CAJ | 학술논문

目的：提取和分离纯化白花鬼针草花粉主要过敏原，并鉴定主要过敏原的致敏性。方法提取白花鬼针草花粉蛋白粗提液，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分离粗提液蛋白组分并测定其分子质量，经离子交换层析法分离纯化几类主要的蛋白，Western blotting分别检测其与花粉过敏患者血清 IgE结合情况及10位正常人阴性混合血清结合情况，对比鉴定每种主要蛋白质致敏原的致敏性。结果 SDS-PAGE 结果示：在12．5～120 ku之间广泛分布了20余条蛋白条带，其中主带7条，分子质量在35～70 ku和10～15 ku的区带蛋白含量最为丰富，余下的范围内还有约10余条次带。Westen-blotting检测示：白花鬼针草花粉变应原的致敏性强，与花粉过敏患者血清特异性 IgE结合大。离子交换层析出6类主要变应原蛋白，其分子质量分别为70、65、54、49、39及33 ku；阴性对照组 Westen-blotting在70 ku处有弱显色。结论白花鬼针草花粉的主要过敏原为70、65、54、49、39及33 ku；70 ku变应原致敏性最强。
목적：제취화분리순화백화귀침초화분주요과민원，병감정주요과민원적치민성。방법제취백화귀침초화분단백조제액，십이완기류산납-취병희선알응효전영（SDS-PAGE）분리조제액단백조분병측정기분자질량，경리자교환층석법분리순화궤류주요적단백，Western blotting분별검측기여화분과민환자혈청 IgE결합정황급10위정상인음성혼합혈청결합정황，대비감정매충주요단백질치민원적치민성。결과 SDS-PAGE 결과시：재12．5～120 ku지간엄범분포료20여조단백조대，기중주대7조，분자질량재35～70 ku화10～15 ku적구대단백함량최위봉부，여하적범위내환유약10여조차대。Westen-blotting검측시：백화귀침초화분변응원적치민성강，여화분과민환자혈청특이성 IgE결합대。리자교환층석출6류주요변응원단백，기분자질량분별위70、65、54、49、39급33 ku；음성대조조 Westen-blotting재70 ku처유약현색。결론백화귀침초화분적주요과민원위70、65、54、49、39급33 ku；70 ku변응원치민성최강。
Objective To isolate and purify the allergens from Bidens alba pollen,and to identi-fy the allergenicity of maj or allergens.Methods Pollen extracts from Bidens alba were prepared and analyzed by SDS-PAGE.The antigens were purified by ion-exchange chromatography.The binding of allergens to serum IgE was detected and the allergenicity of maj or allergens was identi-fied in patients with pollen allergy and 1 0 normal controls.Results SDS-PAGE analysis showed more than 20 protein bands at 12.5-120 ku,including 7 major bands.The 35-70 ku and 10-15 ku proteins were the most abundant proteins.Western blotting showed a strong allergenicity of Bi-dens alba pollen allergens with a extraordinary binding to serum IgE in patients with pollen aller-gy.Ion-exchange chromatography indicated that the 70,65,54,49,39 and 33 kDu proteins were the major allergens.In normal controls,Western blotting showed a weak immunoreaction at 70 kDu.Conclusion The 70,65,54,49,39 and 33 ku proteins are the major allergens and the 70 ku protein is the strongest allergen in Bidens alba pollen.