不同方法提取和处理中华绒螯蟹组织蛋白的过敏原组分分析

发布时间：2018-06-25 13:38

本文选题：中华绒螯蟹 + 食物过敏　；参考：《中国免疫学杂志》2014年12期

【摘要】：目的:分析不同方法提取和处理中华绒螯蟹组织的蛋白组分及过敏原组分,为检测中华绒螯蟹特异性IgE提供最佳抗原制备方法。方法:分别采用PBS提取法、丙酮抽提法和裂解液提取法提取中华绒螯蟹组织蛋白,并用裂解液提取法分别提取加热前、后的蟹蛋白;采用SDS-PAGE和双向电泳分析蛋白组分,采用Western blot分析过敏原组分。结果:不同方法提取的蛋白组分、sIgE与蛋白结合的强度和所识别的蛋白组分存在一定差异。PBS提取液蛋白主要分布在94、85、76、66、18kD,丙酮提取液蛋白主要分布在94、85、76、36 kD,裂解液提取液蛋白主要分布在200、125、51、43、38 kD。PBS提取液与sIgE结合的阳性反应条带主要分布在76、66、53、43、38、18 kD;丙酮提取液主要分布在94、76、66、50、43、38、18 kD;裂解液提取液主要分布在200、105、94、76、43、38、18 kD。加热前、后蟹蛋白过敏原组分差别不大。结论:裂解液提取法提取的中华绒螯蟹过敏原更适于制备测定特异性IgE的抗原。加热不影响蟹过敏原组分和与特异性IgE的结合活性。
[Abstract]:Aim: to analyze the protein and allergen components of Chinese mitten crab (Eriocheir sinensis) extracted and treated by different methods, and to provide the best preparation method for detecting specific IgE of Eriocheir sinensis (Eriocheir sinensis). Methods: the tissue proteins of Eriocheir sinensis were extracted by PBS extraction, acetone extraction and lytic solution extraction respectively. The allergen components were analyzed by Western blot. Results: there were some differences in the binding strength between Siga E and protein and the protein components recognized by different methods. The protein of PBS extract was mainly distributed in 94 / 85 / 766 / 66 / 666 / 18kD. the protein of acetone extract was mainly distributed in 94 / 85 / 767 / 636kD, and the cleavage solution was mainly distributed in the solution of cleavage. The protein was mainly distributed in the SIgE binding positive reaction bands of the extracted solution of PBS from 2000.125A, 51kD.PBS and Siga, and the acetone solution was mainly distributed in 94A125C7506333818kD, and the cleavage solution was mainly distributed in 2000105C9105C944333388kD, and the protein was mainly distributed in the 94A125C 650333818kDand the acetone extract was mainly distributed in 94105Dand 73333388kD.The protein was mainly distributed in the extract of PBS, which was bound to Siga E, and the protein was mainly distributed in 766. Before heating, there was no significant difference in the components of protein allergens in crab. Conclusion: the allergen of Eriocheir sinensis extracted by lytic solution is more suitable for the preparation of specific IgE antigen. Heating did not affect the allergen composition and specific IgE binding activity of crab.
【作者单位】：天津医科大学医学检验学院;天津市中医药研究院附属医院;
【基金】：国家自然基金项目资助项目(81371882)
【分类号】：R392.8

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