Løowenstein H, Markussen B, Weeke B
Int Arch Allergy Appl Immunol. 1976;51(1):48-67. doi: 10.1159/000231578.
Three major allergens of horse hair and dandruff have been isolated. The fractionation procedures involved various combinations, described in detail, of ethanol precipitation below --5degreesC, cation- and anion-exchange chromatography, and gel filtration. UV absorption, quantitative immunoelectrophoresis and RAST inhibition were used to monitor the separations. Protein impurities constituted less than 5% in all cases. The molecular weights of the isolated proteins were 1.9 X 10(4), 5.1 X 10(4) and 3.1 X 10(4) daltons, respectively. The pIs were determined as 4.1, 3.8 and 3.9, respectively. The amino acid analysis of the isolated allergens revealed large variations in their amino acid composition which might explain different reactivities in RAST experiments. The allergenic activities of the isolated antigens were determined by RAST inhibition and prick tests.
已分离出三种主要的马毛和皮屑过敏原。分级分离程序涉及在-5℃以下进行乙醇沉淀、阳离子和阴离子交换色谱以及凝胶过滤的各种详细描述的组合。使用紫外吸收、定量免疫电泳和RAST抑制来监测分离过程。在所有情况下,蛋白质杂质的含量均低于5%。分离出的蛋白质的分子量分别为1.9×10⁴、5.1×10⁴和3.1×10⁴道尔顿。等电点分别测定为4.1、3.8和3.9。对分离出的过敏原进行的氨基酸分析表明,它们的氨基酸组成存在很大差异,这可能解释了RAST实验中不同的反应性。通过RAST抑制和点刺试验测定分离出的抗原的致敏活性。