Maasch H J, Wihl J A, Schultze-Werninghaus G, Geissler W, Wahl R
Ann Allergy. 1987 Jul;59(1):29-33.
Reagents employed for RAST-inhibition need careful characterization. An allergen reference preparation was produced from carefully selected raw materials. After immunochemical and biochemical characterization, the reference preparation was used to produce allergen reference paper disks. Their quality was controlled by the amount of bound protein and serum titration, which should yield an S-shaped dose-response curve and specific binding of IgE of at least ten times more than background binding. In parallel, the biologic activity of the allergen reference preparations was established in quantitative skin prick tests in at least 20 allergic patients. From these patients, serum samples were collected and pooled after careful characterization. All three necessary reagents for RAST-inhibition (extract, disks, and serum pool) were stabilized by freeze-drying. The reproducibility of the assay procedure itself was checked by documenting the 50% inhibition value of the reference preparation, accepting only values within +/- 2 SD. By employing such reagents we could measure the activity of alum-adsorbed grass pollen extracts with sufficient precision, ie, the coefficient of variation of the mean potency of 12 batches was less than 40%.
用于RAST抑制的试剂需要仔细鉴定。一种过敏原参考制剂由精心挑选的原材料制成。经过免疫化学和生物化学鉴定后，该参考制剂被用于制作过敏原参考纸片。通过结合蛋白的量和血清滴定来控制它们的质量，这应产生一条S形剂量反应曲线，且IgE的特异性结合至少比背景结合高十倍。同时，在至少20名过敏患者的定量皮肤点刺试验中确定过敏原参考制剂的生物活性。从这些患者中收集血清样本，并在仔细鉴定后合并。用于RAST抑制的所有三种必需试剂（提取物、纸片和血清库）通过冷冻干燥进行稳定化处理。通过记录参考制剂的50%抑制值来检查测定程序本身的重现性，仅接受±2标准差范围内的值。通过使用此类试剂，我们能够足够精确地测量明矾吸附的草花粉提取物的活性，即12批产品平均效价的变异系数小于40%。
