Suzawa T, Ikariyama Y, Aizawa M
Department of Bioengineering, Tokyo Institute of Technology, Yokohama, Japan.
Anal Chem. 1994 Nov 15;66(22):3889-94. doi: 10.1021/ac00094a008.
A new homogeneous electroenzymatic immunoassay was developed to determine antibody concentration using glucose oxidase and ferrocene as enzymatic and electrochemical amplifier, respectively. Digoxin (Dig)-conjugated glucose oxidase (GOx) was modified with ferrocene (Fec) to form Fec-GOx-Dig conjugate. After immunocomplex formation between the Fec-GOx-Dig conjugate and the anti-digoxin antibody, the complex underwent less electrochemical reaction due to the steric hindrance of the antibody. The ferrocene multilabeled conjugate was provided for the determination of anti-digoxin antibody. Since the strategy taken here is based on a combined effect of GOx and ferrocene, i.e., enzymatic amplification by GOx and electrochemical amplification by multilabeled ferrocenes, the antibody concentration was determined in the range from 1/50 to 1/500 dilution.
开发了一种新的均相酶免疫分析法，分别使用葡萄糖氧化酶和二茂铁作为酶促和电化学放大器来测定抗体浓度。地高辛（Dig）偶联的葡萄糖氧化酶（GOx）用二茂铁（Fec）修饰，形成Fec-GOx-Dig偶联物。Fec-GOx-Dig偶联物与抗地高辛抗体形成免疫复合物后，由于抗体的空间位阻，该复合物发生的电化学反应较少。提供了二茂铁多标记偶联物用于抗地高辛抗体的测定。由于这里采用的策略基于GOx和二茂铁的联合作用，即GOx的酶促放大和多标记二茂铁的电化学放大，因此抗体浓度在1/50至1/500稀释范围内测定。
