title: Channel and substrate zone two-dimensional resolution for chemiluminescent multiplex immunoassay.
abstract: A two-dimensional resolution system of channels and substrate zones was proposed for multiplex immunoassay performed 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 two couples of model analytes, two couples of capture antibodies were immobilized in two channels, respectively. With a sandwich format, the CL substrates for alkaline phosphatase and horseradish peroxidase were delivered into the channels sequentially to perform a multiplex immunoassay after the sample and tracer antibodies were introduced into the channels for on-line incubation. CA 125, CA 153, CA 199, and CEA could be assayed in the ranges of 0.50-80, 2.0-100, and 5.0-150 U/mL and 1.0-70 ng/mL with limits of detection of 0.15, 0.80, and 2.0 U/mL and 0.65 ng/mL at 3sigma, respectively. The whole assay process including regeneration of the device could be completed in 37 min. The proposed system showed acceptable detection and fabrication reproducibility, and the results obtained were in acceptable agreement with those from parallel single-analyte test of practical clinical sera. This technique provides a new strategy for a simple, automated, and near-simultaneous multianalyte immunoassay.
authors: Key Laboratory of Analytical Chemistry for Life Science (Ministry of Education of China), Department of Chemistry, Nanjing University, Nanjing 210093, P.R. China.
authors: Zhifeng
authors: Z
authors: Fu
authors: Zhanjun
authors: Z
authors: Yang
authors: Jinhai
authors: J
authors: Tang
authors: Hong
authors: H
authors: Liu
authors: Feng
authors: F
authors: Yan
authors: Huangxian
authors: H
authors: Ju
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis / Q000097:blood; D004563:Electrochemistry; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D049449:Luminescence
chemical_list: D014408:Biomarkers, Tumor