Microfluidic platforms for discovery and detection of molecular biomarkers

Microfluidics has emerged as a promising platform for discovery and detection of molecular biomarkers recently. With this approach, the discovery of these biomarkers could be more efficient in time and consumes less samples and reagents. Furthermore, the entire discovery process could be automated since all the functional microfluidic devices such as micropumps and microvalves could be integrated on a single chip. Similarly, the detection of the discovered molecular biomarkers is also promising. Detection of nucleic acid biomarkers, protein biomarkers, and metabolite biomarkers has been demonstrated on microfluidic platforms recently. When compared with their large-scale counterparts, the miniature system can perform the detection of these biomarkers within less analysis time while a multiplexed detection scheme could be easily achieved. Furthermore, the entire detection process could be automated on the single chip as well. This review paper is therefore to review the recent development of microfluidic devices and systems for the discovery and detection of the molecular biomarker. Techniques for biomarker discovery, verification, and detection that have been adapted into microfluidics were first reviewed, and their advantages were highlighted. The new approach of biomarker screening based on in vitro-generated affinity reagents such as nucleic acid aptamers and peptide affinity reagents was then reviewed. Finally, in the biomarker detection section, this review placed a special emphasis on commercialized microfluidic-based diagnostics for molecular biomarkers.

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

[2]  M. Kew,et al.  Alpha-fetoprotein in primary liver cancer and other diseases. , 1974, Gut.

[3]  Gwo-Bin Lee,et al.  An integrated microfluidic system for rapid screening of alpha-fetoprotein-specific aptamers. , 2012, Biosensors & bioelectronics.

[4]  Xavier Estivill,et al.  Gene expression signatures in breast cancer distinguish phenotype characteristics, histologic subtypes, and tumor invasiveness , 2010, Cancer.

[5]  Inês Barroso,et al.  Candidate Gene Association Study in Type 2 Diabetes Indicates a Role for Genes Involved in β-Cell Function as Well as Insulin Action , 2003, PLoS biology.

[6]  P. Gaulard,et al.  Small nucleolar RNA expression profiling identifies potential prognostic markers in peripheral T-cell lymphoma. , 2012, Blood.

[7]  L. C. Gunn,et al.  Label-free quantitation of a cancer biomarker in complex media using silicon photonic microring resonators. , 2009, Analytical chemistry.

[8]  Stephen Lam,et al.  Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells. , 2007, Cancer research.

[9]  V. Ambros,et al.  The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14 , 1993, Cell.

[10]  G. Winter,et al.  Making antibodies by phage display technology. , 1994, Annual review of immunology.

[11]  Iulia M Lazar,et al.  Microfluidic chips for protein differential expression profiling , 2009, Electrophoresis.

[12]  Hui-Wen Chen,et al.  Integrated microfluidic systems for DNA analysis. , 2011, Topics in current chemistry.

[13]  E. Schiffrin,et al.  Reduction of C-reactive protein and the use of anti-hypertensives , 2007, Vascular health and risk management.

[14]  David M. Rissin,et al.  Single-Molecule enzyme-linked immunosorbent assay detects serum proteins at subfemtomolar concentrations , 2010, Nature Biotechnology.

[15]  Robert Nadon,et al.  Statistical practice in high-throughput screening data analysis , 2006, Nature Biotechnology.

[16]  W. McGuire,et al.  Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. , 1987, Science.

[17]  F. Hamdy,et al.  Very low PSA concentrations and deletions of the KLK3 gene. , 2013, Clinical chemistry.

[18]  Iulia M Lazar,et al.  Microfluidic liquid chromatography system for proteomic applications and biomarker screening. , 2006, Analytical chemistry.

[19]  Amy E. Herr,et al.  Microfluidic integration for automated targeted proteomic assays , 2012, Proceedings of the National Academy of Sciences.

[20]  Benjamin J. Raphael,et al.  Integrated Genomic Analyses of Ovarian Carcinoma , 2011, Nature.

[21]  W. Yeung,et al.  2‐D t‐ITP/CZE determination of clinical urinary proteins using a microfluidic‐chip capillary electrophoresis device , 2011, Electrophoresis.

[22]  Olivia Corso,et al.  Assessment of the Nova StatSensor whole blood point-of-care creatinine analyzer for the measurement of kidney function in screening for chronic kidney disease , 2010, Clinical chemistry and laboratory medicine.

[23]  J. Friend,et al.  Fast surface acoustic wave-matrix-assisted laser desorption ionization mass spectrometry of cell response from islets of Langerhans. , 2013, Analytical chemistry.

[24]  Rovshan G Sadygov,et al.  Large-scale database searching using tandem mass spectra: Looking up the answer in the back of the book , 2004, Nature Methods.

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

[26]  M. Ferrari,et al.  Microfluidic enrichment of small proteins from complex biological mixture on nanoporous silica chip. , 2011, Biomicrofluidics.

[27]  G. Gui,et al.  Biomarkers in the diagnosis of primary and recurrent breast cancer. , 2012, Biomarkers in medicine.

[28]  G. Meijer,et al.  Proteomics in colorectal cancer translational research: biomarker discovery for clinical applications. , 2013, Clinical biochemistry.

[29]  Kathryn A. Phillips,et al.  Diagnostics and biomarker development: priming the pipeline , 2006, Nature Reviews Drug Discovery.

[30]  L. Mazutis,et al.  Quantitative and sensitive detection of rare mutations using droplet-based microfluidics. , 2011, Lab on a chip.

[31]  H Tom Soh,et al.  Controlling the selection stringency of phage display using a microfluidic device. , 2009, Lab on a chip.

[32]  A. Poole,et al.  Protein kinase Cα: disease regulator and therapeutic target , 2010, Trends in pharmacological sciences.

[33]  R. Aebersold,et al.  Mass spectrometry-based proteomics , 2003, Nature.

[34]  M. Bowser,et al.  In vitro selection of high-affinity DNA ligands for human IgE using capillary electrophoresis. , 2004, Analytical chemistry.

[35]  I. Ges,et al.  Enzyme-coated microelectrodes to monitor lactate production in a nanoliter microfluidic cell culture device. , 2010, Biosensors & bioelectronics.

[36]  I. García-Pérez,et al.  Metabolic fingerprinting with capillary electrophoresis. , 2008, Journal of chromatography. A.

[37]  K. Strimbu,et al.  What are biomarkers? , 2010, Current opinion in HIV and AIDS.

[38]  Marc P Y Desmulliez,et al.  Lab-on-a-chip based immunosensor principles and technologies for the detection of cardiac biomarkers: a review. , 2011, Lab on a chip.

[39]  P. Brousset,et al.  Specific small nucleolar RNA expression profiles in acute leukemia , 2012, Leukemia.

[40]  R. Snell,et al.  TATA-binding protein in neurodegenerative disease , 2005, Neuroscience.

[41]  Roger N Rosenberg,et al.  Genome-wide association studies in Alzheimer disease. , 2008, Archives of neurology.

[42]  M. Wilm,et al.  Analytical properties of the nanoelectrospray ion source. , 1996, Analytical chemistry.

[43]  F. Baltazar,et al.  Monocarboxylate transporter 2 (MCT2) as putative biomarker in prostate cancer , 2013, The Prostate.

[44]  E. Petricoin,et al.  Use of proteomic patterns in serum to identify ovarian cancer , 2002, The Lancet.

[45]  Gwo-Bin Lee,et al.  A novel integrated microfluidic platform to perform fluorescence in situ hybridization for chromosomal analysis , 2013 .

[46]  Dong-Ki Lee,et al.  Selection and elution of aptamers using nanoporous sol-gel arrays with integrated microheaters. , 2009, Lab on a chip.

[47]  Paul M. Ridker,et al.  Novel risk factors for systemic atherosclerosis: a comparison of C-reactive protein, fibrinogen, homocysteine, lipoprotein(a), and standard cholesterol screening as predictors of peripheral arterial disease. , 2001 .

[48]  R. Barker,et al.  Neuropeptide Y modifies the disease course in the R6/2 transgenic model of Huntington's disease , 2010, Experimental Neurology.

[49]  Sergey N Krylov,et al.  Aptamer-facilitated biomarker discovery (AptaBiD). , 2008, Journal of the American Chemical Society.

[50]  Ming Yu,et al.  Multilayer polymer microchip capillary array electrophoresis devices with integrated on-chip labeling for high-throughput protein analysis. , 2011, Analytical chemistry.

[51]  E. Fearon Molecular Genetics of Colorectal Cancer , 1995, Annals of the New York Academy of Sciences.

[52]  Sachdev S Sidhu,et al.  Exploring Protein–Protein Interactions with Phage Display , 2003, Chembiochem : a European journal of chemical biology.

[53]  Jijun Tang,et al.  The DNA aptamers that specifically recognize ricin toxin are selected by two in vitro selection methods , 2006, Electrophoresis.

[54]  Daniel B. Martin,et al.  Circulating microRNAs as stable blood-based markers for cancer detection , 2008, Proceedings of the National Academy of Sciences.

[55]  D. DeMets,et al.  Biomarkers and surrogate endpoints: Preferred definitions and conceptual framework , 2001, Clinical pharmacology and therapeutics.

[56]  Gregor Kijanka,et al.  Protein arrays as tools for serum autoantibody marker discovery in cancer. , 2009, Journal of proteomics.

[57]  D. Levy,et al.  Multiple biomarkers for the prediction of first major cardiovascular events and death. , 2006, The New England journal of medicine.

[58]  Yue Cui,et al.  Rapid, multiplexed microfluidic phage display. , 2012, Lab on a chip.

[59]  M. Pepys,et al.  C-reactive protein and cardiovascular disease: new insights from an old molecule. , 2003, QJM : monthly journal of the Association of Physicians.

[60]  D. Mochly‐Rosen,et al.  PKC isozymes in chronic cardiac disease: possible therapeutic targets? , 2008, Annual review of pharmacology and toxicology.

[61]  P. Carroll,et al.  Microfluidic-based multiplex qRT-PCR identifies diagnostic and prognostic microRNA signatures in the sera of prostate cancer patients. , 2011, Cancer research.

[62]  L. Tiret,et al.  Genetics of Cardiovascular Diseases: From Single Mutations to the Whole Genome , 2007, Circulation.

[63]  Drew A. Hall,et al.  Quantification of Protein Interactions and Solution Transport Using High-Density GMR Sensor Arrays , 2011, Nature nanotechnology.

[64]  J. M. Castellanos,et al.  Cerebrospinal fluid neuropeptide Y in Alzheimer's disease. , 1990, European neurology.

[65]  B. Orner,et al.  Phage display screening against a set of targets to establish peptide-based sugar mimetics and molecular docking to predict binding site. , 2009, Bioorganic & medicinal chemistry.

[66]  P. Pilarski,et al.  FISH and chips: chromosomal analysis on microfluidic platforms. , 2007, IET nanobiotechnology.

[67]  Charles S Henry,et al.  Review: Microfluidic applications in metabolomics and metabolic profiling. , 2009, Analytica chimica acta.

[68]  O. Fiehn Metabolomics – the link between genotypes and phenotypes , 2004, Plant Molecular Biology.

[69]  Gwo-Bin Lee,et al.  Nucleic acid amplification using microfluidic systems. , 2013, Lab on a chip.

[70]  A. Ignatchenko,et al.  A proteome resource of ovarian cancer ascites: integrated proteomic and bioinformatic analyses to identify putative biomarkers. , 2008, Journal of proteome research.

[71]  Christina Backes,et al.  Toward the blood-borne miRNome of human diseases , 2011, Nature Methods.

[72]  D. Shangguan,et al.  Development of DNA aptamers using Cell-SELEX , 2010, Nature Protocols.

[73]  Yi Xiao,et al.  Selection of phage-displayed peptides on live adherent cells in microfluidic channels , 2011, Proceedings of the National Academy of Sciences.

[74]  Seung Soo Oh,et al.  Quantitative selection of DNA aptamers through microfluidic selection and high-throughput sequencing , 2010, Proceedings of the National Academy of Sciences.

[75]  H. Ulrich,et al.  DNA and RNA aptamers: from tools for basic research towards therapeutic applications. , 2006, Combinatorial chemistry & high throughput screening.

[76]  Pietro Amedeo Modesti,et al.  Point-of-care testing of cholesterol and triglycerides for epidemiologic studies: evaluation of the multicare-in system. , 2009, Translational research : the journal of laboratory and clinical medicine.

[77]  Yudong D. He,et al.  A Gene-Expression Signature as a Predictor of Survival in Breast Cancer , 2002 .

[78]  M. Bowser,et al.  Microfluidic selection and applications of aptamers. , 2007, Journal of separation science.

[79]  David W Piston,et al.  Microfluidic glucose stimulation reveals limited coordination of intracellular Ca2+ activity oscillations in pancreatic islets. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[80]  O. Fiehn,et al.  Metabolite profiling for plant functional genomics , 2000, Nature Biotechnology.

[81]  Sandro Carrara,et al.  NutriChip: nutrition analysis meets microfluidics. , 2013, Lab on a chip.

[82]  Ronald F Renzi,et al.  An integrated microfluidic platform for sensitive and rapid detection of biological toxins. , 2008, Lab on a chip.

[83]  Benjamin J Hindson,et al.  Droplet digital PCR measurement of HER2 copy number alteration in formalin-fixed paraffin-embedded breast carcinoma tissue. , 2013, Clinical chemistry.

[84]  E. Ruoslahti,et al.  C-end rule peptides mediate neuropilin-1-dependent cell, vascular, and tissue penetration , 2009, Proceedings of the National Academy of Sciences.

[85]  Ronald W. Davis,et al.  Quantitative Monitoring of Gene Expression Patterns with a Complementary DNA Microarray , 1995, Science.

[86]  M S Pepe,et al.  Phases of biomarker development for early detection of cancer. , 2001, Journal of the National Cancer Institute.

[87]  R. Mathies,et al.  Multichannel reverse transcription-polymerase chain reaction microdevice for rapid gene expression and biomarker analysis. , 2006, Analytical chemistry.

[88]  M. Gerstein,et al.  RNA-Seq: a revolutionary tool for transcriptomics , 2009, Nature Reviews Genetics.

[89]  Yu-Chong Tai,et al.  Microfluidic platform for liquid chromatography-tandem mass spectrometry analyses of complex peptide mixtures. , 2005, Analytical chemistry.

[90]  Arthur Weiss,et al.  ZAP-70 compared with immunoglobulin heavy-chain gene mutation status as a predictor of disease progression in chronic lymphocytic leukemia. , 2004, The New England journal of medicine.

[91]  Cécile Legallais,et al.  Metabolomics-on-a-chip and predictive systems toxicology in microfluidic bioartificial organs. , 2012, Analytical chemistry.

[92]  Gwo-Bin Lee,et al.  A DNA methylation assay for detection of ovarian cancer cells using a HpaII/MspI digestion-based PCR assay in an integrated microfluidic system , 2013 .

[93]  E Gabrielson,et al.  Aberrant methylation of p16(INK4a) is an early event in lung cancer and a potential biomarker for early diagnosis. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[94]  V. Thongboonkerd,et al.  Urinary proteome profiling using microfluidic technology on a chip. , 2007, Journal of proteome research.

[95]  S. Adelstein,et al.  Integrative Genomic Data Mining for Discovery of Potential Blood-Borne Biomarkers for Early Diagnosis of Cancer , 2008, PloS one.

[96]  Gyan Bhanot,et al.  Robust multi-tissue gene panel for cancer detection , 2010, BMC Cancer.

[97]  De-cai Yu,et al.  Circulating MicroRNAs: Potential Biomarkers for Cancer , 2011, International journal of molecular sciences.

[98]  Mark Bradley,et al.  Novel Biochip Platform for Nucleic Acid Analysis , 2012, Sensors.

[99]  O. King,et al.  Transcriptional profiling in facioscapulohumeral muscular dystrophy to identify candidate biomarkers , 2012, Proceedings of the National Academy of Sciences.

[100]  Ramesh Ramakrishnan,et al.  High Throughput Gene Expression Measurement with Real Time PCR in a Microfluidic Dynamic Array , 2008, PloS one.

[101]  D. Jed Harrison,et al.  Application of Microfluidic Devices to Proteomics Research , 2002, Molecular & Cellular Proteomics.

[102]  Sanjeeva Srivastava,et al.  Applications of protein microarrays for biomarker discovery , 2008, Proteomics. Clinical applications.

[103]  Cécile Legallais,et al.  Metabolomics-on-a-chip of hepatotoxicity induced by anticancer drug flutamide and Its active metabolite hydroxyflutamide using HepG2/C3a microfluidic biochips. , 2013, Toxicological sciences : an official journal of the Society of Toxicology.

[104]  Mehmet Koyutürk,et al.  An Integrative -omics Approach to Identify Functional Sub-Networks in Human Colorectal Cancer , 2010, PLoS Comput. Biol..

[105]  Y C Chen,et al.  A mathematical model for biopanning (affinity selection) using peptide libraries on filamentous phage. , 1995, Journal of theoretical biology.

[106]  D. Gilbert,et al.  Microfluidic extraction and stretching of chromosomal DNA from single cell nuclei for DNA fluorescence in situ hybridization , 2012, Biomedical Microdevices.

[107]  Hiroaki Suzuki,et al.  Integrated microfluidic system for the simultaneous determination of ammonia, creatinine, and urea , 2005 .

[108]  Eric Leclerc,et al.  The Current Status of Alternatives to Animal Testing and Predictive Toxicology Methods Using Liver Microfluidic Biochips , 2011, Annals of Biomedical Engineering.

[109]  E. Leclerc,et al.  Metabolomics-on-a-chip and metabolic flux analysis for label-free modeling of the internal metabolism of HepG2/C3A cells. , 2012, Molecular bioSystems.

[110]  A. Maitra,et al.  Mitochondrial DNA as a cancer biomarker. , 2005, The Journal of molecular diagnostics : JMD.

[111]  Dong-Ki Lee,et al.  A sol-gel-based microfluidics system enhances the efficiency of RNA aptamer selection. , 2011, Oligonucleotides.

[112]  D. MacGlashan,et al.  IgE antibody-specific activity in human allergic disease , 2010, Immunologic research.

[113]  David J. Mooney,et al.  Label-free biomarker detection from whole blood , 2009, 2010 10th IEEE International Conference on Solid-State and Integrated Circuit Technology.

[114]  C. Bouchard,et al.  FTO: the first gene contributing to common forms of human obesity , 2008, Obesity reviews : an official journal of the International Association for the Study of Obesity.

[115]  Sam F. Y. Li,et al.  Selection of aptamers for signal transduction proteins by capillary electrophoresis , 2010, Electrophoresis.

[116]  A. Heeger,et al.  Micromagnetic selection of aptamers in microfluidic channels , 2009, Proceedings of the National Academy of Sciences.

[117]  Elaine Holmes,et al.  Metabonomics technologies and their applications in physiological monitoring, drug safety assessment and disease diagnosis , 2004, Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals.

[118]  Charles Pound,et al.  Identification of Plasma Lipid Biomarkers for Prostate Cancer by Lipidomics and Bioinformatics , 2012, PloS one.

[119]  S. Hunt,et al.  PAI-1 in human hypertension: relation to hypertensive groups. , 2002, American journal of hypertension.

[120]  Ilaria Palchetti,et al.  Nucleic acid and peptide aptamers: fundamentals and bioanalytical aspects. , 2012, Angewandte Chemie.

[121]  M. Tewari,et al.  MicroRNA profiling: approaches and considerations , 2012, Nature Reviews Genetics.

[122]  H. Rodriguez,et al.  Mass spectrometry‐based targeted quantitative proteomics: Achieving sensitive and reproducible detection of proteins , 2012, Proteomics.

[123]  S. Ferretti,et al.  A Novel Gene Signature for Molecular Diagnosis of Human Prostate Cancer by RT-qPCR , 2008, PloS one.

[124]  S. Hanash,et al.  Emerging molecular biomarkers—blood-based strategies to detect and monitor cancer , 2011, Nature Reviews Clinical Oncology.

[125]  J. Todd,et al.  HLA-DQβ gene contributes to susceptibility and resistance to insulin-dependent diabetes mellitus , 1987, Nature.

[126]  A. Ziober,et al.  Lab‐on‐a‐chip for oral cancer screening and diagnosis , 2008, Head & neck.

[127]  Mo Chao Huang,et al.  Microsieve lab-chip device for rapid enumeration and fluorescence in situ hybridization of circulating tumor cells. , 2012, Lab on a chip.

[128]  Charles S Henry,et al.  Competitive immunoassays for simultaneous detection of metabolites and proteins using micromosaic patterning. , 2008, Analytical chemistry.

[129]  Sheng Pan,et al.  Using ‘omics’ to define pathogenesis and biomarkers of Parkinson’s disease , 2010, Expert review of neurotherapeutics.

[130]  Mehmet Toner,et al.  Clinical Microfluidics for Neutrophil Genomics and Proteomics , 2010, Nature Medicine.

[131]  T. Clackson,et al.  Making antibody fragments using phage display libraries , 1991, Nature.

[132]  Pierre Thibault,et al.  Integrated microfluidic device for mass spectrometry-based proteomics and its application to biomarker discovery programs. , 2005, Analytical chemistry.

[133]  D. Beebe,et al.  Physics and applications of microfluidics in biology. , 2002, Annual review of biomedical engineering.

[134]  Yehia Mechref,et al.  Chip-based reversed-phase liquid chromatography-mass spectrometry of permethylated N-linked glycans: a potential methodology for cancer-biomarker discovery. , 2010, Analytical chemistry.

[135]  L. Hood,et al.  Integrated barcode chips for rapid, multiplexed analysis of proteins in microliter quantities of blood , 2008, Nature Biotechnology.

[136]  Allison Doerr Mass spectrometry–based targeted proteomics , 2012, Nature Methods.

[137]  Gwo-Bin Lee,et al.  An automatic microfluidic system for rapid screening of cancer stem-like cell-specific aptamers , 2013 .

[138]  Xiaohong Fang,et al.  Aptamers generated from cell-SELEX for molecular medicine: a chemical biology approach. , 2010, Accounts of chemical research.

[139]  Daniel W. Jones,et al.  Seventh report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure. , 2003, Hypertension.

[140]  Pei Wang,et al.  A targeted proteomics–based pipeline for verification of biomarkers in plasma , 2011, Nature Biotechnology.

[141]  Christoph A. Merten,et al.  Functional single-cell hybridoma screening using droplet-based microfluidics , 2012, Proceedings of the National Academy of Sciences.

[142]  Rajesh Patel,et al.  Mutation Scanning Using MUT-MAP, a High-Throughput, Microfluidic Chip-Based, Multi-Analyte Panel , 2012, PloS one.

[143]  S. Brant,et al.  Novel candidate colorectal cancer biomarkers identified by methylation microarray-based scanning. , 2011, Endocrine-related cancer.

[144]  Hai Jiang,et al.  Microfluidic whole-blood immunoassays , 2011 .

[145]  R. Stoltenburg,et al.  SELEX--a (r)evolutionary method to generate high-affinity nucleic acid ligands. , 2007, Biomolecular engineering.

[146]  M. Bowser,et al.  Capillary electrophoresis-SELEX selection of aptamers with affinity for HIV-1 reverse transcriptase. , 2005, Analytical chemistry.

[147]  Borut Strukelj,et al.  Phage Display: Selecting Straws Instead of a Needle from a Haystack , 2011, Molecules.

[148]  E. Holland,et al.  Serum peptide profiling by magnetic particle-assisted, automated sample processing and MALDI-TOF mass spectrometry. , 2004, Analytical chemistry.

[149]  C. Dooms,et al.  Screening and early detection of lung cancer. , 2012, Annals of oncology : official journal of the European Society for Medical Oncology.

[150]  D. Spence,et al.  A microfluidic technique for monitoring bloodstream analytes indicative of C-peptide resistance in type 2 diabetes. , 2009, The Analyst.

[151]  H. Frank,et al.  Phage display: a molecular tool for the generation of antibodies--a review. , 2000, Placenta.

[152]  James F Rusling,et al.  Measurement of biomarker proteins for point-of-care early detection and monitoring of cancer. , 2010, The Analyst.

[153]  Fred S Apple,et al.  Evaluation of a point-of-care assay for cardiac markers for patients suspected of acute myocardial infarction. , 2004, Clinica chimica acta; international journal of clinical chemistry.

[154]  C. Polychronakos,et al.  Class III alleles of the variable number of tandem repeat insulin polymorphism associated with silencing of thymic insulin predispose to type 1 diabetes. , 2001, The Journal of clinical endocrinology and metabolism.

[155]  A. Jimeno,et al.  Molecular biomarkers: their increasing role in the diagnosis, characterization, and therapy guidance in pancreatic cancer , 2006, Molecular Cancer Therapeutics.

[156]  G. Hampton,et al.  DNA Methylation Profiling Defines Clinically Relevant Biological Subsets of Non–Small Cell Lung Cancer , 2012, Clinical Cancer Research.

[157]  R. Tibshirani,et al.  Classification and prediction of clinical Alzheimer's diagnosis based on plasma signaling proteins , 2007, Nature Medicine.

[158]  Julie Hardouin,et al.  HPLC-chip-mass spectrometry for protein signature identifications. , 2007, Journal of separation science.

[159]  M. McCarthy,et al.  Large-scale association studies of variants in genes encoding the pancreatic beta-cell KATP channel subunits Kir6.2 (KCNJ11) and SUR1 (ABCC8) confirm that the KCNJ11 E23K variant is associated with type 2 diabetes. , 2003, Diabetes.

[160]  Adam L. Washburn,et al.  Quantitative, label-free detection of five protein biomarkers using multiplexed arrays of silicon photonic microring resonators. , 2010, Analytical chemistry.

[161]  F. J. Wolf,et al.  THE PROPERTIES OF STREPTAVIDIN, A BIOTIN-BINDING PROTEIN PRODUCED BY STREPTOMYCETES. , 1964, Archives of biochemistry and biophysics.

[162]  Frank Larimer,et al.  Integrating 'top-down" and "bottom-up" mass spectrometric approaches for proteomic analysis of Shewanella oneidensis. , 2002, Journal of proteome research.

[163]  Seung Soo Oh,et al.  Generation of highly specific aptamers via micromagnetic selection. , 2009, Analytical chemistry.

[164]  Amy E Herr,et al.  Microfluidic immunoassays as rapid saliva-based clinical diagnostics , 2007, Proceedings of the National Academy of Sciences.

[165]  Hyon K. Choi,et al.  The genetics of hyperuricaemia and gout , 2012, Nature Reviews Rheumatology.

[166]  Gwo-Bin Lee,et al.  Integrated microfluidic system for rapid screening of CRP aptamers utilizing systematic evolution of ligands by exponential enrichment (SELEX). , 2010, Biosensors & bioelectronics.

[167]  J. Hardy,et al.  The Amyloid Hypothesis of Alzheimer ’ s Disease : Progress and Problems on the Road to Therapeutics , 2009 .

[168]  Seokheun Choi,et al.  Microfluidic-based biosensors toward point-of-care detection of nucleic acids and proteins , 2010, Microfluidics and nanofluidics.

[169]  Nicola Dalbeth,et al.  The genetic basis of hyperuricaemia and gout. , 2011, Joint, bone, spine : revue du rhumatisme.

[170]  L. Provinciali,et al.  Botulinum toxin type A for drooling in Parkinson's disease: A double‐blind, randomized, placebo‐controlled study , 2006, Movement disorders : official journal of the Movement Disorder Society.

[171]  M. Bowser,et al.  In vitro selection of aptamers with affinity for neuropeptide Y using capillary electrophoresis. , 2005, Journal of the American Chemical Society.

[172]  G. P. Smith,et al.  Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. , 1985, Science.

[173]  Gwo-Bin Lee,et al.  An automatic microfluidic system that continuously performs the systematic evolution of ligands by exponential enrichment , 2012 .

[174]  Thomas Laurell,et al.  Robotic implementation of a microchip-based protein clean-up and enrichment system for MALDI-TOF MS readout , 2006 .

[175]  Charles S. Henry,et al.  Enhanced determination of glucose by microchip electrophoresis with pulsed amperometric detection , 2004 .