Sakamoto H, Imai J, Iinuma K, Hayakawa S, Yanagawa Y, Ikeda I, Kurata K
Dainabot Co., Ltd. Research and Development Depertment.
Kaku Igaku. 1990 Aug;27(8):815-20.
We have developed two-site immunoradiometric assay (IRMA) for quantitation of human serum IgE, using two different anti-IgE monoclonal antibodies which recognize different epitopes on IgE molecule. We obtained 25 different monoclonal antibodies classified seven groups, and then two different monoclonal antibodies were selected for bead coater and tracer after checking the titer and specificity of each antibody. IgE IRMA we developed here showed good performance in terms of specificity, sensitivity and reproducibility. No cross-reactivity to IgG, IgM and albumin in the level of normal range was observed. The sensitivity for IgE assay was 0.6 x 10(-3) IU/tube and measurable range was 0.01-20.0 IU/tube (0.5-10(3) IU/ml). Coefficient of variations of Intra assay were 2.2-4.3% and average recovery yield was 92.5-108%. Good correlations with Phadebas IgE PRIST and Pharmacia IgE RIA were obtained i.e., y = 1.12x-0.19 (r = 0.96) and y = 0.97x-5.9 (r = 9.7), respectively. These results indicate that the assay system will contribute much to the routine diagnosis for atopic allergy disease and parasitic infections.
我们利用两种识别IgE分子上不同表位的抗IgE单克隆抗体，开发了一种用于定量人血清IgE的双位点免疫放射分析（IRMA）。我们获得了25种不同的单克隆抗体，分为七组，然后在检查每种抗体的效价和特异性后，选择两种不同的单克隆抗体分别用于包被微珠和标记物。我们在此开发的IgE IRMA在特异性、灵敏度和可重复性方面表现良好。在正常范围内未观察到与IgG、IgM和白蛋白的交叉反应。IgE检测的灵敏度为0.6×10⁻³IU/管，可测量范围为0.01 - 20.0 IU/管（0.5 - 10³IU/ml）。批内变异系数为2.2 - 4.3%，平均回收率为92.5 - 108%。与Phadebas IgE PRIST和Pharmacia IgE RIA有良好的相关性，分别为y = 1.12x - 0.19（r = 0.96）和y = 0.97x - 5.9（r = 9.7）。这些结果表明，该检测系统将对特应性过敏疾病和寄生虫感染的常规诊断有很大帮助。
