辐照对蟹过敏蛋白生化性质和抗原性的影响

 【目的】研究辐照对蟹过敏蛋白生化性质和抗原性的影响。【方法】选择3、5、7、9 kGyγ射线对蟹过敏蛋白进行辐照处理,利用紫外分光光度计测定辐照后蟹过敏蛋白溶液浓度和浊度变化;荧光光度计测定辐照后蟹过敏蛋白溶液疏水性变化;SDS-聚丙烯酰胺凝胶电泳测定辐照后过敏蛋白分子量的变化;Western blot印迹和间接竞争ELISA法测定辐照后过敏蛋白抗原性变化。【结果】辐照使过敏蛋白溶液的浓度下降,浊度增加,疏水性提高,过敏蛋白条带消失,硝酸纤维素膜上无过敏条带产生,且随着辐照剂量的增加,辐照后过敏蛋白与抗体结合能力减弱。【结论】辐照使过敏蛋白的空间结构发生变化,抗原决定簇被掩盖或破坏,证明辐照技术可以有效降低蟹过敏蛋白的致敏性。

Effects of Irradiation on Biochemical Properties and Antigenicity of Crab Allergen

【Objective】 The study was conducted to evaluate the effect of γ-irradiation on the biochemical and immunological properties of an crab allergen. 【Method】 Crab protein solutions containing allergen were irradiated at 3, 5, 7 and 9 kGy. The changes of concentration, turbidity and hydrophobicity of the irradiated protein solution were determined by UV and fluorescence spectrophotometer. SDS-PAGE electrophoresis was used to analyze the change of molecular weight. Changes in antigenicity of the irradiated protein were observed by competitive indirect enzyme-linked immunosorbent assay (ELISA) and Western blotting. 【Result】 The turbidity and hydrophobicity of the crab allergen solution increased and its concentration decreased with the increase of irradiation dose. Patterns detected by the SDS-PAGE and the immunoblot showed that the band of allergy protein disappeared. Inhibition abilities of allergy protein against polyclonal IgG decreased with the increasing irradiation dose. 【Conclusion】 The conformation and epitopes on crab allergen were structurally altered by irradiation. So γ-irradiation may provide a potential method for cowering the antigenicity of crab allergen.