Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7138959 | Sensors and Actuators B: Chemical | 2018 | 32 Pages |
Abstract
It is crucial to detect multiple tumor markers (TMs) for cancer diagnosis. In the study, an endonuclease-linked multiplex immunoassay for TMs detection, with alpha-fetoprotein (AFP), carcino-embryonic antigen (CEA), carbohydrate antigen 199 (CA199) as models, was developed using microfluidic chip electrophoresis (MCE) for DNA analysis as platform. Firstly, three endonuclease (EDOs) labels were prepared by labeling secondary antibodies of TMs with EDOs (BamHI, PstI, EcoRI), respectively. Then, the primary antibodies of TMs were co-immobilized on the bottoms of 96-pore plate. Finally, the TMs and labels were simultaneously incubated in the pore to form various sandwich immunocomplexes. The EDOs on them can cut their corresponding DNA substrate strands into half fragments which can be separated and detected by MCE. The signal ratio (Ih/Is) of half fragments (Ih) and substrate (Is) was proportional to logarithm of TMs concentrations from 1â¯pgâ¯mLâ1 to 10â¯ngâ¯mLâ1 (Uâ¯mLâ1 for CA199) with detection limits of 0.35, 0.3â¯pgâ¯mLâ1 for AFP, CEA and 0.36â¯Uâ¯mLâ1 for CA199. The multiplex immunoassay was successfully employed in human serums. More importantly, based on the signal conversion mode, MCE for DNA analysis can be easily extended to detect many diagnosis markers and has applicable value in clinics.
Keywords
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Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Ning Gan, Linshun Xie, Kai Zhang, Yuting Cao, Futao Hu, Tianhua Li,