花生过敏原蛋白分离纯化方法研究进展

Purification Methods of Peanut Allergens:A Review

花生中已确定的过敏原蛋白包括Ara h 1～Ara h 11 11种。本文详细介绍花生中主要过敏原蛋白(Ara h 1、Ara h 2、Ara h 3/4、Ara h 6)以及非主要过敏原蛋白(Ara h 7～Ara h 11)的分离纯化方法研究进展。花生过敏原蛋白的分离纯化方法包括硫酸铵沉淀法、柱层析法、电泳法。其中硫酸铵沉淀法主要用于粗提纯化过程,而柱层析法则主要用于花生过敏原蛋白的精制,它包括离子交换层析、凝胶过滤层析、亲和层析、疏水相互作用层析、高效液相色谱。目前离子交换层析和凝胶过滤层析在花生过敏原蛋白分离纯化中应用最为广泛,而电泳法则仅见应用于Ara h 7及油质蛋白(Ara h 10、Ara h 11)的分离纯化。

Currently,11 proteins,including Ara h 1,Ara h 2,Ara h 3,Ara h 4,Ara h 5,Ara h 6,Ara h 7,Ara h 8,Ara h 9,Ara h 10 and Ara h 11,have been identified to be peanut allergens.Recent advances in the research on separation and purification methods for major（Ara h 1,Ara h 2,Ara h 3/4 and Ara h 6） and non-major（Ara h 7-11） peanut allergens,such as ammonium sulfate precipitation,column chromatography and electrophoresis,are introduced in detail in this article.Ammonium sulfate precipitation has been generally used in the rough purification step and column chromatographic methods in the refinement step including ion exchange chromatography,gel filtration chromatography,affinity chromatography,hydrophobic interaction chromatography and high performance liquid chromatography.At present,the most extensively used method for separating and purifying peanut allergens are ion exchange chromatography and gel filtration chromatography.However,electrophoresis is only used to purify Ara h 7 and peanut oleosins（Ara h 10 and Ara h 11）.