Kakabakos S E, Georgiou S, Petrou P S, Christofidis I
Immunoassay Quality Control Lab., N.C.S.R. Demokritos, Institute of Radioisotopes and Radiodiagnostic Products, Aghia Paraskevi, Athens, Greece.
J Immunol Methods. 1999 Jan 1;222(1-2):183-7. doi: 10.1016/s0022-1759(98)00197-5.
A heterogeneous fluoroimmunoassay approach with measurement of the fluorescence signal directly onto the solid support is described. Fluorescein was employed as fluorescent label whereas plastic microtitration wells were used as solid carriers. The influence of the type of the plastic wells on the proposed approach was investigated. Different types of commercially available plastic microtitration wells (white-opaque, black, and transparent) were tested. The solid supports were judged on the grounds of the maximum fluorescence signal, the precision, and the sensitivity they provided in a model non-competitive assay for mouse IgG. Among the solid supports tested, the white-opaque wells provided the most promising results with respect to the parameters in question. This support was also used for the development of a competitive fluoroimmunoassay for the determination of total thyroxin in human serum samples in order to assess the validity of the proposed approach under real immunoassay conditions. The assay developed had appropriate analytical characteristics for the determination of thyroxin and the serum sample values obtained were well correlated with those determined by a commercial solid-phase radioimmunoassay.
本文描述了一种非均相荧光免疫分析方法，该方法可直接在固体支持物上测量荧光信号。采用荧光素作为荧光标记，而塑料微量滴定孔用作固体载体。研究了塑料孔类型对所提出方法的影响。测试了不同类型的市售塑料微量滴定孔（白色不透明、黑色和透明）。根据在小鼠IgG的模型非竞争性分析中提供的最大荧光信号、精密度和灵敏度对固体支持物进行评判。在所测试的固体支持物中，白色不透明孔在相关参数方面提供了最有前景的结果。该支持物还用于开发一种竞争性荧光免疫分析法，用于测定人血清样品中的总甲状腺素，以评估所提出方法在实际免疫分析条件下的有效性。所开发的分析方法具有测定甲状腺素的适当分析特性，并且获得的血清样品值与通过商业固相放射免疫分析法测定的值具有良好的相关性。
