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Highly sensitive rapid chemiluminescent immunoassay using the DNAzyme label for signal amplification.
Analyst. 2011 Oct 21; 136(20):4295-300.A

Abstract

A novel trace tag for chemiluminescent (CL) immunoassay was designed by using DNAzyme to functionalize antibody-labeled Au nanoparticles (AuNPs). The trace tag showed an excellent ability to catalyze the oxidation of luminol by hydrogen peroxide, leading to strong CL emission. By coupling the trace tag with a passive mixing accelerated immunoreaction system, a highly sensitive rapid flow-through CL immunoassay method was proposed. Using carcinoembryonic antigen (CEA) as a model analyte, the capture antibody for CEA was immobilized on paramagnetic microspheres, and DNAzyme-anti-CEA antibody functionalized AuNPs were prepared as trace tag. A three-dimensional helical glass tube kept at 37 °C in a water bath was used for passively mixing immunoreagents in a two-step sandwich immunoassay, with which each immunoreaction step could be finished within 150 s. With the help of a magnet, the immunocomplex could conveniently be separated from reactants. Compared with the horseradish peroxidase-based tag, the newly designed trace tag showed obvious signal amplification due to its strong catalytic ability and high loading ratio of DNAzyme on each AuNP. The proposed method showed a linear calibration range from 0.005 to 0.5 ng mL(-1) for CEA detection with a detection limit of 4.1 pg mL(-1) at a signal-to-noise ratio of 3 and acceptable detection reproducibility. The assay results of clinical serum samples were in acceptable agreement with the reference values. The designed immunoassay system with ultrahigh sensitivity provided a programmable and low-cost approach for high-throughput clinical application.

Authors+Show Affiliations

State Key Laboratory of Analytical Chemistry for Life Science, Department of Chemistry, Nanjing University, Nanjing, 210093, P.R. China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21881672

Citation

Wang, Chen, et al. "Highly Sensitive Rapid Chemiluminescent Immunoassay Using the DNAzyme Label for Signal Amplification." The Analyst, vol. 136, no. 20, 2011, pp. 4295-300.
Wang C, Wu J, Zong C, et al. Highly sensitive rapid chemiluminescent immunoassay using the DNAzyme label for signal amplification. Analyst. 2011;136(20):4295-300.
Wang, C., Wu, J., Zong, C., Ju, H., & Yan, F. (2011). Highly sensitive rapid chemiluminescent immunoassay using the DNAzyme label for signal amplification. The Analyst, 136(20), 4295-300. https://doi.org/10.1039/c1an15512a
Wang C, et al. Highly Sensitive Rapid Chemiluminescent Immunoassay Using the DNAzyme Label for Signal Amplification. Analyst. 2011 Oct 21;136(20):4295-300. PubMed PMID: 21881672.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Highly sensitive rapid chemiluminescent immunoassay using the DNAzyme label for signal amplification. AU - Wang,Chen, AU - Wu,Jie, AU - Zong,Chen, AU - Ju,Huangxian, AU - Yan,Feng, Y1 - 2011/09/01/ PY - 2011/9/2/entrez PY - 2011/9/2/pubmed PY - 2012/2/1/medline SP - 4295 EP - 300 JF - The Analyst JO - Analyst VL - 136 IS - 20 N2 - A novel trace tag for chemiluminescent (CL) immunoassay was designed by using DNAzyme to functionalize antibody-labeled Au nanoparticles (AuNPs). The trace tag showed an excellent ability to catalyze the oxidation of luminol by hydrogen peroxide, leading to strong CL emission. By coupling the trace tag with a passive mixing accelerated immunoreaction system, a highly sensitive rapid flow-through CL immunoassay method was proposed. Using carcinoembryonic antigen (CEA) as a model analyte, the capture antibody for CEA was immobilized on paramagnetic microspheres, and DNAzyme-anti-CEA antibody functionalized AuNPs were prepared as trace tag. A three-dimensional helical glass tube kept at 37 °C in a water bath was used for passively mixing immunoreagents in a two-step sandwich immunoassay, with which each immunoreaction step could be finished within 150 s. With the help of a magnet, the immunocomplex could conveniently be separated from reactants. Compared with the horseradish peroxidase-based tag, the newly designed trace tag showed obvious signal amplification due to its strong catalytic ability and high loading ratio of DNAzyme on each AuNP. The proposed method showed a linear calibration range from 0.005 to 0.5 ng mL(-1) for CEA detection with a detection limit of 4.1 pg mL(-1) at a signal-to-noise ratio of 3 and acceptable detection reproducibility. The assay results of clinical serum samples were in acceptable agreement with the reference values. The designed immunoassay system with ultrahigh sensitivity provided a programmable and low-cost approach for high-throughput clinical application. SN - 1364-5528 UR - https://www.unboundmedicine.com/medline/citation/21881672/Highly_sensitive_rapid_chemiluminescent_immunoassay_using_the_DNAzyme_label_for_signal_amplification_ L2 - https://doi.org/10.1039/c1an15512a DB - PRIME DP - Unbound Medicine ER -