Single-step nanoplasmonic VEGF165 aptasensor for early cancer diagnosis.

Early cancer diagnosis is very important for the prevention or mitigation of metastasis. However, effective and efficient methods are needed to improve the diagnosis and assessment of cancer. Here, we report a single-step detection method using a nanoplasmonic aptamer sensor (aptasensor), targeting a vascular endothelial growth factor-165 (VEGF(165)), a predominant biomarker of cancer angiogenesis. Our single-step detection is accomplished by (1) specific target recognition by an aptamer-target molecule interaction and (2) direct readouts of the target recognition. The readout is achieved by inactivation of surface plasmon enhancement of fluorescent probes preattached to the aptamers. Our aptasensor provides the appropriate sensitivity for clinical diagnostics with a wide range of linear detection from 25 pg/mL to 25 μg/mL (=from 1.25 pM to 1.25 μM), high specificity for VEGF(165) against PDGF-BB, osteopontin (OPN), VEGF(121), NaCl, and temporal/thermal/biological stability. In experiments with 100% serum and saliva from clinical samples, readouts of the aptasensor and an ELISA for VEGF(165) show good agreement within the limit of the ELISA kit. We envision that our developed aptasensor holds utilities for point-of-care cancer prognostics by incorporating simplicity in detection, low-cost for test, and required small sample volumes.

[1]  Luke P. Lee,et al.  Aptamer-based SERRS sensor for thrombin detection. , 2008, Nano letters.

[2]  Qiao Lin,et al.  Emerging applications of aptamers to micro- and nanoscale biosensing , 2009 .

[3]  D C Carter,et al.  Structure of serum albumin. , 1994, Advances in protein chemistry.

[4]  G. Mie.  Beiträge zur Optik trüber Medien, speziell kolloidaler Metallösungen , 1908 .

[5]  Harleen Kaur,et al.  Probing High Affinity Sequences of DNA Aptamer against VEGF165 , 2012, PloS one.

[6]  Itamar Willner,et al.  Chemiluminescent and chemiluminescence resonance energy transfer (CRET) detection of DNA, metal ions, and aptamer-substrate complexes using hemin/G-quadruplexes and CdSe/ZnS quantum dots. , 2011, Journal of the American Chemical Society.

[7]  J. Homola,et al.  Surface plasmon resonance biosensor for the detection of VEGFR-1—a protein marker of myelodysplastic syndromes , 2011, Analytical and Bioanalytical Chemistry.

[8]  Luke P. Lee,et al.  Disassembly of a core-satellite nanoassembled substrate for colorimetric biomolecular detection. , 2011, ACS nano.

[9]  Seon Joo Park,et al.  Flexible FET-type VEGF aptasensor based on nitrogen-doped graphene converted from conducting polymer. , 2012, ACS nano.

[10]  Weihong Tan,et al.  Aptamer-modified gold nanoparticles for colorimetric determination of platelet-derived growth factors and their receptors. , 2005, Analytical chemistry.

[11]  Gengfeng Zheng,et al.  Multiplexed electrical detection of cancer markers with nanowire sensor arrays , 2005, Nature Biotechnology.

[12]  L. Olsen,et al.  Aptamers embedded in polyacrylamide nanoparticles: a tool for in vivo metabolite sensing. , 2010, ACS nano.

[13]  R. Corn,et al.  Detection of protein biomarkers using RNA aptamer microarrays and enzymatically amplified surface plasmon resonance imaging. , 2007, Analytical chemistry.

[14]  A. Heeger,et al.  Label-free electronic detection of thrombin in blood serum by using an aptamer-based sensor. , 2005, Angewandte Chemie.

[15]  Anthony D. Keefe,et al.  Direct in vitro selection of a 2'-O-methyl aptamer to VEGF. , 2005, Chemistry & biology.

[16]  W. Tan,et al.  Aptamer-nanoparticle assembly for logic-based detection. , 2012, ACS applied materials & interfaces.

[17]  Yongmei Yin,et al.  A "signal-on" electrochemical aptasensor for simultaneous detection of two tumor markers. , 2012, Biosensors & bioelectronics.

[18]  Eugene S. Kim,et al.  Potent VEGF blockade causes regression of coopted vessels in a model of neuroblastoma , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[19]  Arica A Lubin,et al.  Continuous, real-time monitoring of cocaine in undiluted blood serum via a microfluidic, electrochemical aptamer-based sensor. , 2009, Journal of the American Chemical Society.

[20]  Bingling Li,et al.  SERS opens a new way in aptasensor for protein recognition with high sensitivity and selectivity. , 2007, Chemical communications.

[21]  Donhee Ham,et al.  Chip–NMR biosensor for detection and molecular analysis of cells , 2008, Nature Medicine.

[22]  Ruth Etzioni,et al.  Early detection: The case for early detection , 2003, Nature Reviews Cancer.

[23]  D. Sahani,et al.  HCC and angiogenesis: possible targets and future directions , 2011, Nature Reviews Clinical Oncology.

[24]  Eun Jeong Cho,et al.  Applications of aptamers as sensors. , 2009, Annual review of analytical chemistry.

[25]  Napoleone Ferrara,et al.  Vascular endothelial growth factor: basic science and clinical progress. , 2004, Endocrine reviews.

[26]  M. Levy,et al.  Quantum‐Dot Aptamer Beacons for the Detection of Proteins , 2005, Chembiochem : a European journal of chemical biology.

[27]  B. Chromy,et al.  Efficient maximum likelihood estimator fitting of histograms , 2010, Nature Methods.

[28]  A. Chambers,et al.  The Role of Osteopontin in Breast Cancer: Clinical and Experimental Studies , 2001, Journal of Mammary Gland Biology and Neoplasia.

[29]  A. Larsson,et al.  Serum VEGF and bFGF adds prognostic information in patients with normal platelet counts when sampled before, during and after treatment for locally advanced non-small cell lung cancer. , 2003, Lung cancer.

[30]  Sadia Afrin Khan,et al.  Multifunctional oval-shaped gold-nanoparticle-based selective detection of breast cancer cells using simple colorimetric and highly sensitive two-photon scattering assay. , 2010, ACS nano.

[31]  Rebecca Y Lai,et al.  A folding-based electrochemical aptasensor for detection of vascular endothelial growth factor in human whole blood. , 2011, Biosensors & bioelectronics.

[32]  Jian Zhang,et al.  Surface-enhanced fluorescence of fluorescein-labeled oligonucleotides capped on silver nanoparticles. , 2005, The journal of physical chemistry. B.

[33]  A. Heeger,et al.  An electronic, aptamer-based small-molecule sensor for the rapid, label-free detection of cocaine in adulterated samples and biological fluids. , 2006, Journal of the American Chemical Society.

[34]  H. Degani,et al.  Hormonal regulation of VEGF in orthotopic MCF7 human breast cancer. , 2002, Cancer research.

[35]  J. Lakowicz.  Radiative decay engineering: biophysical and biomedical applications. , 2001, Analytical biochemistry.

[36]  L. Gold,et al.  Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. , 1990, Science.

[37]  杨朝勇.  Light-switching excimer probes for rapid protein monitoring in complex biological fluids , 2005 .

[38]  S. Albani,et al.  T cell epitope-specific immune therapy for rheumatic diseases. , 2006, Arthritis and rheumatism.

[39]  D. Guyer,et al.  Pegaptanib, a targeted anti-VEGF aptamer for ocular vascular disease , 2006, Nature Reviews Drug Discovery.

[40]  Steven A Carr,et al.  Protein biomarker discovery and validation: the long and uncertain path to clinical utility , 2006, Nature Biotechnology.

[41]  S. Achilefu,et al.  Fluorescence Manipulation by Gold Nanoparticles: From Complete Quenching to Extensive Enhancement , 2011, Journal of nanobiotechnology.

[42]  R Bicknell,et al.  Vascular endothelial growth factor (VEGF) in breast cancer: comparison of plasma, serum, and tissue VEGF and microvessel density and effects of tamoxifen. , 2000, Cancer research.

[43]  J. Szostak,et al.  In vitro selection of RNA molecules that bind specific ligands , 1990, Nature.

[44]  David T. Shima,et al.  Vascular endothelial growth factor-A is a survival factor for retinal neurons and a critical neuroprotectant during the adaptive response to ischemic injury. , 2007, The American journal of pathology.

[45]  E. Schacht,et al.  Structure-activity relationships of poly(L-lysines): effects of pegylation and molecular shape on physicochemical and biological properties in gene delivery. , 2002, Journal of controlled release : official journal of the Controlled Release Society.

[46]  H. Klump.  Calorimetric studies of the interaction between DNA and poly-L-lysine. , 1976, Biophysical chemistry.

[47]  Andrey L. Rogach,et al.  Cascaded FRET in conjugated polymer/quantum dot/dye-labeled DNA complexes for DNA hybridization detection. , 2009, ACS nano.

[48]  J. Lakowicz,et al.  Fluorescence lifetime correlation spectroscopic study of fluorophore-labeled silver nanoparticles. , 2008, Analytical chemistry.

[49]  Dai-Wen Pang,et al.  Aptamer biosensor based on fluorescence resonance energy transfer from upconverting phosphors to carbon nanoparticles for thrombin detection in human plasma. , 2011, Analytical chemistry.

[50]  D. B. McIntosh,et al.  Salt Species-Dependent Electrostatic Effects on ssDNA Elasticity , 2011 .

[51]  C. Mirkin,et al.  Array-Based Electrical Detection of DNA with Nanoparticle Probes , 2002, Science.

[52]  E. Vermaas,et al.  Selection of single-stranded DNA molecules that bind and inhibit human thrombin , 1992, Nature.

[53]  S. Ell,et al.  Serum vascular endothelial growth factor in patients with head and neck squamous cell carcinoma. , 1999, Clinical otolaryngology and allied sciences.

[54]  Thomas Kelly,et al.  In vivo magnetic enrichment and multiplex photoacoustic detection of circulating tumour cells. , 2009, Nature nanotechnology.

[55]  Itamar Willner,et al.  Optical aptasensors for the analysis of the vascular endothelial growth factor (VEGF). , 2012, Analytical chemistry.

[56]  Lixia Zhao,et al.  Sensitive fluorescent sensing for DNA assay , 2010 .

[57]  Weihong Tan,et al.  Nucleic acid aptamers for biosensors and bio-analytical applications. , 2009, The Analyst.