Presentation on theme: "One new technology, discover a whole new world"— Presentation transcript:
1 One new technology, discover a whole new world NanoDLSay™Nanoparticle-Enabled Dynamic Light Scattering Assay for Biological and Chemical Detection and Analysis12565 Research Parkway Suite 300Orlando, FloridaCopyright Nano Discovery Inc. 2012
2 One new technology, discover a whole new world NanoDLSay™: a new analytical technique that uses particle size change for signal transductionGold nanoparticles (AuNPs) are used as the optical probe in NanoDLSay™
3 A protein complex(> 20 nm)Immunoglobulin G(IgG) (~7-10 nm)Viruses (~10s nm)Liu X, Dai Q, Austin L, Coutts J, Knowles G, J, Chen H, H Q. A One-step homogeneous immunoassay for cancer biomarker detection using gold nanoparticle probes coupled with dynamic light scattering. J. Am. Chem. Soc. 2008; 130:2. Dai Q, Liu X, Coutts J, Austin L, Q. A one-step highly sensitive method for DNA detection using dynamic light scattering. J. Am. Chem. Soc. 2008; 130:When analytes are bound to the nanoparticles, this will cause various changes to the particle size…Citrate-protected AuNP ( nm)AuNP immunoprobeD » 120 nmAuNP immune probe bound with a small protein monomerD » nmAuNP immunoprobe bound with a large protein complexD > nmYAuNPs bound with metal ion targets through metal-chelating ligandsAuNPs bound with small chemical targets through coordinative ligand interactionsUnmodified AuNPD = 100 nmD >> 100 nm2+A small chemical(< a few nm)A protein monomer (~5-20 nm)
4 NanoDLSay™ Procedure One-step homogenous solution assay Required sample volume: 1-5 µLResults obtain in a few minutes
6 Publications Protein detection and protein cancer biomarker research: Liu X, Dai Q, Austin L, Coutts J, Knowles G, Zou J, Chen H, Huo Q. A One-step homogeneous immunoassay for cancer biomarker detection using gold nanoparticle probes coupled with dynamic light scattering. J. Am. Chem. Soc. 2008; 130:Liu X, Huo, Q. A washing-free and amplification-free one-step homogeneous assay for protein detection using gold nanoparticle probes and dynamic light scattering. J. Immun. Method 2009; 349:Jans H, Liu X, Austin L, Maes G, Huo Q. Dynamic light scattering as a powerful tool for gold nanoparticle bioconjugation and biomolecular binding study. Anal. Chem. 2009; 81:Austin L, Liu X, Huo Q. An immunoassay for monoclonal antibody isotyping and quality analysis using gold nanoparticles and dynamic light scattering. American Biotechnology Laboratory 2010; 28: 8,Bogdanovic J, Colon J, Baker C, Huo Q. A label-free nanoparticle aggregation assay for protein complex/aggregate detection and analysis. Anal. Biochem. 2010; 45:Huo Q. Protein complexes/aggregates as potential cancer biomarkers revealed by a nanoparticle aggregation assay. Colloids Surfaces B 2010; 78:Huo Q, Colon J, Codero A, Bogdanovic J, Baker CH, Goodison S, Pensky MY. A facile nanoparticle immunoassay for cancer biomarker discovery. J. Nanobiotechnology 2011; 9:20 (open access).Jaganathan S, Yue P, Paladino DC, Bogdanovic J, Huo Q, Turkson J. A functional nuclear epidermal growth factor receptor, Src and Stat3 heteromeric complex in pancreatic cancer cells. PLoS One 2011, 6(5):e19605 (open access).Chun C, Joo J, Kwon D, Kim CS, Cha HJ, Chung MS, Jeon S. A facile and sensitive immunoassay for the detection of alpha-fetoprotein using gold-coated magnetic nanoparticle clusters and dynamic light scattering. Chem. Comm. 2011, 47, Wang, X.; Ramström, O.; Yan, M. Dynamic light scattering as an efficient tool to study glyconanoparticle- lectin interactions. Analyst 2011, 136,
7 Publications DNA detection: Virus detection: Small chemical detection: Dai Q, Liu X, Coutts J, Austin L, Huo Q. A one-step highly sensitive method for DNA detection using dynamic light scattering. J. Am. Chem. Soc. 2008; 130:Gao D, Sheng Z, Han H. An ultrasensitive method for the detection of gene fragment from transgenics using label-free gold nanoparticle probe and dynamic light scattering. Anal. Chim Acta 2011; 696:1- 5. Miao XM, Xiong C, Wei WW, Ling LS, Shuai XT. Dynamic light scattering based sequence-specific recognition of double-stranded DNA with oligonucleotide functionalized gold nanoparticles. Chem. Eur. J. 2011, 17, Pylaev TE, Khanadeev VA, Khlebtsov BN, Dykman LA, Bogatyrev VA, Khlebtsov NG. Colorimetric and dynamic light scattering detection of DNA sequences by using positively charged gold nanospheres: a comparative study with gold nanorods. Nanotechnology 2011; 22: (11pp)Virus detection:Driskell JD, Jones CA, Tompkins SM, Tripp RA. One-step assay for detecting influenza virus using dynamic light scattering and gold nanoparticles. Analyst 2011; 136:Small chemical detection:Yang X, Huang J, Wang Q, Wang K, Yang L, Huo X. A one-step sensitive dynamic light scattering method for adenosine detection using split aptamer fragments. Anal Method 2011; 3: Dasary SSR, Senapati D, Singh AK, Anjaneyulu Y, Yu H, Ray PC. Highly sensitive and selective dynamic light scattering assay for TNT detection using p-ATP attached gold nanoparticles. ACS Appl. Mater. Interface 2010; 2:
8 Publications Toxic metal ion detection: Kalluri JR, Arbneshi T, Khan SA, Nelly A, Candice P, Varisli B, Washington M, McAfee S, Robinson B, Banerjee S, Singh AK, Senapati D, Ray PC. Use of gold nanoparticles in a simple colorimetric and ultrasensitive dynamic light scattering assay: selective detection of arsenic in groundwater. Angew. Chem. Int. Ed. 2009; 48:Beqa L, Singh AK, Khan SA, Senapati D, Arumugam SR, Ray PC. Gold nanoparticle-based simple colorimetric and ultrasensitive dynamic light scattering assay for the selective detection of Pb(II) from paints, plastics, and water samples. ACS Appl. Mater. Interfaces 2011; 3:Miao X, Ling L, Shua X. Ultrasensitive detection of lead(II) with DNAzyme and gold nanoparticles probes by using a dynamic light scattering technique. Chem. Comm. 2011; 47:Zhang L, Yao Y, Shan J, Li H. Lead (II) ion detection in surface water with pM sensitivity using aza- crown-ether-modified silver nanoparticles via dynamic light scattering. Nanotechnology 2011; 22: (8pp)Miao X, Ling L.; Shuai X. Detection of Pb2+ at attomole levels by using dynamic light scattering and unmodified gold nanoparticles. Anal. Biochem. 2012, 421,Xiong C, Ling L. Label-free, sensitive detection of Hg(II) with gold nanoparticles by using dynamic light scattering technique. Talanta 2012, 89,
9 Analytical Performance AnalytesSensitivityDynamic RangeProteinsHigh pg/mL to low ng/mL range2-3 orders of magnitudeDNAs30 fM(5 orders of magnitude more sensitive than SPR and fluorescence techniques)> 5 orders of magnitudeViruses< 100 TCID50/mL(1-2 orders of magnitude more sensitive than commercial diagnostic kits)Toxic metal ionsArsenics: 10 ppt (WHO acceptable limit: 10 ppb)Lead: 100 ppt (2 orders of magnitude below the EPA standard limit)Small molecules7 nM(5 orders of magnitude more sensitive than the colorimetric method)> 4 orders of magnitudeExplosive chemicals100 pMNotes:(1) ng-nanogram; fg-femtogram; fM-femtomolar; pM-picomolar; nM-nanomolar; ppb-parts per billion; ppt-parts per trillion; TCID50- 50% tissue culture infective dose. (2) All data were taken from published papers. Refer to the list of publications for more information. (3) WHO: World Health Organization; EPA: Environmental Protection Agency.
10 Analytical Performance Comparison of NanoDLSay™ with other methods for DNA detectionLabelMethodDetection limitAuNPColorimetric1 × 10-8 mol/LAu chipSurface plasmon resonance1 × 10-9 mol/LAu/polyaniline nantubeElectrochemical impedance spectroscopy3 × mol/LQuantum dotsAnodic stripping voltammetry5 × mol/LZnS and CdSe quantum dotsFluorescence2 × 10-9 mol/LNanoDLSay™Dynamic light scattering3 × mol/LRef: Gao D, Sheng Z, Han H. An ultrasensitive method for the detection of gene fragment from transgenics using label-free gold nanoparticle probe and dynamic light scattering. Anal. Chim Acta 2011; 696:1-5.
11 Advantages NanoDLSay™ Extremely simple Ultra-high sensitivity Excellent reproducibilityHomogeneous solution assayExtremely easy to learn and useData is easy to collect and interpretRequire small volume of sample (<5 µL)No special sample preparation is requiredApplicable to a wide range of sample matricesUltra-low cost of consumables to conduct the assayResults are obtained in minutes instead of hours or daysReveals new molecular information that cannot be or has not been detected by any other existing techniquesNanoDLSay™
12 One new technology, discover a whole new world NanoDLSay™: The most comprehensive and powerful tool available for protein detection and analysis
13 X Limitations of traditional ELISA Comparisons A B Individual protein monomerProtein complexProtein aggregatesABantibodyXSuitable for detecting individual proteinsNot suitable when protein complexes are present
14 NanoDLSay™Average particle size increase (nm)Incubation time (min) = 2D of analyte0 min30 min123Kinetic binding study: monitor the particle size change continuously during the assayDetermine the “size” of the target analyte at a saturated binding levelDetermine if a target protein is a monomer, complex, or aggregatesLabel-free detection: no need to label the target proteinsDetection of protein complexes and aggregates from real biological samples
15 Particle size change upon antibody addition NanoDLSay™A two-step assay for protein complex detection and binding partner analysisAverage particle size increase (nm)Incubation time (min)Step 2: Antibody screeningStep 1: Catch the targetParticle size change upon antibody additionc ~ 2DBinding partnersNot binding partners0 min60 min
16 Co-Immunoprecipitation NanoDLSay™: a two-step processCompleted in ~30 minCo-Immunoprecipitation (Co-IP):Multiple steps (approximately steps)Takes hours to days to complete
17 Non-specific interactions Co-IP and NanoDLSay™:Which one is more specific?Centrifuge in Co-IP process increases non- specific interactionsNon-specific proteins are “caught” by the particles due to increased particle concentration towards the end of centrifugeThis problem is not present in NanoDLSay™NanoDLSay™ is more specific than Co-IPCentrifuge
18 Label-free detectionNanoDLSay™ for label-free protein oligomer and aggregate detection from real biological samplesYYNanoDLSay™Other techniquesFluorescence techniques require the labeling of target proteinsSize exclusion chromatography (SEC) and analytical ultracentrifuge (AU) are only suitable for pure protein solution studySEC may underestimate the aggregation level due to solvent elution; while AU may overestimate the aggregation level due to centrifuge-induced enrichmentLabel freeDetects protein oligomers and aggregates directly from real biological samplesDoes not change the aggregation status of the sample during the assayHigh to ultra-high sensitivity
19 Product & ServicesNDS1200: A new dynamic light scattering instrument designed for NanoDLSay™Automatic measurement of 12 samplesFast analysis time: 10-20s per sample40 µL assay solution is used for the measurementLow-cost, disposable min-glass tubes with caps are used as sample containers.No cross-contamination between samplesHigh throughput capability: samples/hourThe hardware is maintenance-freeNo special housing environment is required
20 Product & ServicesNanoDLSay™ software: A software designed for convenient, flexible and high throughput analysis
21 Order Information NDS1200 NDS-Kit100 Dynamic light scattering instrument for conducting NanoDLSay™NDS-Kit100Assay kit including disposable sample cells and other consumablesPlease Contact Us to Request a Quote:12565 Research Parkway Suite 300Orlando, FL 32826Phone:Or visit online:www. nanodiscoveryinc.comNotesPatent application pending on NanoDLSay™ technology and NDS1200 system: PCT/US09/ and PCT/US11/21002Nano Discovery Inc. has the exclusive license in the world to practice and commercialize NanoDLSay™ technology
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