Guzaeva T V, Komarov A M, Iurov S V, Pchelintsev S Iu, Chudinov A V, Afanas'ev S S, Zav'ialov V P
Zh Mikrobiol Epidemiol Immunobiol. 1994 Jul-Aug(4):59-63.
In this work the conditions of labeling protein A with europium ions were studied and the conjugates obtained in this study were compared with traditional peroxidase conjugates currently used in immunochemistry. The conjugates of protein A with Eu3+ chelate were obtained with the use of cyclic dianhydride of diethylenetriaminepentaacetic acid (DADETPA). Conjugation methods with the use of DADETPA was shown to permit obtaining high-quality conjugates with europium chelates. Europium-labeled protein A ensured the sensitivity of the determination of adsorbed IgG at a level of 2 ng/ml and the dynamic analytical range within 3-1,000 ng/ml, which essentially exceeded similar characteristics of peroxidase conjugates with protein A. Europium-labeled protein A was used for the detection of antibodies to Francisella tularensis in the sera of humans immunized against tularemia. The sensitivity of this assay exceeded that of the enzyme immunoassay 10- to 40-fold. A conclusion was made on the possibility of using europium labelled protein A for the determination of specific antibodies to F.tularensis. This preparation may be useful in the determination of specific antibodies in low-immune sera.
本研究探讨了用铕离子标记蛋白A的条件,并将所得结合物与免疫化学中目前使用的传统过氧化物酶结合物进行了比较。利用二乙三胺五乙酸环二酐(DADETPA)获得了蛋白A与Eu3 +螯合物的结合物。结果表明,使用DADETPA的结合方法能够获得高质量的铕螯合物结合物。铕标记的蛋白A可确保检测吸附IgG的灵敏度达到2 ng/ml,动态分析范围在3 - 1000 ng/ml之间,这基本上超过了蛋白A过氧化物酶结合物的类似特性。铕标记的蛋白A用于检测接种土拉菌疫苗的人血清中抗土拉弗朗西斯菌的抗体。该检测方法的灵敏度比酶免疫测定法高10至40倍。得出了使用铕标记蛋白A测定抗土拉弗朗西斯菌特异性抗体的可能性的结论。该制剂可能有助于低免疫血清中特异性抗体的测定。