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Title: An Enzyme-Free Signal Amplification Technique for Ultrasensitive Colorimetric Assay of Disease Biomarkers

Journal Article · · ACS Nano
 [1];  [2];  [3];  [2];  [2];  [4]; ORCiD logo [2]; ORCiD logo [1]
  1. Michigan Technological Univ., Houghton, MI (United States). Dept. of Chemistry
  2. Univ. of Maryland, College Park, MD (United States). Dept. of Chemistry and Biochemistry
  3. Brookhaven National Lab. (BNL), Upton, NY (United States). Condensed Matter Physics and Materials Science Dept.
  4. Brookhaven National Lab. (BNL), Upton, NY (United States). Condensed Matter Physics and Materials Science Dept.; Michigan Technological Univ., Houghton, MI (United States). Dept. of Chemical Engineering

Enzyme-based colorimetric assays have been widely used in research labs and clinical diagnosis for decades. Nevertheless, as constrained by the performance of enzymes, their detection sensitivity has not been substantially improved in recent years, which inhibits many critical applications such as early detection of cancers. In this work, we demonstrate an enzyme-free signal amplification technique, based on gold vesicles encapsulated with Pd-Ir nanoparticles as peroxidase mimics, for colorimetric assay of disease biomarkers with significantly enhanced sensitivity. This technique overcomes the intrinsic limitations of enzymes, thanks to the superior catalytic efficiency of peroxidase mimics and the efficient loading and release of these mimics. Using human prostate surface antigen as a model biomarker, we demonstrated that the enzyme-free assay could reach a limit of detection at the femtogram/mL level, which is over 103-fold lower than that of conventional enzyme-based assay when the same antibodies and similar procedure were used.

Research Organization:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Grant/Contract Number:
SC0012704
OSTI ID:
1372434
Report Number(s):
BNL--113910-2017-JA; KC0201010
Journal Information:
ACS Nano, Journal Name: ACS Nano Journal Issue: 2 Vol. 11; ISSN 1936-0851
Publisher:
American Chemical Society (ACS)Copyright Statement
Country of Publication:
United States
Language:
English

References (44)

Enhanced Gold Nanoparticle Based ELISA for a Breast Cancer Biomarker journal December 2009
Dissociation and Degradation of Thiol-Modified DNA on Gold Nanoparticles in Aqueous and Organic Solvents journal May 2011
Quantification of ricin toxin using a highly sensitive avidin/biotin enzyme-linked immunosorbent assay journal July 1988
Histochemistry as a tool in morphological analysis: a historical review journal January 2012
A First-Generation Multiplex Biomarker Analysis of Urine for the Early Detection of Prostate Cancer journal February 2008
Polymersome-based drug-delivery strategies for cancer therapeutics journal April 2015
Nanotube-Based Colorimetric Probe for Ultrasensitive Detection of Ataxia Telangiectasia Mutated Protein journal December 2011
Modified Enzyme-Linked Immunosorbent Assay Strategy Using Graphene Oxide Sheets and Gold Nanoparticles Functionalized with Different Antibody Types journal June 2013
Entropy-Driven Pattern Formation of Hybrid Vesicular Assemblies Made from Molecular and Nanoparticle Amphiphiles journal January 2014
Utilization of the biotin/avidin system to amplify the sensitivity of the enzyme-linked immunosorbent assay (ELISA) journal January 1983
Thermal Stability of DNA Functionalized Gold Nanoparticles journal November 2013
Limit of detection (LQD)/limit of quantitation (LOQ): comparison of the empirical and the statistical methods exemplified with GC-MS assays of abused drugs journal July 1994
Magnetic Bead-Based Reverse Colorimetric Immunoassay Strategy for Sensing Biomolecules journal June 2013
Magnetic silica spheres with large nanopores for nucleic acid adsorption and cellular uptake journal January 2012
A stable and highly sensitive 3,3′,5,5′-tetramethylbenzidine-based substrate reagent for enzyme-linked immunosorbent assays journal January 2000
Hydrodynamically Driven Self-Assembly of Giant Vesicles of Metal Nanoparticles for Remote-Controlled Release journal January 2013
Single-molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations journal May 2010
The Determination of Enzyme Dissociation Constants journal March 1934
Enzyme Immunoassay (EIA)/Enzyme-Linked Immunosorbent Assay (ELISA) journal December 2005
Photosensitizer-Loaded Gold Vesicles with Strong Plasmonic Coupling Effect for Imaging-Guided Photothermal/Photodynamic Therapy journal May 2013
Insights into how nucleotide supplements enhance the peroxidase-mimicking DNAzyme activity of the G-quadruplex/hemin system journal June 2012
Ultrasensitive Immunosensor for Cancer Biomarker Proteins Using Gold Nanoparticle Film Electrodes and Multienzyme-Particle Amplification journal February 2009
Crystallographic Properties of Iridium journal April 2010
Supersensitive Time-resolved Immunofluorometric Assay of Free Prostate-specific Antigen with Nanoparticle Label Technology journal July 2001
Plasmonic ELISA for the ultrasensitive detection of disease biomarkers with the naked eye journal October 2012
Paper-Based ELISA journal May 2010
Shaping binary metal nanocrystals through epitaxial seeded growth journal July 2007
Acetylcholinesterase-Catalyzed Hydrolysis Allows Ultrasensitive Detection of Pathogens with the Naked Eye journal October 2013
Nanoparticle Probes for the Detection of Cancer Biomarkers, Cells, and Tissues by Fluorescence journal April 2015
Microbead QD-ELISA: Microbead ELISA Using Biocatalytic Formation of Quantum Dots for Ultra High Sensitive Optical and Electrochemical Detection journal October 2016
Plasmonic substrates for multiplexed protein microarrays with femtomolar sensitivity and broad dynamic range journal September 2011
Use of the term "recovery" and "apparent recovery" in analytical procedures (IUPAC Recommendations 2002) journal January 2002
A synthetic strategy for carbon nanospheres impregnated with highly monodispersed metal nanoparticles journal February 2016
In vivo tumor targeting and spectroscopic detection with surface-enhanced Raman nanoparticle tags journal December 2007
Pd–Ir Core–Shell Nanocubes: A Type of Highly Efficient and Versatile Peroxidase Mimic journal September 2015
Self-Assembly of Inorganic Nanoparticle Vesicles and Tubules Driven by Tethered Linear Block Copolymers journal July 2012
Facile Synthesis of Iridium Nanocrystals with Well-Controlled Facets Using Seed-Mediated Growth journal July 2014
Versatile Immunosensor Using a Quantum Dot Coated Silica Nanosphere as a Label for Signal Amplification journal July 2010
Detection of Prostate-Specific Antigen with a Paired Surface Plasma Wave Biosensor journal April 2010
Biodegradable Gold Nanovesicles with an Ultrastrong Plasmonic Coupling Effect for Photoacoustic Imaging and Photothermal Therapy journal December 2013
Self-Assembly of Amphiphilic Plasmonic Micelle-Like Nanoparticles in Selective Solvents journal May 2013
Intrinsic peroxidase-like activity of ferromagnetic nanoparticles journal August 2007
The horseradish peroxidase-catalyzed oxidation of 3,5,3',5'-tetramethylbenzidine. Free radical and charge-transfer complex intermediates. journal April 1982
Controlled Nucleation for the Regulation of the Particle Size in Monodisperse Gold Suspensions journal January 1973

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