Leuvering J H, Thal P J, van der Waart M, Schuurs A H
J Immunoassay. 1980;1(1):77-91. doi: 10.1080/01971528008055777.
We describe the use of inorganic (metal) colloidal particles as a label for immunoassays. Dose-response curves for human placental lactogen (HPL) and human chorionic gonadotrophin (HCG) were obtained with sandwich immunoassays, using conjugates consisting of antibody-coated colloidal gold or silver particles. Several techniques were used to measure the amount of bound conjugate, viz. colorimetry and carbon rod atomic absorption spectrophotometry (CRAAS). At higher antigen concentrations the results of the assay could be read by the naked eye. Using gold particles as label and CRAAS as detection method, we found a detection limit for a sandwich HPL sol particle immunoassay (SPIA) of 1,4 pmol/l, which was equal to that of an optimalized competitive radioimmunoassay. When using a colorimeter the detection limit for HPL of this SPIA was 5,4 pmol/l, which was superior to that of a corresponding sandwich enzyme-immunoassay (EIA). HPL and HCG were also simultaneously determined, using microtitration plates, coated with a mixture of anti-HPL and anti-HCG, and a mixture of silver particle anti-HPL conjugate and gold particle anti-HCG conjugate. CRAAS was used to measure the bound amount of silver and gold conjugate. This simultaneous assay requires more work in order to obtain better sensitivities.
我们描述了使用无机(金属)胶体颗粒作为免疫测定的标记物。采用包被抗体的胶体金或银颗粒缀合物,通过夹心免疫测定法获得了人胎盘催乳素(HPL)和人绒毛膜促性腺激素(HCG)的剂量反应曲线。使用了几种技术来测量结合缀合物的量,即比色法和碳棒原子吸收分光光度法(CRAAS)。在较高抗原浓度下,测定结果可用肉眼读取。以金颗粒作为标记物,CRAAS作为检测方法,我们发现夹心HPL溶胶颗粒免疫测定法(SPIA)的检测限为1.4 pmol/l,这与优化后的竞争性放射免疫测定法相当。当使用比色计时,该SPIA对HPL的检测限为5.4 pmol/l,优于相应的夹心酶免疫测定法(EIA)。还使用包被有抗HPL和抗HCG混合物的微量滴定板以及银颗粒抗HPL缀合物和金颗粒抗HCG缀合物的混合物同时测定HPL和HCG。使用CRAAS测量银和金缀合物的结合量。这种同时测定需要更多的工作以获得更高的灵敏度。