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目的建立一种简捷、准确、快速检测食品过敏原的方法。分析河虾总蛋白2-DE(2-DimensionElectrophoresis)指纹图谱的重现性,确定食品过敏原指纹图谱快速检测的可行性。方法新鲜河虾,去头和虾皮,搅碎后丙酮去脂,PBS抽提总蛋白,冻干后为初提样品,试剂盒精提,双向电泳IEF选择11cm和13cm两种不同厂家、规格的非线性胶条(pH=3～10)溶胀上样,平衡后转移胶条在不同厂家、型号的电泳槽中进行第二向SDS-PAGE垂直电泳(10%),银染和考染后扫描图像进行计算机分析处理。结果2-DE谱图显示河虾总蛋白分布较广,其高丰度蛋白主要是一些大分子量酸性蛋白。不同电泳系统,选用不同规格的IPG胶条所得河虾总蛋白2-DE指纹谱图具有良好的重现性。银染灵敏度较高,更适合食品过敏原指纹图谱的快速检测。结论食品过敏原总蛋白2-DE指纹图谱具有良好的重现性和稳定性,可以用于食品过敏原的快速检测。
Objective To establish a simple, accurate and rapid detection of food allergens. The reproducibility of 2-DE (2-Dimension Electrophoresis) fingerprinting was analyzed to determine the feasibility of rapid detection of food allergen fingerprinting. Methods Fresh shrimp, go head and shrimp skin, crushed acetone degreasing, PBS extraction of total protein, after lyophilization as a preliminary sample, kit refinement, two-dimensional electrophoresis IEF choose 11cm and 13cm two different manufacturers, specifications Non-linear strips (pH = 3 ~ 10) were swollen for loading. Balanced strips were transferred to second-dimension SDS-PAGE (10%) in different manufacturers and models. Silver stain and post- Image processing computer analysis. Results The 2-DE spectrum showed that the total protein of shrimp was widely distributed, and the high-abundance proteins were mainly some large-molecular-weight acidic proteins. Different electrophoresis systems, using different specifications of IPG strips of shrimp total protein 2-DE fingerprints with good reproducibility. High silver staining sensitivity, more suitable for rapid detection of food allergen fingerprinting. Conclusion The total protein 2-DE fingerprint of food allergen has good reproducibility and stability and can be used for the rapid detection of food allergens.