Tie F, Pan A, Ru B, Wang W, Hu Y
Department of Technical Physics, Peking University, Beijing, People's Republic of China.
J Immunol Methods. 1992 Apr 27;149(1):115-20. doi: 10.1016/s0022-1759(12)80055-x.
An improved ELISA combined with linear sweep voltammetry detection of p-nitrophenol generated by an enzyme has been investigated in this study. p-nitrophenol, produced from alkaline phosphatase catalysing p-nitrophenyl phosphate, yielded an oxidative peak at 1.06 V (vs. Ag/AgCl) with a wax-impregnated tubular graphite anode. Without separation, the small three-electrode system was directly inserted in the well of an ELISA plate for detection. The detection limit for p-nitrophenol was 1 x 10(-6) M, lower than that obtained by measuring the absorbance of p-nitrophenol. The feasibility of utilizing linear sweep voltammetry as a detection scheme was demonstrated by determining metallothionein, granulocyte-colony stimulating factor and Xenopus laevis keratin using the above new system. The method was simple, reproducible and much more sensitive than traditional spectrophotometry.
本研究探索了一种改进的酶联免疫吸附测定法（ELISA），该方法结合线性扫描伏安法来检测酶产生的对硝基苯酚。碱性磷酸酶催化磷酸对硝基苯酯产生的对硝基苯酚，在涂蜡管状石墨阳极上于1.06 V（相对于Ag/AgCl）处产生一个氧化峰。无需分离，小型三电极系统直接插入酶联免疫吸附测定板的孔中进行检测。对硝基苯酚的检测限为1×10⁻⁶ M，低于通过测量对硝基苯酚吸光度所获得的检测限。通过使用上述新系统测定金属硫蛋白、粒细胞集落刺激因子和非洲爪蟾角蛋白，证明了将线性扫描伏安法用作检测方案的可行性。该方法简单、可重复，并且比传统分光光度法灵敏得多。
