Zhang Chengxiao, Qi Honglan, Zhang Meining
Department of Chemistry, School of Chemistry and Materials Science, Shaanxi Normal University, Xi'an 710062, People's Republic of China.
Luminescence. 2007 Jan-Feb;22(1):53-9. doi: 10.1002/bio.926.
A highly sensitive homogeneous electrogenerated chemiluminescence (ECL) immunoassay for the determination of anti-digoxin antibody and digoxin hapten was developed employing Ru(bpy)(2)(dcbpy)NHS (bpy = 2,2'-bipyridyl; dcbpy = 2,2'-bipyridine-4,4'-dicarboxylic acid; NHS = N-hydroxysuccinimide ester) as an electrochemiluminescent label and bovine serum albumin (BSA) as a carrier protein. A digoxin hapten was indirectly heavily labelled with Ru(bpy)(2)(dcbpy)NHS through BSA to form Ru(bpy)(2)(dcbpy)NHS-BSA-digoxin conjugate. The ECL intensity of the immunocomplex of the conjugate with anti-digoxin antibody markedly decreased when the immunoreaction between Ru(bpy)(2)(dcbpy)NHS-BSA-digoxin conjugate and anti-digoxin antibody took place. Two formats, direct homogeneous immunoassay for anti-digoxin antibody and competitive immunoassay for digoxin, were developed to determine anti-digoxin antibody and digoxin, respectively. The anti-digoxin antibody concentration in the range 7.6 x 10(-8)-7.6 x 10(-6) g/mL was determined by direct homogeneous format. Digoxin hapten was determined throughout the range 4.0 x 10(-10)-1.0 x 10(-7) g/mL with a detection limit of 1.0 x 10(-10) g/mL by competitive format. The relative standard derivation for 6.0 x 10(-9) g/mL was 4.3%. The method has been applied to assaying digoxin in control human serum.
采用Ru(bpy)(2)(dcbpy)NHS（bpy = 2,2'-联吡啶；dcbpy = 2,2'-联吡啶-4,4'-二羧酸；NHS = N-羟基琥珀酰亚胺酯）作为电化学发光标记物，牛血清白蛋白（BSA）作为载体蛋白，开发了一种用于测定抗地高辛抗体和地高辛半抗原的高灵敏度均相电化学发光（ECL）免疫分析方法。地高辛半抗原通过BSA间接大量标记Ru(bpy)(2)(dcbpy)NHS，形成Ru(bpy)(2)(dcbpy)NHS-BSA-地高辛偶联物。当Ru(bpy)(2)(dcbpy)NHS-BSA-地高辛偶联物与抗地高辛抗体发生免疫反应时，该偶联物与抗地高辛抗体的免疫复合物的ECL强度显著降低。分别开发了两种分析形式，即用于抗地高辛抗体的直接均相免疫分析和用于地高辛的竞争免疫分析，以分别测定抗地高辛抗体和地高辛。通过直接均相分析形式测定抗地高辛抗体浓度范围为7.6×10(-8)-7.6×10(-6) g/mL。通过竞争分析形式在4.0×10(-10)-1.0×10(-7) g/mL范围内测定地高辛半抗原，检测限为1.0×10(-10) g/mL。6.0×10(-9) g/mL的相对标准偏差为4.3%。该方法已应用于测定正常人血清中的地高辛。
