研制了基于抗体包被化学镀纳米金(AuNPs)和[Cu(bpy)2(ONO)]NO3配合物(CuL)共固定修饰玻碳电极(GCE)的安培免疫传感器,并用于血清中癌胚抗原(CEA)的检测。首先将GCE电极表面氧化形成羧基,进而键合上乙二胺。将此胺化电极浸泡在CuL和化学镀金溶液后,CuL可通过π-π堆积作用吸附到GCE表面,并在电极表面还原成30~50nm的纳米金层(GCE|CuL/AuNPs)。将上述电极浸泡在CEA抗体(anti CEA)溶液中,利用AuNPs固定anti CEA,并通过辣根过氧化物酶(HRP)封闭剩余的AuNPs位点,由此构建了一类快速检测CEA的无试剂安培免疫传感器(GCE|CuL-AuNPs/anti CEA-HRP)。其中CuL作为电子媒介体对过氧化脲(CP)有催化还原作用,而且HRP可增强这种作用。当该传感器在37℃下,含CEA的pH6.5PBS溶液中温育30min后,随着温育液中CEA浓度的增加,电极表面形成的免疫复合物也增加,导致CuL对CP的催化电流下降。电流下降百分比1%与CEA浓度在0.1~80ng/mL之间成线性关系,检测限为0.052ng/mL(3σ)。由于采用化学镀法可以方便地在GCE表面制备纳米金膜进而包被抗体,并通过π-π堆积作用吸附CuL作为电子媒介体,故该免疫电极制备简单;采用可催化CP还原的HRP封闭AuNPs层多余位点,大大提高了电极灵敏度,有望用血清中痕量CEA分析。 Ampere immunosensor based on co-immobilization of glassy carbon electrode (GCE) with electroless gold nanoparticles (AuNPs) and [Cu (bpy) 2 (ONO)] NO3 complexes (CuL) Antigen (CEA) detection. First, the surface of the GCE electrode is oxidized to form a carboxyl group, and then the ethylenediamine is bonded. After soaking the amination electrode in CuL and electroless gold plating solution, CuL can be adsorbed onto the surface of GCE by π-π stacking and reduced to 30-50 nm (GCE | CuL / AuNPs) on the surface of electrode. The above electrodes were soaked in anti CEA solution, immobilized with anti-CEA using AuNPs, and the remaining AuNPs sites were blocked by horseradish peroxidase (HRP). Thus, a reagent-less Ampere Immunosensor (GCE | CuL-AuNPs / anti CEA-HRP). Among them, CuL acts as an electron mediator for the catalytic reduction of carbamide peroxide (CP), and HRP enhances this effect. When the sensor was incubated for 30 min at 37 ° C in CEA-containing pH 6.5 PBS solution, the immunocomplex formed on the surface of the electrode also increased as the concentration of CEA in the incubation solution increased, resulting in a decrease in the catalytic current of CuL to CP. The percentage decrease of 1% of current was linear with the concentration of CEA in the range of 0.1 ~ 80ng / mL with the detection limit of 0.052ng / mL (3σ). Due to the electroless plating method, the gold nanoparticles can be easily prepared on the surface of GCE to coat the antibody, and the CuL can be adsorbed by the π-π deposition as an electronic medium. Therefore, the preparation of the immunopotentiating electrode is simple; the HRP encapsulating AuNPs Layer extra sites, greatly increased the sensitivity of the electrode, is expected to use trace serum CEA analysis.