花生过敏原Ara h6的分离纯化及鉴定

Purification and Identification of Peanut Allergen Ara h 6

为高效分离纯化花生过敏原Ara h 6,通过脱脂、蛋白浸提、阴离子交换层析分离得到目的蛋白,并用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF/MS)及免疫印迹技术(Western blotting)对其进行鉴定。结果表明,该蛋白为花生过敏原Ara h 6,其分子质量约为15kD,纯度大于95%,得率为22.5%。该方法简单、高效,可为花生过敏的进一步研究提供实验材料。

In order to effectively isolate and purify peanut allergen Ara h 6,peanut was subjected to a series of sequential treatments,namely de-fatting,protein extraction and anion-exchange chromatographic.Sodium dodecyl sulfate polyacrylamide gel electropheresis（SDS-PAGE）,matrix-assisted laser desorption/ionization time of flight mass spectrometry（MALDI-TOF/MS） and Western blotting were used for the identification of target protein.The results indicated that the purified target protein was peanut allergen Ara h 6 with a molecular weight of 15 kD.The purity of Ara h 6 was more than 95%,and the recovery rate was 22.5%.Based on these investigations,a simple and efficient approach to purifying Ara h 6 is achieved,which will provide experimental materials for further research on peanut allergy.