Multilayer hemin/G-quadruplex wrapped gold nanoparticles as tag for ultrasensitive multiplex immunoassay by chemiluminescence imaging.
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.
State Key Laboratory of Analytical Chemistry for Life Science, Department of Chemistry, Nanjing University, Nanjing 210093, PR China.
Chen
C
Zong
Jie
J
Wu
Jie
J
Xu
Huangxian
H
Ju
Feng
F
Yan
D014408:Biomarkers, Tumor / Q000032:analysis*; D015374:Biosensing Techniques / Q000295:instrumentation; D021881:DNA, Catalytic / Q000737:chemistry*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D054856:G-Quadruplexes; D006046:Gold / Q000737:chemistry*; D006427:Hemin / Q000737:chemistry*; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D008163:Luminescent Measurements / Q000295:instrumentation*; D053768:Metal Nanoparticles / Q000032:analysis*
D014408:Biomarkers, Tumor; D021881:DNA, Catalytic; D006427:Hemin; D006046:Gold