花生、河虾中主要过敏原组分的研究

Main allergen components of peanut and fresh water shrimp

目的鉴定并纯化花生、河虾中引起过敏原反应的主要蛋白组分。方法花生、河虾蛋白浸液腹腔注射建立小鼠过敏模型，用硫酸铵分级沉淀、等电点沉淀纯化目的蛋白，建立问接ELISA检测抗血清中的特异性IgE水平。结果SDS-PAGE鉴定纯化后的花生蛋白中相对分子量63．5kDa、60kDa、37kDa、17．5kDa，虾蛋白中相对分子量36kDa、85kDa、52kDa为主要过敏原组分。将纯化后的蛋白注射BALB／C小鼠腹腔，获得含特异性IgE的小鼠血清进行研究。棋盘试验表明花生蛋白抗原最佳稀释度为1：300-1：600，抗血清稀释度为1：3000-1：5000，河虾蛋白抗原最佳稀释度为1：200-1：500，抗血清稀释度为1：600-1：1000。结论成功获得花生、河虾中主要过敏原组分及其血清抗体，为过敏原的分析研究提供了一套比较完整的方法。

Objective To identify and purify main protein components in peanut and fresh water shrimp that can cause allergic reactions. Methods Experimental allergic model was produced in mice by intraperitoneal injection of extract of peanut and fresh water shrimp. The target protein was isolated and purified by ammonium sulfate fractionation and isoelectric point precipitation. Serum specific immunoglobulin E （IgE） for food allergen in model mice was measured by indirect enzyme-linked immunosorbent assay （ELISA）, Results Relative molecular weight of main peanut protein after purification was 63.5 kDa, 60 kDa, 37 kDa and 17.5 kDa, respectively, by SDS-PAGE, and that of fresh water shrimp was 36 kDa, 85 kDa and 52 kDa, respectively. BALB/C mice were injected with purified protein allergen and high titer of serum IgE was isolated from them for detection. Chessboard experiments suggested that the optimal dilution of peanut protein allergen was 1 ： 300 - 1 ： 600, with antiserum dilution of 1 ： 3 000 - 1 ： 5 000, and that of shrimp protein was 1 ： 20 - 1 ： 500, with antiserum dilution of 1 ： 600 - 1 ： 1 000. Conclusion Main allergen components of peanut and fresh water shrimp and their antibodies were successfully obtained in this study, which provide a more comprehensive approach for allergen analysis.