Montero M T, Alonso E, Sainz T
Research Department, Hospital Ramón y Cajal, Madrid, Spain.
Allergy. 1992 Feb;47(1):26-9. doi: 10.1111/j.1398-9995.1992.tb02245.x.
Rye pollen was incubated for 30 min and proteins extracted at this time were collected as extract A (EA). The same pollen grains were resuspended in buffer and incubated for 18.5 h. Proteins extracted in this period were designated extract B (EB). Both extracts were subfractionated by DEAE ion-exchange chromatography and allergen presence in peaks detected by the dot-immunobinding technique. The results reveal that unretained proteins (peaks 1 and 2) and proteins eluted at 0.2 M NaCl from extract B contain the highest proportion of allergens. SDS-PAGE of chromatographic peaks showed that peak 2 from extract B contains a highly purified 28 kDa band. On the skin of allergic patients this band gave a stronger positive prick test than for the crude extract.
黑麦花粉孵育30分钟，此时提取的蛋白质收集为提取物A（EA）。将相同的花粉粒重悬于缓冲液中并孵育18.5小时。在此期间提取的蛋白质指定为提取物B（EB）。两种提取物都通过DEAE离子交换色谱进行亚分级分离，并通过斑点免疫结合技术检测峰中的过敏原存在情况。结果表明，未保留的蛋白质（峰1和峰2）以及从提取物B中在0.2M NaCl处洗脱的蛋白质含有最高比例的过敏原。色谱峰的SDS-PAGE显示，提取物B的峰2含有一条高度纯化的28kDa条带。在过敏患者的皮肤上，这条带比粗提取物产生更强的阳性点刺试验结果。
