Channel and Substrate Zone Two-Dimensional Resolution for Chemiluminescent Multiplex Immunoassay

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• 作者：Zhifeng Fu ; Zhanjun Yang ; Jinhai Tang ; Hong Liu ; Feng Yan ; Huangxian Ju
• 刊名：Analytical Chemistry
• 出版年：2007
• 出版时间：October 1, 2007
• 年：2007
• 卷：79
• 期：19
• 页码：7376 - 7382
• 全文大小：207K
• 年卷期：v.79,no.19(October 1, 2007)
• ISSN：1520-6882

文摘

A two-dimensional resolution system of channels andsubstrate zones was proposed for multiplex immunoassayperformed with a designed multichannel chemiluminescent (CL) detection device coupled with a single photomultiplier. Using carcinoma antigen 125 (CA 125),carcinoma antigen 153 (CA 153), carcinoma antigen 199(CA 199), and carcinoembryonic antigen (CEA) as twocouples of model analytes, two couples of capture antibodies were immobilized in two channels, respectively.With a sandwich format, the CL substrates for alkalinephosphatase and horseradish peroxidase were deliveredinto the channels sequentially to perform a multipleximmunoassay after the sample and tracer antibodies wereintroduced into the channels for on-line incubation. CA125, CA 153, CA 199, and CEA could be assayed in theranges of 0.50-80, 2.0-100, and 5.0-150 U/mL and1.0-70 ng/mL with limits of detection of 0.15, 0.80, and2.0 U/mL and 0.65 ng/mL at 3, respectively. The wholeassay process including regeneration of the device couldbe completed in 37 min. The proposed system showedacceptable detection and fabrication reproducibility, andthe results obtained were in acceptable agreement withthose from parallel single-analyte test of practical clinicalsera. This technique provides a new strategy for a simple,automated, and near-simultaneous multianalyte immunoassay.