School of Chemical Engineering, Food Science and Technology, University of New South Wales, Sydney, NSW 2052, Australia.
J Agric Food Chem. 2012 Oct 17;60(41):10352-8. doi: 10.1021/jf302800e. Epub 2012 Oct 9.
This study aimed to purify and characterize the peanut allergens Ara h1 and Ara h3 from four cultivars that represent the four major market types to provide better understanding of the molecular organization of oligomers in different market types. The chromatographic profiles of Ara h1 and Ara h3 from the four cultivars obtained from anion exchange chromatography were similar. However, they differed in the distribution of trimeric and hexameric structures of Ara h3 isolated by size exclusion chromatography. The Menzies (Runner market type) and Walter (Spanish market type) cultivars, wherein Ara h3 proteins consist of two acidic subunits, exhibited trimeric and hexameric conformations proportionally. However, the Middleton (Virginia market type) and Kelinci (Valencia market type) cultivars, wherein Ara h3 proteins consist of three acidic subunits, showed predominantly a hexameric structure. The oligomeric structures of the purified Ara h1 demonstrated strong IgE binding properties, whereas the allergenic property of the oligomeric Ara h3 could not be performed due to lack of availability of specific IgE. In addition, the polyclonal antibodies raised against the purified Ara h1 and Ara h3 showed highly specific binding to their respective antigens.
本研究旨在从四个代表四种主要市场类型的品种中纯化和鉴定花生过敏原 Ara h1 和 Ara h3，以更好地了解不同市场类型中寡聚物的分子结构。从阴离子交换层析获得的四个品种的 Ara h1 和 Ara h3 的色谱图谱相似，但通过大小排阻层析分离的 Ara h3 的三聚体和六聚体结构的分布不同。Menzies（Runner 市场类型）和 Walter（Spanish 市场类型）品种中，Ara h3 蛋白由两个酸性亚基组成，呈现出成比例的三聚体和六聚体构象。然而，Middleton（Virginia 市场类型）和 Kelinci（Valencia 市场类型）品种中，Ara h3 蛋白由三个酸性亚基组成，主要呈现六聚体结构。纯化的 Ara h1 的寡聚体结构表现出强烈的 IgE 结合特性，而由于缺乏特异性 IgE，无法对寡聚 Ara h3 的变应原性进行检测。此外，针对纯化的 Ara h1 和 Ara h3 产生的多克隆抗体对各自的抗原表现出高度特异性结合。
