Altamirano-Bustamante A, Barnard G, Kohen F
Department of Hormone Research, Weizmann Institute of Science, Rehovot, Israel.
J Immunol Methods. 1991 Apr 8;138(1):95-101. doi: 10.1016/0022-1759(91)90068-q.
We report a novel competitive type immunoassay for oestradiol based on the idiotypic anti-idiotypic approach. This has been achieved by the production of an anti-idiotypic antibody (anti-Id) which is directed against the oestradiol binding site of the primary idiotypic antibody (Ab1). In this format the primary Ab1 was captured onto the surface of microtitre wells and oestradiol standards or serum samples were then allowed to compete with europium labelled anti-Id for the binding sites of Ab1. Fluorescence was proportional to the concentration of oestradiol over the range 0-8 ng/ml. The sensitivity of the assay was 80 +/- 20 pg/ml, whilst the intra-assay variation ranged from 3 to 10%, and the inter-assay variation from 7.3 to 15%. The results obtained by the fluorescence immunoassay correlated well with those obtained by an extraction radioimmunoassay using tritiated antigen and dextran-coated charcoal for separation of bound and free ligand (n = 60, r = 0.98). The idiotypic anti-idiotypic approach in hapten immunoassays enables antibodies to be labelled instead of haptens, and thus permits the development of robust and sensitive immunoassays.
我们报道了一种基于独特型抗独特型方法的新型雌二醇竞争性免疫测定法。这是通过产生一种抗独特型抗体（抗Id）来实现的，该抗体针对的是一级独特型抗体（Ab1）的雌二醇结合位点。在这种形式中，一级Ab1被捕获到微量滴定孔的表面，然后让雌二醇标准品或血清样品与铕标记的抗Id竞争Ab1的结合位点。荧光与0 - 8 ng/ml范围内的雌二醇浓度成正比。该测定法的灵敏度为80±20 pg/ml，批内变异范围为3%至10%，批间变异为7.3%至15%。荧光免疫测定法获得的结果与使用氚标记抗原和葡聚糖包被活性炭分离结合和游离配体的萃取放射免疫测定法获得的结果相关性良好（n = 60，r = 0.98）。半抗原免疫测定中的独特型抗独特型方法能够标记抗体而非半抗原，从而允许开发出稳健且灵敏的免疫测定法。
