Transcript Document

One new technology,
discover a whole new world
NanoDLSay™
Nanoparticle-Enabled Dynamic Light Scattering Assay for
Biological and Chemical Detection and Analysis
www.nanodiscoveryinc.com
12565 Research Parkway Suite 300
Orlando, Florida
Email: [email protected]
Copyright Nano Discovery Inc. 2012
One new technology,
discover a whole new world
NanoDLSay™: a new analytical
technique that uses particle size change
for signal transduction
Gold nanoparticles (AuNPs) are used as
the optical probe in NanoDLSay™
When analytes are bound to the nanoparticles, this
will cause various changes to the particle size…
Citrate-protected
AuNP (10-100 nm)
Y
Y
Y
A small chemical
(< a few nm)
Liu X, Dai Q, Austin L, Coutts J, Knowles G, J, Chen H, H Q. A One-step
D  130-160 nm
D > 130-160 nm
D = 100 nm
D  120 nm
homogeneous
immunoassay
for
cancer
biomarker
detection
using
gold
A protein monomer
(~5-20 nm)
nanoparticle
probes coupled with dynamic light
J. Am.AuNP
Chem.
Soc.
immunoprobe
AuNP scattering.
immune probe
Unmodified AuNP
AuNP immunoprobe
bound with a large protein
bound with a small protein
2008; 130:2780-2782.
complex
monomer
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: 81382+
A protein complex
8139.
(> 20 nm)
2+
Immunoglobulin G
(IgG) (~7-10 nm)
2+
2+
D >> 100 nm
AuNPs bound with metal ion targets
through metal-chelating ligands
Viruses (~10s nm)
AuNPs bound with small chemical targets
through coordinative ligand interactions
NanoDLSay™ Procedure
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One-step homogenous solution assay
Required sample volume: 1-5 µL
Results obtain in a few minutes
APPLICATIONS
Proteins
DNAs
RNAs
Viruses
NanoDLSay™
Small
chemicals
Toxic
metal ions
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; 130:2780-2782.
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: 38-44.
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: 9425-9432.
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, 10-12.
Bogdanovic J, Colon J, Baker C, Huo Q. A label-free nanoparticle aggregation assay for protein
complex/aggregate detection and analysis. Anal. Biochem. 2010; 45:96-102.
Huo Q. Protein complexes/aggregates as potential cancer biomarkers revealed by a nanoparticle
aggregation assay. Colloids Surfaces B 2010; 78:259-265.
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, 11047-11049. 
Wang, X.; Ramström, O.; Yan, M. Dynamic light scattering as an efficient tool to study glyconanoparticlelectin interactions. Analyst 2011, 136, 4174-4178.
Publications
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: 8138-8139.
 ao D, Sheng Z, Han H. An ultrasensitive method for the detection of gene fragment from transgenics
G
using label-free gold nanoparticle probe and dynamic light scattering. Anal. Chim Acta 2011; 696:15. 
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, 11230-11236. 
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; 22:285501 (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; 136:3083-3090.
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; 3:59-61. 
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; 2:3455-3460. 
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; 48:9668-9671.
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; 3:668-673.
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; 47:4192-4194.
Zhang L, Yao Y, Shan J, Li H. Lead (II) ion detection in surface water with pM sensitivity using azacrown-ether-modified silver nanoparticles via dynamic light scattering. Nanotechnology 2011;
22:275504 (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, 582-586.
Xiong C, Ling L. Label-free, sensitive detection of Hg(II) with gold nanoparticles by using dynamic light
scattering technique. Talanta 2012, 89, 317-321.
Analytical Performance
Analytes
Sensitivity
Dynamic Range
Proteins
High pg/mL to low ng/mL range
2-3 orders of magnitude
DNAs
30 fM
(5 orders of magnitude more sensitive than SPR
and fluorescence techniques)
> 5 orders of magnitude
Viruses
< 100 TCID50/mL
(1-2 orders of magnitude more sensitive than
commercial diagnostic kits)
2-3 orders of magnitude
Toxic metal ions
Arsenics: 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 molecules
7 nM
(5 orders of magnitude more sensitive than the
colorimetric method)
> 4 orders of magnitude
Explosive chemicals
100 pM
2-3 orders of magnitude
Notes:
(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.
Analytical Performance
Comparison of NanoDLSay™ with other methods for DNA detection
Label
Method
Detection limit
AuNP
Colorimetric
1 × 10-8 mol/L
Au chip
Surface plasmon resonance
1 × 10-9 mol/L
Au/polyaniline
nantube
Electrochemical impedance
spectroscopy
3 × 10-13 mol/L
Quantum dots
Anodic stripping voltammetry
5 × 10-11 mol/L
ZnS and CdSe
quantum dots
Fluorescence
2 × 10-9 mol/L
NanoDLSay™
Dynamic light scattering
3 × 10-14 mol/L
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.
Advantages
NanoDLSay™
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
One new technology,
discover a whole new world
NanoDLSay™: The most comprehensive
and powerful tool available for protein
detection and analysis
Comparisons
Limitations of traditional ELISA
Individual protein
monomer
A
Protein complex
antibody
Protein aggregates
B
X
Suitable for detecting
individual proteins
Not suitable when protein
complexes are present
NanoDLSay™
3
Average particle size increase (nm)
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
2

1

 = 2D of analyte
0 min
Incubation time (min)
30 min
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
NanoDLSay™
Average particle size increase (nm)
A two-step assay for protein complex detection and
binding partner analysis
Step 2: Antibody
screening
Step 1: Catch the
target
Binding partners
Particle size change
upon antibody addition
c
 ~ 2D
0 min
Incubation time (min)
Not binding partners
60 min
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NanoDLSay™: a two-step process
Completed in ~30 min
Co-Immunoprecipitation
Co-Immunoprecipitation (Co-IP):
 Multiple steps (approximately 15-20 steps)
 Takes hours to days to complete
Non-specific interactions
Co-IP and NanoDLSay™:
Which one is more specific?

Centrifuge in Co-IP
process increases nonspecific 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
Label-free detection
NanoDLSay™ for label-free protein oligomer and aggregate
detection from real biological samples
Y
Y
Y
Y
Other techniques
NanoDLSay™
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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
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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
Product & Services
NDS1200: A new dynamic light scattering
instrument designed for NanoDLSay™
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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: 120-180
samples/hour
The hardware is maintenance-free
No special housing environment is
required
Product & Services
NanoDLSay™ software: A software designed for convenient,
flexible and high throughput analysis
Order Information
NDS1200
Dynamic light scattering instrument for conducting NanoDLSay™
NDS-Kit100
Assay kit including disposable sample cells and other consumables
Please Contact Us to Request a Quote:
12565 Research Parkway Suite 300
Orlando, FL 32826
Phone: 407-770-8954
Email: [email protected]
Or visit online:
www. nanodiscoveryinc.com
Notes
Patent application pending on NanoDLSay™ technology and NDS1200 system: PCT/US09/030087 and PCT/US11/21002
Nano Discovery Inc. has the exclusive license in the world to practice and commercialize NanoDLSay™ technology