坚果类过敏原的分离及免疫印迹分析

Isolation of Allergens in Nuts and Analysis on Western Blotting

摘要:

本文采用传统的丙酮、乙醚溶剂去脂方法，选用不同品种和不同种类的坚果食品对其进行过敏蛋白的初步分离，应用BCA蛋白试剂盒测定各蛋白浓度，通过SDS-PAGE电泳分离坚果过敏蛋白各组份，采用免疫印迹(Western-blotting)方法鉴定坚果蛋白的过敏性。结果表明，各坚果蛋白提取浓度为白芝麻：14 mg/mL，黑芝麻：4 mg/mL，核桃：8 mg/mL，腰果：28 mg/mL，开心果：21 mg/mLL。SDS-PAGE电泳显示15 ku是白芝麻的主要蛋白，34 ku是黑芝麻的主要蛋白，19和37 ku是腰果的主要蛋白，13 ku是核桃的主要蛋白，13和21 ku是开心果的主要蛋白。Western-Blotting 显示对过敏患者的阳性混合血清均有过敏反应，坚果过敏具有同源性，白芝麻与黑芝麻的主要过敏原分子量均在22 ku，腰果的主要过敏原分子量在20 ku，核桃的主要过敏原分子量在54 ku，开心果的主要过敏原分子量在23 ku。

Abstract:

Acetone and ethyl ether solvent were used to degrease of different kinds of nuts, then the allergen were isolated from the defatted nuts. The concentration and molecular weight of allergenic proteins were tested by the BCA protein kit and SDS-PAGE. Western –blotting was adopted for the identification of nut allergens. Results showed that the extracted concentration of the protein of white sesame, black sesame seed, walnut, cashew and pistachios were 14 mg/mL, 4 mg/mL, 8 mg/mL, 28 mg/ mL and 21 mg/ mL, respectively. In addition, the molecular weight of each allergenic protein from the obove-mentioned five nuts were 15 ku, 34 ku, 19 and 37 ku, 13 ku, 13 and 21 ku, respectively. The Western - blotting method showed allergenic reaction to patients with positive mixed serum. Nut allergy had homology, and the main allergens of sesame, cashew nuts, walnut and pistachios were at 22, 20, 54 and 23 ku.