We demonstrated that the allergen microarray is a reproducible method to measure allergen-specific IgE in small amounts of sera. Using this tool, we demonstrated that specific IgE clusters according to the phylogeny of the