Ovsiannikova I G, Gervazieva V B, Pershin B B
Zh Mikrobiol Epidemiol Immunobiol. 1985 Jul(7):44-8.
An EIA system for the quantitative determination of human IgE was developed. As the solid phase, polystyrene microplates sensitized with the gamma globulin fraction of sheep antiserum to human IgE were used in this system. Peroxidase conjugate with IgE was prepared with the use of periodate technique. The optimum parameters for the sorption of antibodies on polystyrene were determined: protein concentration, the pH of the buffer, temperature, the time of fixation, the working dilution of the conjugate. As the substrate of peroxidase activity, ortho-phenilenediamine solution with hydrogen peroxide was used. The result was evaluated by means of a photometer, model Titertek Multiskan MC (Flow Laboratories Ltd., Great Britain), at the wavelength 429 nm. The new EIA system proved to be specific and detected IgE in the sera under investigation at a concentration of 10(-9) g/ml. The system permitted the determination of the normal level of IgE in a group of healthy adults, this level being, on the average, 76 +/- 9 kU/1.
开发了一种用于定量测定人IgE的酶免疫分析系统。在该系统中，使用用羊抗人IgE血清的γ球蛋白组分致敏的聚苯乙烯微量板作为固相。利用高碘酸盐技术制备了与IgE结合的过氧化物酶。确定了抗体在聚苯乙烯上吸附的最佳参数：蛋白质浓度、缓冲液pH值、温度、固定时间、结合物的工作稀释度。使用含过氧化氢的邻苯二胺溶液作为过氧化物酶活性的底物。通过Titertek Multiskan MC型光度计（英国Flow Laboratories Ltd.）在429 nm波长下评估结果。新的酶免疫分析系统被证明具有特异性，能够检测出所研究血清中浓度为10(-9) g/ml的IgE。该系统可以测定一组健康成年人的IgE正常水平，该水平平均为76±9 kU/1。
