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NanoDLSay Nanoparticle-Enabled Dynamic Light Scattering Assay for Biological and Chemical Detection and Analysis 12565 Research Parkway Suite 300 Orlando,

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Presentation on theme: "NanoDLSay Nanoparticle-Enabled Dynamic Light Scattering Assay for Biological and Chemical Detection and Analysis 12565 Research Parkway Suite 300 Orlando,"— Presentation transcript:

1 NanoDLSay Nanoparticle-Enabled Dynamic Light Scattering Assay for Biological and Chemical Detection and Analysis Research Parkway Suite 300 Orlando, Florida One new technology, discover a whole new world Copyright Nano Discovery Inc. 2012

2 NanoDLSay : a new analytical technique that uses particle size change for signal transduction Gold nanoparticles ( AuNPs ) are used as the optical probe in NanoDLSay One new technology, discover a whole new world

3 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: 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: Citrate-protected AuNP ( nm) A protein monomer (~5-20 nm) A small chemical (< a few nm) AuNP immunoprobe D 120 nm AuNP immune probe bound with a small protein monomer D nm AuNP immunoprobe bound with a large protein complex D > nm Y Y Y Y Y Y Y Y Y Y Y Y Y Y AuNPs bound with metal ion targets through metal-chelating ligands AuNPs bound with small chemical targets through coordinative ligand interactions Unmodified AuNP D = 100 nm D >> 100 nm 2+ Y Y Y Y Y Y Y A protein complex (> 20 nm) Immunoglobulin G (IgG) (~7-10 nm) Viruses (~10s nm ) When analytes are bound to the nanoparticles, this will cause various changes to the particle size…

4 NanoDLSay Procedure One-step homogenous solution assay Required sample volume: 1-5 µL Results obtain in a few minutes

5 NanoDLSay Proteins DNAs RNAs Viruses Small chemicals Toxic metal ions APPLICATIONS

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 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: DNA 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. C olorimetric and dynamic light scattering detection of DNA sequences by using positively charged gold nanospheres: a comparative study with gold nanorods. Nanotechnology 2011; 22: (11pp)

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 Range ProteinsHigh pg/mL to low ng/mL range2-3 orders of magnitude DNAs30 fM (5 orders of magnitude more sensitive than SPR and fluorescence techniques) > 5 orders of magnitude Viruses< 100 TCID 50 /mL (1-2 orders of magnitude more sensitive than commercial diagnostic kits) 2-3 orders of magnitude Toxic metal ionsArsenics: 10 ppt (WHO acceptable limit: 10 ppb) Lead: 100 ppt (2 orders of magnitude below the EPA standard limit) 2-3 orders of magnitude Small molecules7 nM (5 orders of magnitude more sensitive than the colorimetric method) > 4 orders of magnitude Explosive chemicals100 pM2-3 orders of magnitude Notes: (1) ng-nanogram; fg-femtogram; fM-femtomolar; pM-picomolar; nM-nanomolar; ppb-parts per billion; ppt-parts per trillion; TCID % 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 Ref: 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. LabelMethodDetection limit AuNPColorimetric1 × mol/L Au chipSurface plasmon resonance1 × mol/L Au/polyaniline nantube Electrochemical impedance spectroscopy 3 × mol/L Quantum dotsAnodic stripping voltammetry5 × mol/L ZnS and CdSe quantum dots Fluorescence2 × mol/L NanoDLSayDynamic light scattering3 × mol/L Comparison of NanoDLSay with other methods for DNA detection Analytical Performance

11 Advantages Extremely simple Ultra-high sensitivity Excellent reproducibility Homogeneous solution assay Extremely easy to learn and use Data is easy to collect and interpret Require small volume of sample (<5 µL) No special sample preparation is required Applicable to a wide range of sample matrices Ultra-low cost of consumables to conduct the assay Results are obtained in minutes instead of hours or days Reveals new molecular information that cannot be or has not been detected by any other existing techniques NanoDLSay

12 NanoDLSay : The most comprehensive and powerful tool available for protein detection and analysis One new technology, discover a whole new world

13 X Individual protein monomer Protein complex Protein aggregates A B Limitations of traditional ELISA Comparisons Suitable for detecting individual proteins Not suitable when protein complexes are present antibody

14 Average particle size increase (nm) Incubation time (min) = 2D of analyte 0 min 30 min NanoDLSay Kinetic binding study: monitor the particle size change continuously during the assay Determine the size of the target analyte at a saturated binding level Determine if a target protein is a monomer, complex, or aggregates Label-free detection: no need to label the target proteins Detection of protein complexes and aggregates from real biological samples

15 NanoDLSay A two-step assay for protein complex detection and binding partner analysis Average particle size increase (nm) Incubation time (min) Step 2: Antibody screening Step 1: Catch the target Particle size change upon antibody addition c ~ 2D Binding partners Not binding partners 0 min 60 min

16 Co-Immunoprecipitation NanoDLSay: a two-step process Completed in ~30 min Co-Immunoprecipitation (Co-IP): Multiple steps (approximately steps) Takes hours to days to complete

17 Centrifuge Non-specific interactions Centrifuge in Co-IP process increases non- specific interactions Non-specific proteins are caught by the particles due to increased particle concentration towards the end of centrifuge This problem is not present in NanoDLSay NanoDLSay is more specific than Co-IP Co-IP and NanoDLSay : Which one is more specific?

18 Fluorescence techniques require the labeling of target proteins Size exclusion chromatography (SEC) and analytical ultracentrifuge (AU) are only suitable for pure protein solution study SEC may underestimate the aggregation level due to solvent elution; while AU may overestimate the aggregation level due to centrifuge-induced enrichment Label free Detects protein oligomers and aggregates directly from real biological samples Does not change the aggregation status of the sample during the assay High to ultra-high sensitivity NanoDLSay for label-free protein oligomer and aggregate detection from real biological samples Label-free detection NanoDLSay Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Other techniques

19 NDS1200: A new dynamic light scattering instrument designed for NanoDLSay Product & Services Automatic measurement of 12 samples Fast analysis time: 10-20s per sample 40 µL assay solution is used for the measurement Low-cost, disposable min-glass tubes with caps are used as sample containers. No cross-contamination between samples High throughput capability: samples/hour The hardware is maintenance-free No special housing environment is required

20 Product & Services NanoDLSay software: A software designed for convenient, flexible and high throughput analysis

21 Notes Patent application pending on NanoDLSay technology and NDS1200 system: PCT/US09/ and PCT/US11/21002 Nano Discovery Inc. has the exclusive license in the world to practice and commercialize NanoDLSay technology Please Contact Us to Request a Quote: NDS1200 Dynamic light scattering instrument for conducting NanoDLSay Assay kit including disposable sample cells and other consumables Research Parkway Suite 300 Orlando, FL Phone: NDS-Kit100 Order Information Or visit online: www. nanodiscoveryinc.com


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