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Multiplexed Electrochemical Immunoassay Of Phosphorylated Proteins Based On Enzyme-functionalized Gold Nanorod Labels And Electric Field-driven Acceleration.

D. Du, J. Wang, Donglai Lu, A. Dohnalkova, Y. Lin
Published 2011 · Chemistry, Medicine

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A multiplexed electrochemical immunoassay integrating enzyme amplification and electric field-driven strategy was developed for fast and sensitive quantification of phosphorylated p53 at Ser392 (phospho-p53(392)), Ser15 (phospho-p53(15)), Ser46 (phospho-p53(46)), and total p53 simultaneously. The disposable sensor array has four spatially separated working electrodes, and each of them is modified with different capture antibody, which enables simultaneous immunoassay to be conducted without cross-talk between adjacent electrodes. The enhanced sensitivity was achieved by a multienzyme amplification strategy using gold nanorods (AuNRs) as nanocarrier for coimmobilization of horseradish peroxidase (HRP) and detection antibody (Ab(2)) at a high ratio of HRP/Ab(2), which produced an amplified electrocatalytic response by the reduction of HRP oxidized thionine in the presence of hydrogen peroxide. The immunoreaction processes were accelerated by applying +0.4 V for 3 min and then -0.2 V for 1.5 min; thus, the whole sandwich immunoreactions could be completed in less than 5 min. Under optimal conditions, this method could simultaneously detect phospho-p53(392), phospho-p53(15), phospho-p53(46), and total p53 ranging from 0.01 to 20 nM, 0.05 to 20 nM, 0.1 to 50 nM, and 0.05 to 20 nM with detection limits of 5 pM, 20 pM, 30 pM, and 10 pM, respectively. Accurate determinations of these proteins in human plasma samples were demonstrated by comparison to the standard ELISA method. The disposable immunosensor array shows excellent promise for clinical screening of phosphorylated proteins and convenient point-of-care diagnostics.
This paper references
10.1021/AC049107L
Electrochemical coding for multiplexed immunoassays of proteins.
G. Liu (2004)
10.1002/SMLL.200600189
Electroactive silica nanoparticles for biological labeling.
J. Wang (2006)
10.1021/nn800863w
Ultrasensitive immunosensor for cancer biomarker proteins using gold nanoparticle film electrodes and multienzyme-particle amplification.
Vigneshwaran Mani (2009)
10.1021/AC0485278
Electrochemical immunosensors for the simultaneous detection of two tumor markers.
M. S. Wilson (2005)
10.1021/ac101715s
Functionalized graphene oxide as a nanocarrier in a multienzyme labeling amplification strategy for ultrasensitive electrochemical immunoassay of phosphorylated p53 (S392).
D. Du (2011)
10.1021/JA062117E
Carbon nanotube amplification strategies for highly sensitive immunodetection of cancer biomarkers.
X. Yu (2006)
10.1073/PNAS.95.11.6399
Ultraviolet radiation, but not gamma radiation or etoposide-induced DNA damage, results in the phosphorylation of the murine p53 protein at serine-389.
H. Lu (1998)
10.1038/sj.onc.1204458
Distinct pattern of p53 phosphorylation in human tumors
T. Minamoto (2001)
10.1073/PNAS.95.6.2834
Functional activation of p53 via phosphorylation following DNA damage by UV but not γ radiation
Mini Kapoor (1998)
10.5661/bger-25-93
Prospects for Gold Nanorod Particles in Diagnostic and Therapeutic Applications
D. Pissuwan (2008)
10.1021/AC061161+
Microwave triggered metal enhanced chemiluminescence: Quantitative protein determination.
M. Previte (2006)
10.1002/SMLL.200700459
Quantum-dot-based electrochemical immunoassay for high-throughput screening of the prostate-specific antigen.
J. Wang (2008)
10.1021/ac100036p
Sensitive immunosensor for cancer biomarker based on dual signal amplification strategy of graphene sheets and multienzyme functionalized carbon nanospheres.
D. Du (2010)
10.1038/nrm2147
p53 in health and disease
K. Vousden (2007)
10.1373/CLINCHEM.2007.086975
A disposable multianalyte electrochemical immunosensor array for automated simultaneous determination of tumor markers.
J. Wu (2007)
10.1038/nrc1455
Post-translational modification of p53 in tumorigenesis
A. Bode (2004)
10.1016/J.ACA.2005.09.068
Active bead-linked immunoassay on protein microarrays.
V. Morozov (2006)
10.1021/ac800905k
Electric field-driven strategy for multiplexed detection of protein biomarkers using a disposable reagentless electrochemical immunosensor array.
J. Wu (2008)
10.1021/AC00083A014
Simultaneous immunoassay using electrochemical detection of metal ion labels.
F. J. Hayes (1994)
10.1021/AC034733O
Electrophoresis-assisted active immunoassay.
V. Morozov (2003)
10.1002/SMLL.200600206
Apoferritin-templated synthesis of metal phosphate nanoparticle labels for electrochemical immunoassay.
G. Liu (2006)
10.1002/ADFM.200701340
Gold Nanoparticle–Colloidal Carbon Nanosphere Hybrid Material: Preparation, Characterization, and Application for an Amplified Electrochemical Immunoassay
Rongjing Cui (2008)
Gamma-radiation-induced phosphorylation of p53 on serine 15 is dose-dependent in MOLT-4 leukaemia cells.
A. Tichý (2009)
10.1038/nrc2012
Regulating the p53 pathway: in vitro hypotheses, in vivo veritas
F. Toledo (2006)
10.1158/0008-5472.CAN-1305-2
Ser392 Phosphorylation Regulates the Oncogenic Function of Mutant p53
D. Yap (2004)
10.1021/ac901996a
Dual signal amplification of glucose oxidase-functionalized nanocomposites as a trace label for ultrasensitive simultaneous multiplexed electrochemical detection of tumor markers.
G. Lai (2009)
10.1021/ac802345z
Enzyme-functionalized silica nanoparticles as sensitive labels in biosensing.
Y. Wu (2009)
10.1046/J.1432-1327.2001.02225.X
Post-translational modifications and activation of p53 by genotoxic stresses.
E. Appella (2001)
10.1021/AC060843U
Multiplex measurement of seven tumor markers using an electrochemical protein chip.
M. S. Wilson (2006)
10.1021/ac902802b
Ultrasensitive electrochemical immunosensor for oral cancer biomarker IL-6 using carbon nanotube forest electrodes and multilabel amplification.
R. Malhotra (2010)
10.1021/AC060809F
Sensitive immunoassay of a biomarker tumor necrosis factor-alpha based on poly(guanine)-functionalized silica nanoparticle label.
J. Wang (2006)
10.1021/AC0518452
Electrochemical multianalyte immunoassays using an array-based sensor.
M. S. Wilson (2006)
10.1016/j.talanta.2005.09.014
Rapid and reagent-saving immunoassay using innovative stirring actions of magnetic beads in microreactors in the sequential injection mode.
K. Tanaka (2005)
10.1021/AC0610560
Flow-through multianalyte chemiluminescent immunosensing system with designed substrate zone-resolved technique for sequential detection of tumor markers.
Zhifeng Fu (2006)
10.1373/CLINCHEM.2006.085126
Magnetic control of an electrochemical microfluidic device with an arrayed immunosensor for simultaneous multiple immunoassays.
D. Tang (2007)
10.1111/IGC.0b013e3181b70465
Expression of p53 Protein Phosphorylated at Serine 20 and Serine 392 in Malignant and Benign Ovarian Neoplasms: Correlation With Clinicopathological Parameters of Tumors
J. Bar (2009)



This paper is referenced by
10.1016/J.SNB.2014.01.112
Enzyme modified peptide nanowire as label for the fabrication of electrochemical immunosensor
Jianglin Zhang (2014)
10.1016/j.bios.2018.05.006
Dual-wavebands-resolved electrochemiluminescence multiplexing immunoassay with dichroic mirror assistant photomultiplier-tubes as detectors.
F. Zhang (2018)
10.1016/j.bios.2016.01.058
Simultaneous immunoassay of phosphorylated proteins based on apoferritin templated metallic phosphates as voltammetrically distinguishable signal reporters.
Xiaoxiao Ge (2016)
10.1016/j.aca.2017.03.038
Proximity hybridization-regulated catalytic DNA hairpin assembly for electrochemical immunoassay based on in situ DNA template-synthesized Pd nanoparticles.
Fuyi Zhou (2017)
10.1039/c3an02134c
Highly enhanced electrochemiluminescence based on pseudo triple-enzyme cascade catalysis and in situ generation of co-reactant for thrombin detection.
Lijuan Xiao (2014)
10.1016/j.bios.2012.03.025
Carbon nanospheres-promoted electrochemical immunoassay coupled with hollow platinum nanolabels for sensitivity enhancement.
Jun Zhou (2012)
10.1109/NEMS50311.2020.9265631
AC Electrothermal Flow-Enhanced, Label-Free Immunosensor for Rapid Electrochemical Sensing
J. Li (2020)
10.1515/pac-2013-1027
Immunodiagnostics and immunosensor design (IUPAC Technical Report)
Vladimir Gubala (2014)
10.1016/j.ijbiomac.2017.07.165
An innovative immunosensor for detection of tumor suppressor protein p53 in unprocessed human plasma and cancer cell lysates.
M. Hasanzadeh (2017)
10.1002/9781118774052.CH7
Anticipating Behaviour of Advanced Materials in Healthcare
T. Arfin (2014)
10.1021/ac300551e
Microchip device with 64-site electrode array for multiplexed immunoassay of cell surface antigens based on electrochemiluminescence resonance energy transfer.
Meisheng Wu (2012)
10.1016/j.ijbiomac.2020.04.271
Modification-free amperometric biosensor for the detection of wild-type p53 protein based on the in situ formation of silver nanoparticle networks for signal amplification.
Linlin Hou (2020)
10.3109/07388551.2014.992387
Toward the development of smart and low cost point-of-care biosensors based on screen printed electrodes
M. U. Ahmed (2016)
10.1039/C7NJ01394A
Simultaneous detection of clenbuterol and ractopamine based on multiplexed competitive surface enhanced Raman scattering (SERS) immunoassay
M. Yu (2017)
10.1002/elan.201600183
Multiplex Immunosensor Arrays for Electrochemical Detection of Cancer Biomarker Proteins.
Bernard S Munge (2016)
10.1016/J.ELECTACTA.2013.12.076
Sensitive origami dual-analyte electrochemical immunodevice based on polyaniline/Au-paper electrode and multi-labeled 3D graphene sheets
L. Li (2014)
10.1007/s00604-017-2418-6
Gold nanorod-based electrochemical sensing of small biomolecules: A review
M. Alagiri (2017)
10.1016/J.TRAC.2012.07.003
Electrocatalytic oxidation of tyrosines shows signal enhancement in label-free protein biosensors
Ming-yuan Wei (2012)
10.1039/c2nr30826f
Fractal gold modified electrode for ultrasensitive thrombin detection.
L. Xu (2012)
10.1021/acs.chemrev.6b00220
Electrochemical Methods for the Analysis of Clinically Relevant Biomolecules.
Mahmoud Labib (2016)
10.1039/c5cc08332j
3D ordered silver nanoshells silica photonic crystal beads for multiplex encoded SERS bioassay.
J. Li (2016)
10.3390/s19102373
The Advent of Salivary Breast Cancer Biomarker Detection Using Affinity Sensors
Imad Abrao Nemeir (2019)
10.1016/j.carbon.2020.06.048
Clinical detection of neurodegenerative blood biomarkers using graphene immunosensor
B. Li (2020)
10.1039/C3TB20932F
Ultrasensitive multiplexed immunosensors for the simultaneous determination of endocrine disrupting compounds using Pt@SBA-15 as a non-enzymatic label.
H. Ma (2013)
10.1016/J.ELECTACTA.2014.02.072
Dual signal amplification of horseradish peroxidase functionalized nanocomposite as trace label for the electrochemical detection of carcinoembryonic antigen
Dexiang Feng (2014)
10.1016/J.JELECHEM.2015.08.022
The enzyme electrocatalytic immunosensor based on functional composite nanofibers for sensitive detection of tumor suppressor protein p53
Xiaoying Wang (2015)
10.1007/s00604-011-0758-1
Electrochemical biosensing based on noble metal nanoparticles
J. Wang (2012)
10.3390/s17050965
Multiplexed Electrochemical Immunosensors for Clinical Biomarkers
P. Yáñez-Sedeño (2017)
10.1039/c4an01712a
Signal-on electrochemical immunoassay for APE1 using ionic liquid doped Au nanoparticle/graphene as a nanocarrier and alkaline phosphatase as enhancer.
Z. Zhong (2014)
10.1016/J.SNB.2013.10.016
Highly sensitive carcinoembryonic antigen detection using Ag@Au core-shell nanoparticles and dynamic light scattering
Xiangmin Miao (2014)
10.1002/ELAN.201700727
Dual Amplified Electrochemical Immunosensor for Hepatitis B Virus Surface Antigen Detection Using Hemin/G‐Quadruplex Immobilized onto Fe3O4‐AuNPs or (Hemin‐Amino‐rGO‐Au) Nanohybrid
Negar Alizadeh (2018)
10.1016/J.JELECHEM.2016.04.022
Portable detection of clenbuterol using a smartphone-based electrochemical biosensor with electric field-driven acceleration
Yanzhi Dou (2016)
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