Department of Biotechnology, College of Science, Baghdad University, Baghdad, Iraq.
Analyst. 2013 Oct 7;138(19):5647-53. doi: 10.1039/c3an00668a.
An enzyme-linked immunoassay based on dual signal transduction mechanisms has been developed for detection of ovarian cancer biomarker CA125. The immunoassay uses a nanoelectrode array (NEA) chip and absorbance methods for the dual detection. The NEA is used to confirm the optical detection of CA125 that is carried out in a high-binding 96-well plate. An alkaline phosphatase (AP) enzyme was used to label the detection antibody to allow for both the optical and electrochemical detection of CA125. Two kinds of substrates were catalyzed by the AP enzyme. para-Nitrophenylphosphate (PNPP) produces chromogenic para-nitrophenol (PNP), which can be optically detected at 405 nm. para-Aminophenylphosphate (PAPP) produces electroactive para-aminophenol (PAP), which can be detected amperometrically between -0.1 and 0.3 V. The linear ranges have been determined to be 5-1000 U mL(-1) and 5-1000 U mL(-1) for the optical and electrochemical immunoassays, respectively. The limit of detection of the optical immunoassay is 1.3 U mL(-1) and 40 U mL(-1) for the optical and electrochemical methods, respectively.
基于双重信号转导机制的酶联免疫吸附测定法已被开发用于检测卵巢癌标志物 CA125。该免疫测定法使用纳米电极阵列 (NEA) 芯片和吸光度法进行双重检测。NEA 用于确认在高结合 96 孔板中进行的 CA125 的光学检测。碱性磷酸酶 (AP) 酶被用于标记检测抗体，以允许 CA125 的光学和电化学检测。两种底物被 AP 酶催化。对硝基苯磷酸酯 (PNPP) 产生显色的对硝基苯酚 (PNP)，可在 405nm 处进行光学检测。对氨基苯磷酸酯 (PAPP) 产生电活性的对氨基酚 (PAP)，可在 -0.1 至 0.3V 之间进行安培检测。光学免疫分析的线性范围分别为 5-1000U mL(-1) 和 5-1000U mL(-1)，电化学免疫分析的线性范围分别为 5-1000U mL(-1) 和 5-1000U mL(-1)。光学免疫分析的检测限分别为 1.3U mL(-1) 和 40U mL(-1)。
