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目的对椰子花粉的变应原组分进行初步的分离、鉴定及纯化。方法提取椰子花粉粗提液,用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离椰子花粉的蛋白质组分并测定其相对分子质量,采用免疫印迹法鉴定其变应原成分,并通过离子交换层析对椰子花粉变应原进行初步分离纯化,免疫印迹进行检测。结果SDS-PAGE显示椰子花粉粗提液有10条蛋白带,其中相对分子质量(Mr)为60 000、50 000、35 000、28 000、19 000、16 000和14 000的蛋白可与椰子花粉过敏性病人血清IgE结合,且Mr50 000、16 000和14 000为主要变应原;离子交换层析结果显示主要过敏原成分主要分布在Ⅴ峰中。结论对椰子花粉变应原进行了初步的分离、鉴定和纯化,为临床椰子花粉变态反应疾病的诊断和治疗奠定了基础。
OBJECTIVE To isolate, identify and purify the allergen components of coconut pollen. Methods The crude extract of coconut pollen was extracted and the protein components of coconut pollen were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the relative molecular mass was determined. The allergen was identified by immunoblotting Components, and by ion exchange chromatography coconut pollen allergen preliminary separation and purification, Western blot detection. Results SDS-PAGE showed that there were 10 protein bands in crude coconut pollen extract. The protein with relative molecular mass (Mr) of 60 000, 50 000, 35 000, 28 000, 19 000, 16 000 and 14 000 could bind with coconut pollen Serum IgE bound to allergic patients, and Mr50 000, 16 000 and 14 000 were the major allergens; ion exchange chromatography showed that the major allergen components were mainly located in the Ⅴ peak. Conclusion The preliminary isolation, identification and purification of coconut pollen allergen has laid the foundation for the diagnosis and treatment of clinical coconut pollen allergy disease.