长穗鱼尾葵花粉主要变应原的研究

Research on the total allergens of carvotamitis pollen

目的探讨用双向电泳对长穗鱼尾葵花粉提取液蛋白质组分进行分析的价值,并用蛋白质印迹法寻找鱼尾葵花粉主要变应原。方法提取长穗鱼尾葵花粉变应原,用双向电泳(2-DE)结合蛋白质印迹法,用长穗鱼尾葵花粉过敏阳性患者和健康对照者分别作为阳性和阴性组,获得相应的电泳图谱和免疫印迹图,对比结果以获得特异性变应原蛋白点。结果长穗鱼尾葵花粉蛋白提取液经双向电泳后,考马斯亮蓝染色,在分子量为10~170 k Da和等电点p I3.0~10.0发现约604个蛋白质点。蛋白质印迹法结果,阳性和阴性组对比,特异性变应原蛋白点数约为3个。结论可以用双向电泳联合蛋白质印迹法对长穗鱼尾葵花粉提取液蛋白质组分分析,并可进一步用于变应原致敏组分的研究。

Objective phoresis(2-DE)in protein analysis of allergen extract and to find the main allergens of carvotamitis pollen.Methods Carvotamitis pollen extract was prepared with routine method,two dimentional electrophoresis(2-DE)combined with immunoblotting assay(western-blotting).Patients with positive allergy to Carvotamitis pollen and healthy controls were used as the positive and negative groups,to obtain the 2-DE electrophoretogram and Western blotting map.Then the positive and negative results were compared to obtain specific protein spots.Results 604 soluble proteins spots were revealed in coomassie-stained gels.Most of the proteins had a molecular weight(Mr)of 10-170 k Da,and an isoelectric point(pI)value of 3.0 to 10.0.Immunoblotting of2-DE were shown and there were 3 specific antigen spots,compared with the controls.Conclusion(2-DE)is a good method in protein analysis of carvotamitis pollen allergen extract,its potential value in allergic components needs to be further studied.