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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
Electrochemical coding for multiplexed immunoassays of proteins.
G. Liu (2004)
Electroactive silica nanoparticles for biological labeling.
J. Wang (2006)
Ultrasensitive immunosensor for cancer biomarker proteins using gold nanoparticle film electrodes and multienzyme-particle amplification.
Vigneshwaran Mani (2009)
Electrochemical immunosensors for the simultaneous detection of two tumor markers.
M. S. Wilson (2005)
Functionalized graphene oxide as a nanocarrier in a multienzyme labeling amplification strategy for ultrasensitive electrochemical immunoassay of phosphorylated p53 (S392).
D. Du (2011)
Carbon nanotube amplification strategies for highly sensitive immunodetection of cancer biomarkers.
X. Yu (2006)
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)
Distinct pattern of p53 phosphorylation in human tumors
T. Minamoto (2001)
Functional activation of p53 via phosphorylation following DNA damage by UV but not γ radiation
Mini Kapoor (1998)
Prospects for Gold Nanorod Particles in Diagnostic and Therapeutic Applications
D. Pissuwan (2008)
Microwave triggered metal enhanced chemiluminescence: Quantitative protein determination.
M. Previte (2006)
Quantum-dot-based electrochemical immunoassay for high-throughput screening of the prostate-specific antigen.
J. Wang (2008)
Sensitive immunosensor for cancer biomarker based on dual signal amplification strategy of graphene sheets and multienzyme functionalized carbon nanospheres.
D. Du (2010)
p53 in health and disease
K. Vousden (2007)
A disposable multianalyte electrochemical immunosensor array for automated simultaneous determination of tumor markers.
J. Wu (2007)
Post-translational modification of p53 in tumorigenesis
A. Bode (2004)
Active bead-linked immunoassay on protein microarrays.
V. Morozov (2006)
Electric field-driven strategy for multiplexed detection of protein biomarkers using a disposable reagentless electrochemical immunosensor array.
J. Wu (2008)
Simultaneous immunoassay using electrochemical detection of metal ion labels.
F. J. Hayes (1994)
Electrophoresis-assisted active immunoassay.
V. Morozov (2003)
Apoferritin-templated synthesis of metal phosphate nanoparticle labels for electrochemical immunoassay.
G. Liu (2006)
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)
Regulating the p53 pathway: in vitro hypotheses, in vivo veritas
F. Toledo (2006)
Ser392 Phosphorylation Regulates the Oncogenic Function of Mutant p53
D. Yap (2004)
Dual signal amplification of glucose oxidase-functionalized nanocomposites as a trace label for ultrasensitive simultaneous multiplexed electrochemical detection of tumor markers.
G. Lai (2009)
Enzyme-functionalized silica nanoparticles as sensitive labels in biosensing.
Y. Wu (2009)
Post-translational modifications and activation of p53 by genotoxic stresses.
E. Appella (2001)
Multiplex measurement of seven tumor markers using an electrochemical protein chip.
M. S. Wilson (2006)
Ultrasensitive electrochemical immunosensor for oral cancer biomarker IL-6 using carbon nanotube forest electrodes and multilabel amplification.
R. Malhotra (2010)
Sensitive immunoassay of a biomarker tumor necrosis factor-alpha based on poly(guanine)-functionalized silica nanoparticle label.
J. Wang (2006)
Electrochemical multianalyte immunoassays using an array-based sensor.
M. S. Wilson (2006)
Rapid and reagent-saving immunoassay using innovative stirring actions of magnetic beads in microreactors in the sequential injection mode.
K. Tanaka (2005)
Flow-through multianalyte chemiluminescent immunosensing system with designed substrate zone-resolved technique for sequential detection of tumor markers.
Zhifeng Fu (2006)
Magnetic control of an electrochemical microfluidic device with an arrayed immunosensor for simultaneous multiple immunoassays.
D. Tang (2007)
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)
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Xiaoxiao Ge (2016)
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Fuyi Zhou (2017)
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Lijuan Xiao (2014)
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Jun Zhou (2012)
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Immunodiagnostics and immunosensor design (IUPAC Technical Report)
Vladimir Gubala (2014)
An innovative immunosensor for detection of tumor suppressor protein p53 in unprocessed human plasma and cancer cell lysates.
M. Hasanzadeh (2017)
Anticipating Behaviour of Advanced Materials in Healthcare
T. Arfin (2014)
Microchip device with 64-site electrode array for multiplexed immunoassay of cell surface antigens based on electrochemiluminescence resonance energy transfer.
Meisheng Wu (2012)
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)
Toward the development of smart and low cost point-of-care biosensors based on screen printed electrodes
M. U. Ahmed (2016)
Simultaneous detection of clenbuterol and ractopamine based on multiplexed competitive surface enhanced Raman scattering (SERS) immunoassay
M. Yu (2017)
Multiplex Immunosensor Arrays for Electrochemical Detection of Cancer Biomarker Proteins.
Bernard S Munge (2016)
Sensitive origami dual-analyte electrochemical immunodevice based on polyaniline/Au-paper electrode and multi-labeled 3D graphene sheets
L. Li (2014)
Gold nanorod-based electrochemical sensing of small biomolecules: A review
M. Alagiri (2017)
Electrocatalytic oxidation of tyrosines shows signal enhancement in label-free protein biosensors
Ming-yuan Wei (2012)
Fractal gold modified electrode for ultrasensitive thrombin detection.
L. Xu (2012)
Electrochemical Methods for the Analysis of Clinically Relevant Biomolecules.
Mahmoud Labib (2016)
3D ordered silver nanoshells silica photonic crystal beads for multiplex encoded SERS bioassay.
J. Li (2016)
The Advent of Salivary Breast Cancer Biomarker Detection Using Affinity Sensors
Imad Abrao Nemeir (2019)
Clinical detection of neurodegenerative blood biomarkers using graphene immunosensor
B. Li (2020)
Ultrasensitive multiplexed immunosensors for the simultaneous determination of endocrine disrupting compounds using Pt@SBA-15 as a non-enzymatic label.
H. Ma (2013)
Dual signal amplification of horseradish peroxidase functionalized nanocomposite as trace label for the electrochemical detection of carcinoembryonic antigen
Dexiang Feng (2014)
The enzyme electrocatalytic immunosensor based on functional composite nanofibers for sensitive detection of tumor suppressor protein p53
Xiaoying Wang (2015)
Electrochemical biosensing based on noble metal nanoparticles
J. Wang (2012)
Multiplexed Electrochemical Immunosensors for Clinical Biomarkers
P. Yáñez-Sedeño (2017)
Signal-on electrochemical immunoassay for APE1 using ionic liquid doped Au nanoparticle/graphene as a nanocarrier and alkaline phosphatase as enhancer.
Z. Zhong (2014)
Highly sensitive carcinoembryonic antigen detection using Ag@Au core-shell nanoparticles and dynamic light scattering
Xiangmin Miao (2014)
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)
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