Portnoy J, Olson I, Pacheco F, Barnes C
Children's Mercy Hospital, University of Health Sciences, Kansas City, Missouri.
Ann Allergy. 1990 Aug;65(2):109-14.
An extract of Alternaria (ALT) was passed through an affinity column containing a monoclonal antibody directed to ALT. After washing the column, a single glycoprotein was eluted using 0.1 M glycine (pH 3.0). This glycoprotein accounted for 13% of the dry weight of our ALT preparation and had a carbohydrate to protein ratio of 0.35 as compared with 3.2 for the whole ALT extract. Its molecular weight was 70 kD by SDS-PAGE and isoelectric point was 3.5 by IEF. Though individual sensitivities varied, 14/16 patients skin reactive to ALT were also reactive to this glycoprotein. Quantitative skin tests of extracts adjusted to the same dry weight per volume showed that it required about a four times greater concentration of the purified glycoprotein to give a wheal size equal to whole Alternaria. The ability to purify large amounts of this allergen with affinity chromatography should make its complete characterization possible.
将链格孢提取物（ALT）通过含有针对ALT的单克隆抗体的亲和柱。洗涤柱子后，用0.1M甘氨酸（pH 3.0）洗脱单一糖蛋白。这种糖蛋白占我们ALT制剂干重的13%，其碳水化合物与蛋白质的比例为0.35，而整个ALT提取物的该比例为3.2。通过SDS-PAGE测定其分子量为70kD，通过IEF测定等电点为3.5。尽管个体敏感性有所不同，但16例对ALT皮肤反应阳性的患者中有14例对这种糖蛋白也有反应。对每体积干重相同的提取物进行的定量皮肤试验表明，要产生与整个链格孢相同大小的风团，纯化糖蛋白所需的浓度约为其四倍。利用亲和色谱法大量纯化这种变应原的能力应使其完整表征成为可能。
