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Biosens Bioelectron. 2013 May 15;43:372-8. doi: 10.1016/j.bios.2012.12.051. Epub 2012 Dec 31.

Multilayer hemin/G-quadruplex wrapped gold nanoparticles as tag for ultrasensitive multiplex immunoassay by chemiluminescence imaging.

Author information

1
State Key Laboratory of Analytical Chemistry for Life Science, Department of Chemistry, Nanjing University, Nanjing 210093, PR China.

Abstract

A multilayer hemin/G-quadruplex DNAzyme wrapped gold nanoparticle (M-DNAzyme/AuNP) tag was designed for ultrasensitive chemiluminescence (CL) imaging. By combining with a disposable protein array, an ultrasensitive and high-throughput multiplex CL immunoassay method was proposed for simultaneous detection of four cancer biomarkers. The M-DNAzyme/AuNP tag was prepared by assembling high ratio of alkylthiol-capped signal DNA containing multiple G-quadruplex sequences to biotinylated DNA on AuNPs and then reacting with hemin to form multilayer hemin/G-quadruplex DNAzyme units. It could be bound to the biotinylated secondary antibody of sandwich immunocomplex by biotin-streptavidin conjugation to catalyze a CL reaction on a protein array, which produced strong CL emission. Under optimal conditions, the CL signals could be simultaneously collected by a charge-coupled device for ultrasensitive multiplex CL imaging of cancer biomarkers. Using α-fetoprotein, human chorionic gonadotrophin-β, carcinoma antigen 125, and carcinoembryonic antigen as model analytes, the proposed immunoassay method showed high sensitivities and wide linear ranges in a simple, cheap and high throughput way. The M-DNAzyme/AuNP as a universal signal tag as well as the protein chip could be suitable for mass production for economical, portable and multianalyte assay, showing a promising potential in application to clinic and other relative fields.

PMID:
23356995
DOI:
10.1016/j.bios.2012.12.051
[Indexed for MEDLINE]

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