悬铃木属花粉的纯化及免疫活性分析

目的对悬铃木属花粉变应原进行初步纯化并对其主要成分进行免疫学活性分析。方法悬铃木属花粉变应原粗制浸液经过凝胶过滤层析，收集主要蛋白质，SDS-PAGE检测各段蛋白质分子量，并用免疫印记法分析7例悬铃木属花粉过敏的支气管哮喘患者血清。结果悬铃木属花粉变应原粗制液经层析柱洗脱得两个洗脱峰，第一峰及第二峰升段富含蛋白，而第二峰降段蛋白含量甚微，收集各段进行SDS—PAGE。出现6条蛋白区带，分子量分别为7、50、35、39、22和16kd。第一峰主要含22-71kd蛋白质，峰2升段以14-16kd之蛋白质为主。对7例花粉过敏的支气管哮喘患者血清免疫印记分析可见4条sIgG反应带，蛋白分子量为50、39、22和16kd，病人血清结合百分比分别为100％、28．57％、57．14％和14．29％。结论悬铃木属花粉变应原含有6种主要蛋白成分，第一峰的50、39和22kd的蛋白质为主要致敏组分，第二峰升段的16kd蛋白质为次要致原。

Purification and immunocompetence analysis of platanus acerifolid of wild pollen allergen

Objective] To partially purify Platanus acerifolid of Wild pollen allergen and analyze the immunocompetence of its major proteins. Methods] The Platanus pollen allergens extracts were purified by gel filtration with Sephadex-G-100 and the major proteins were collected. The molecular weight (MW) of major protein was tested by sodium dodecylsulfate-polyacrylamide (SDS-PAGE) gel electrophoresis and the immunocompetence of its main proteins were analyzed by immunoblotting with sera of seven asthmatic patients allergic to Platanus pollen allergen specifically. Results] Two elution peaks were collected after gel filtration. The first peak and the ascending branch of the second peak had plenty of proteins whereas the descending branch of the second peak had little proteins. The major proteins were tested by SDS-PAGE and the results showed six protein bands with the MW were 71, 50, 35, 39, 22 and 16 kd. On SDS-PAGE, the proteins of the first peak were major in 22 to 71 kd and that of the ascending part of second peak were mainly in 14 to 16 kd. The results of immunoblotting analysis with seven asthmatic patients' sera showed 4 strip of IgG-binding components whose MW were 50, 39, 22 and 16 kd and the percentages of its combination with patient's sera were 100%, 28.57%, 57.14%, 14.29% respectively. Conclusion] The Platanus pollen allergen had six kinds of main proteins and the proteins of 50, 39, 22 kd in the first peak were the major allergenic components and that of 16 kd in ascending part of second peak was the minor allergenic components.