固体基质室温磷光免疫分析法测定人IgG  

Determination of IgG in Human Serum by Solid Substrate Room Temperature Phosphorescence-Immunoassay

作　　者：刘佳铭[1] 付艳[1] 余冰宾[1] 李隆弟[1] 

机构地区：[1]清华大学化学系,北京100084

出　　处：《高等学校化学学报》2001年第10期1645-1648,共4页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金 (批准号 :2 9775 0 13)资助

摘　　要：在聚酰胺膜 ( PM)等固体基质上 ,人免疫球蛋白 ( Ig G)能与异硫氰酸荧光素 ( FITC)标记的羊抗人抗体( GAHAb* )发生定量的特异性免疫反应 ,并保持了 FITC优良的室温磷光特性 ,据此建立了固体基质室温磷光免疫分析 ( SS-RTP-IA)测定人血清中人 Ig G的新方法 .在 0 .4μL(取样浓度为 1 .5 6～ 5 0 ng/m L)点样体积内 ,人 Ig G的含量在 0 .62 4～ 2 0 pg范围内 ,与室温磷光强度呈良好的线性关系 ,工作曲线的回归方程 ΔIp=31 .4 +4 6.2 lg[c( Ag) /pg],n=6.相关系数 r=0 .999,检出限 0 .32 pg.用于正常人血清中人 Ig G含量的测定 ,加标回收率为 96%～ 1 0 4 % ,结果满意 .On many kinds of solid substrate, such as polyamide membrane, cellulose acetate membrane etc., the specific immunological reaction between the human immunoglobulin G(IgG) and the goat anti human antibody labeled with isothiocyanate fluorescein(GAHAb *) can be carried into practice quantitatively. The antigen antibody conjugate remained excellent room temperature phosphorescence(RTP) properties of isothiocyanate fluorescein. The conditions for immunological reaction and RTP measurement were optimized. A convenient solid substrate room temperature phosphorescence immunoassay(SS RTP IA) method for determination of the IgG has been developed. The linear dynamic range of the method is 0 624-20 pg/spot(0 4 μL) and the calculated detection limit by the methodology of IUPAC is 0 32 pg/spot. The regression equation is Δ I p=31 4+46 2 lg[ c (Ag)/pg], correlation coefficient r =0 999, n =6. The method was used for the determination of IgG in normal human serum with good satisfactory results. The analytical recoveries are 96%-104% for seven replicates and the relative standard deviations are 2 6%-2 9%.

关 键 词：固体基质室温磷光免疫分析法 人IGG 羊抗人抗体 人免疫球蛋白 异硫氰酸荧光素标记 特异性免疫反应 

分 类 号：R446.6[医药卫生—诊断学] O657.39[医药卫生—临床医学]