Nanobiotechology and Bioanalysis Group, Departament d'Enginyeria Quimica, Universitat Rovira i Virgili, Avinguda Països Catalans 26, 43007 Tarragona, Spain.
J Agric Food Chem. 2011 Mar 23;59(6):2752-8. doi: 10.1021/jf104062d. Epub 2011 Feb 18.
Lupin has recently been added to the list of allergens requiring mandatory advisory labeling on foodstuffs sold in the European Union, and since December 2008, all products containing even trace amounts of lupin must be labeled correctly. Lupin globulins consist of two major globulins called α-conglutin (11S and "legumin-like") and β-conglutin (7S and "vicilin-like") and another additional two globulins, γ-conglutin and δ-conglutin, which are present in lower amounts. We report on a methodology to facilitate the extraction of each of these proteins using centrifugation and isolation by anion-exchange chromatography followed by size-exclusion chromatography. The isolated subunits were characterized using reducing and non-reducing polyacrylamide gel electrophoresis, western blotting, and peptide mass fingerprinting, all of which revealed that the individual protein subunits are highly pure and can be used as immunogens for the production of antibodies specific for each of the conglutin fractions, as well as standards, and the extraction protocol can be used for the selective extraction of each of the subunits from foodstuffs, thus facilitating a highly accurate determination of the lupin concentration. Furthermore, the subunits can be used to elucidate information regarding the toxicity of each of the subunits, by looking at their interaction with the IgE antibodies found in the serum of individuals allergic to lupin, providing critical information for the definition of the requirements of analytical assays for the detection of lupin in foodstuffs.
羽扇豆最近被添加到需要在欧盟销售的食品中进行强制性咨询标签的过敏原列表中，自 2008 年 12 月以来，所有含有羽扇豆痕量的产品都必须正确贴上标签。羽扇豆球蛋白由两种主要球蛋白组成，称为α-伴球蛋白（11S 和“豆球蛋白样”）和β-伴球蛋白（7S 和“vicilin 样”），以及另外两种球蛋白，γ-伴球蛋白和δ-伴球蛋白，它们的含量较低。我们报告了一种使用离心和阴离子交换色谱分离来提取这些蛋白质的方法，然后进行大小排阻色谱。使用还原和非还原聚丙烯酰胺凝胶电泳、western 印迹和肽质量指纹图谱对分离的亚基进行了表征，所有这些都表明各个蛋白质亚基高度纯，可以用作针对各伴球蛋白部分产生抗体的免疫原，也可以用作标准品，并且提取方案可用于从食品中选择性提取各个亚基，从而可以非常准确地确定羽扇豆的浓度。此外，通过观察它们与对羽扇豆过敏的个体血清中的 IgE 抗体的相互作用，可以阐明每个亚基的毒性信息，从而为确定食品中羽扇豆的分析检测要求提供关键信息。
