CAJ | 학술논문

目的：对我国蒿属花粉中常见的大籽蒿花粉进行分离、鉴定与纯化。方法：提取大籽蒿花粉粗浸液，通过聚丙烯酰胺凝胶电泳（SDS-PAGE）分离蛋白质组分，并用凝胶成像系统测定各组分的相对分子质量（Mr）；免疫印迹（Western blot）鉴定花粉主要过敏原；通过DEAE-Cellulose DE-52离子交换层析（ Ion exchange chromatography ，IEC）对大籽蒿花粉过敏原进行纯化和免疫印迹鉴定。结果：分离后得到20多种蛋白组分，其中分子量（ Mr）为62、57、38、29、25、14 kD 6个条带蛋白含量最丰富，其中Mr为62 kD和16 kD的蛋白条带与确诊的蒿属花粉过敏患者血清特异性IgE结合率最高（均＞50％），为其主要过敏原，离子交换层析纯化后可得到Mr为62、16 kD的过敏原。结论：大籽蒿花粉的主要过敏原为Mr 62 kD和16 kD的蛋白组分，离子交换层析技术可以对Mr为62 kD、16 kD的主要过敏原成分进行纯化。
목적：대아국호속화분중상견적대자호화분진행분리、감정여순화。방법：제취대자호화분조침액，통과취병희선알응효전영（SDS-PAGE）분리단백질조분，병용응효성상계통측정각조분적상대분자질량（Mr）；면역인적（Western blot）감정화분주요과민원；통과DEAE-Cellulose DE-52리자교환층석（ Ion exchange chromatography ，IEC）대대자호화분과민원진행순화화면역인적감정。결과：분리후득도20다충단백조분，기중분자량（ Mr）위62、57、38、29、25、14 kD 6개조대단백함량최봉부，기중Mr위62 kD화16 kD적단백조대여학진적호속화분과민환자혈청특이성IgE결합솔최고（균＞50％），위기주요과민원，리자교환층석순화후가득도Mr위62、16 kD적과민원。결론：대자호화분적주요과민원위Mr 62 kD화16 kD적단백조분，리자교환층석기술가이대Mr위62 kD、16 kD적주요과민원성분진행순화。
Objective:To isolate,identify and purify the Artemisia sieversiana pollen ,the mostly widespread pollen among the Artemisia pollens in China.Methods: Artemisia sieversiana extract was precipitated by saturated ammonium sulfate and then electrophoresed by SDS-PAGE.The molecular mass of each protein band was determined by gel media system.The primary allergen proteins were identified by Western blot.Allergen proteins were purified and identified by DEAE-cellulose DE-52 ion exchange chroma-tography ( IEC) and Western blot.Results: We isolated more than twenty protein bands from Artemisia sieversiana pollen extract , including the most abundant six bands whose Mr were 62 kD,57 kD,38 kD,29 kD,25 kD,14 kD espectively.The protein bands with Mr were 62 kD and 16 kD had the highest binding capacity with the specific IgE from Artemisia pollen allergic patients.The DEAE-cellulose DE-32 IEC was used to purify the primary allergen proteins with Mr 62 kD and 16 kD.Conclusion:The primary allergens of Artemisia sieversiana include the allergen proteins whose Mr are 62 kD,16 kD and the allergen of Mr 62 kD and 16 kD can be purified by chromatography.