电子束辐照对花生过敏原免疫原性及生化性质影响的研究

EFFECT OF ELECTRON BEAM IRRADIATION ON IMMUNOREACTIVITY AND BIOCHEMICAL PROPERTIES OF PEANUT ALLERGENS

研究了电子束辐照降低花生过敏原免疫原性的效果及辐照对花生生化性质的影响。选择2、4、6、8、10、15和20kGy剂量的电子束对花生过敏原液、脱脂花生粉进行辐照处理,辐照后花生主要过敏原(Ara h 1,Ara h 2,Ara h 3)蛋白分子质量的变化通过SDS-聚丙烯酰胺凝胶电泳测定,免疫原性的变化通过免疫印迹和间接竞争ELISA法测定;辐照后花生过敏原溶液的浓度、浊度及疏水性的变化通过紫外分光光度计和荧光光度计分别进行测定。结果表明,花生过敏原在溶液状态比固体状态对辐照更为敏感。当辐照剂量低于10kGy时处理的过敏原液的免疫原性稍有增强,剂量大于10kGy时免疫原性降低,剂量为20kGy时过敏原液的IC50值是未处理的11倍。辐照使过敏液的浓度和浊度随着剂量的加大而增加,疏水性与对照组相比增强,但当剂量大于15kGy时降低。电子束辐照改变了过敏原的生化性质,降低了其免疫原性,比较而言,辐照对液体状态下过敏原的影响更显著。

The effects of electron beam（EB） irradiation on allergenicity and biochemical properties of peanut protein were investigated in this study.Peanut protein extracts solution and defatted powder were irradiated at the dose of 2,4,6,8,10,15 and 20kGy.SDS-PAGE electrophoresis was used to observe the change of molecular weight.Changes in allergenicity of irradiated peanut major allergen（Ara h 1,Ara h 2,Ara h 3） were analyzed by immunoblotting and indirect enzyme-linked immunosorbent assay（CI-ELISA）.The changes of concentration,turbidity and hydrophobicity of the irradiated protein solution were determined by UV and fluorescence spectrophotometer.Results showed that allergen solution was more sensitive to irradiation than allergen in solid state.When the irradiated dose was lower than 10kGy,the immunoreactivity of allergen solution became little stronger;and when the dose higher than 10kGy,the immunoreactivity was reduced.The IC50 was 11 times than the control,after irradiation at the dose of 20kGy.The concentration and turbidity of the allergen solution increased with the increasing of irradiation dose.Hydrophobicity first went up with increasing irradiation and then decreased when the irradiation dose was above 15kGy.The biochemical properties of peanut allergen were altered and immunoreactivity was reduced by EB irradiation,the effect of electron beam irradiation on peanut allergen in solid state was more significant.