NanoMonitor: a miniature electronic biosensor for glycan biomarker detection.

AIM The goal of our research is to develop an ultrasensitive diagnostic platform called 'NanoMonitor' to enable rapid label-free analysis of a highly promising class of biomarkers called glycans (oligosaccharide chains attached to proteins) with high sensitivity and selectivity. The glycosylation of fetuin - a serum protein - and extracts from a human pancreatic cancer line was analyzed to demonstrate the capabilities of the NanoMonitor. MATERIAL & METHODS The NanoMonitor device consists of a silicon chip with an array of gold electrodes forming multiple sensor sites and works on the principle of electrochemical impedance spectroscopy. Each sensor site is overlaid with a nanoporous alumina membrane that forms a high density of nanowells on top of each electrode. Lectins (proteins that bind to and recognize specific glycan structures) are conjugated to the surface of the electrode. When specific glycans from a test sample bind to lectins at the base of each nanowell, a perturbation of electrical double-layer occurs, which results in a change in the impedance. Using the lectins Sambucs nigra agglutinin (SNA) and Maackia amurensis agglutinin (MAA), subtle variations to the glycan chains of fetuin were investigated. Protein extracts from BXPC-3, a cultured human pancreatic cancer cell line were also analyzed for binding to SNA and MAA lectins. The performance of the NanoMonitor was compared to a conventional laboratory technique: lectin-based enzyme linked immunosorbent assay (ELISA). RESULTS & DISCUSSION The NanoMonitor was used to identify glycoform variants of fetuin and global differences in glycosylation of protein extracts from cultured human pancreatic cancerous versus normal cells. While results from NanoMonitor correlate very well with results from lectin-based ELISA, the NanoMonitor is rapid, completely label free, requires just 10 microl of sample, is approximately five orders of magnitude more sensitive and highly selective over a broad dynamic range of glycoprotein concentrations. CONCLUSION Based on its performance metrics, the NanoMonitor has excellent potential for development as a point-of-care handheld electronic biosensor device for routine detection of glycan biomarkers from clinical samples.

[1]  G. S. Wilson,et al.  Electrochemical biosensors: recommended definitions and classification. , 2001, Biosensors & bioelectronics.

[2]  Detection of lectin-glycan interaction using high resolution surface plasmon resonance. , 2008, The Analyst.

[3]  Allen J. Bard,et al.  Electrochemical Methods: Fundamentals and Applications , 1980 .

[4]  I. Willner,et al.  Electronic transduction of DNA sensing processes on surfaces: amplification of DNA detection and analysis of single-base mismatches by tagged liposomes. , 2001, Journal of the American Chemical Society.

[5]  M. Aubert,et al.  Restoration of alpha(1,2) fucosyltransferase activity decreases adhesive and metastatic properties of human pancreatic cancer cells. , 2000, Cancer research.

[6]  S. Prasad,et al.  Nanomonitors: Protein Biosensors for Rapid Analyte Analysis , 2008, IEEE Sensors Journal.

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

[8]  Huangxian Ju,et al.  Electrochemical and chemiluminescent immunosensors for tumor markers. , 2005, Biosensors & bioelectronics.

[9]  D. Thirstrup,et al.  Piezoelectric printing and probing of Lectin NanoProbeArrays for glycosylation analysis. , 2008, Biochemical and biophysical research communications.

[10]  T. Poon,et al.  The Potentials of Glycomics in Biomarker Discovery , 2008, Clinical Proteomics.

[11]  K. Dimock,et al.  Sialic acid tissue distribution and influenza virus tropism , 2008, Influenza and other respiratory viruses.

[12]  G. Lauc,et al.  Enzyme linked lectin assay (ELLA) for direct analysis of transferrin sialylation in serum samples. , 2007, Clinical biochemistry.

[13]  A. M. Wu,et al.  The biotin/avidin-mediated microtiter plate lectin assay with the use of chemically modified glycoprotein ligand. , 1994, Analytical biochemistry.

[14]  S. Andreescu,et al.  Studies of the binding and signaling of surface-immobilized periplasmic glucose receptors on gold nanoparticles: a glucose biosensor application. , 2008, Analytical biochemistry.

[15]  K. L. Sharpe-Timms,et al.  Glycosylation and over-expression of endometriosis-associated peritoneal haptoglobin , 2004, Glycoconjugate Journal.

[16]  Itamar Willner,et al.  Biomolecule-functionalized carbon nanotubes: applications in nanobioelectronics. , 2004, Chemphyschem : a European journal of chemical physics and physical chemistry.

[17]  Sergei Svarovsky,et al.  Label-free impedimetric detection of glycan-lectin interactions. , 2007, Analytical chemistry.

[18]  Lo Gorton,et al.  Biosensors and modern biospecific analytical techniques , 2005 .

[19]  Ana S. Viana,et al.  N-Hydroxysuccinimide-terminated self-assembled monolayers on gold for biomolecules immobilisation , 2005 .

[20]  Scott R. Kronewitter,et al.  Glycomics and disease markers. , 2009, Current opinion in chemical biology.

[21]  E. Oh,et al.  On-chip detection of protein glycosylation based on energy transfer between nanoparticles. , 2009, Biosensors & bioelectronics.

[22]  Kiyoko F. Aoki-Kinoshita,et al.  Frontiers in glycomics: Bioinformatics and biomarkers in disease An NIH White Paper prepared from discussions by the focus groups at a workshop on the NIH campus, Bethesda MD (September 11–13, 2006) , 2008, Proteomics.

[23]  Shalini Prasad,et al.  Iridium oxide nanomonitors: clinical diagnostic devices for health monitoring systems. , 2009, Biosensors & bioelectronics.

[24]  F. Lisdat,et al.  The use of electrochemical impedance spectroscopy for biosensing , 2008, Analytical and bioanalytical chemistry.

[25]  Omowunmi A Sadik,et al.  Electrochemical biosensors for monitoring the recognition of glycoprotein-lectin interactions. , 2007, Analytica chimica acta.

[26]  P. Yáñez‐Sedeño,et al.  Gold nanoparticle-based electrochemical biosensors , 2005, Analytical and bioanalytical chemistry.

[27]  B. D. Malhotra,et al.  Prospects of Nanomaterials in Biosensors , 2008 .

[28]  D. Andreescu,et al.  Chapter 7 New materials for biosensors, biochips and molecular bioelectronics , 2005 .

[29]  J. Pingarrón,et al.  Lectin-modified piezoelectric biosensors for bacteria recognition and quantification , 2008, Analytical and bioanalytical chemistry.

[30]  Joseph Wang,et al.  Electrochemical biosensors: towards point-of-care cancer diagnostics. , 2006, Biosensors & bioelectronics.

[31]  K. S. Kim,et al.  Microgravimetric lectin biosensor based on signal amplification using carbohydrate-stabilized gold nanoparticles. , 2008, Chemical communications.

[32]  T. Aastrup,et al.  Study of real-time lectin-carbohydrate interactions on the surface of a quartz crystal microbalance. , 2005, Biosensors & bioelectronics.

[33]  S. Prasad,et al.  Nanomonitors: electrical immunoassays for protein biomarker profiling. , 2008, Nanomedicine.

[34]  N. Pourmand,et al.  Label-Free Impedance Biosensors: Opportunities and Challenges. , 2007, Electroanalysis.