Fu Zhifeng, Yan Feng, Liu Hong, Yang Zhanjun, Ju Huangxian
Key Laboratory of Analytical Chemistry for Life Science (Ministry of Education of China), Department of Chemistry, Nanjing University, Nanjing, China.
Biosens Bioelectron. 2008 Feb 28;23(7):1063-9. doi: 10.1016/j.bios.2007.10.014. Epub 2007 Oct 30.
A novel flow-through immunosensing system for chemiluminescent (CL) multianalyte immunoassay was designed based on channel-resolved technique. Using alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) as model analytes, two polyethersulfone membranes modified with the corresponding capture antibodies were set in two channels of a flow cell, and then two mixtures of the sample and corresponding alkaline phosphatase labeled antibodies were introduced into the channels for on-line incubation, respectively. Upon injection of CL substrate the catalyzed CL signals from the two channels were sequentially collected with the aid of an optical shutter for CL detection of two analytes. The antibodies immobilized membranes could be regenerated for reuse. Under optimal conditions AFP and CEA could be assayed in the ranges of 5.0-150 and 0.50-80 ng/ml with detection limits of 1.5 and 0.25 ng/ml, respectively. The assay results of clinical serum samples with the proposed system were in acceptable agreement with those with the reference method in single-analyte test mode. This novel immunosensing system based on the designed channel-resolved technique provided an automated, reusable, simple, sensitive and low-cost approach for multianalyte immunoassay without using of expensive array detector.
基于通道分辨技术设计了一种用于化学发光（CL）多分析物免疫测定的新型流通式免疫传感系统。以甲胎蛋白（AFP）和癌胚抗原（CEA）作为模型分析物，将用相应捕获抗体修饰的两个聚醚砜膜设置在流通池的两个通道中，然后将样品与相应碱性磷酸酶标记抗体的两种混合物分别引入通道进行在线孵育。注入CL底物后，借助光学快门依次收集来自两个通道的催化CL信号，用于两种分析物的CL检测。固定有抗体的膜可以再生以供重复使用。在最佳条件下，AFP和CEA的检测范围分别为5.0 - 150和0.50 - 80 ng/ml，检测限分别为1.5和0.25 ng/ml。在单分析物测试模式下，该系统对临床血清样品的检测结果与参考方法的结果具有可接受的一致性。这种基于所设计的通道分辨技术的新型免疫传感系统为多分析物免疫测定提供了一种自动化、可重复使用、简单、灵敏且低成本的方法，无需使用昂贵的阵列检测器。
