Particle Interaction Monitoring
Particle Interaction Monitoring “Real-time detection of particle-particle, particle-biomolecule, functionalization and aggregation interactions.” qNano detects, monitors and analyzes interactions between nanoparticles and particles and biomolecules. Such interactions may result in a measurable change in key parameters: blockade event frequency, duration and magnitude. Binding reactions can result in the generation of products that exhibit differing electrophoretic properties from the original constituent particles. This information can be used to: • Confirm the occurrence of aggregation, binding interactions or particle functionalization • Detect the presence of molecules that may otherwise lie below the detection limit of the instrument • Monitor binding interactions in real-time, particle-by-particle, by combining reactants in the upper fluid cell.
www.izon.com
Measure Particle Aggregation
Small Molecule Detection and Diagnostics
The aggregation state of particles is important for a number of applications. In vaccinology knowledge of aggregation helps shorten the development time associated with optimizing virus preparation conditions and identifying stable formulations.
Molecules that otherwise may be too small to be detected by SIOS technology presently may be detected indirectly by monitoring key blockade event parameters. This principle may be used as a diagnostic tool using specific functionalisation of nanoparticles to detect molecules of interest in a sample through changes in blockade event parameters caused by specific binding of the target molecules to the nanoparticle surface.
Example: Stability Assessment of Viral Vaccines Izon provides an accurate method for counting and determining the size distribution of viruses in samples of unknown concentration. Separating viral particles by their size enables estimation of the amount of aggregated viral particles and yields a more accurate assessment for vaccine development companies (See Virus Quantitation Appliction Note).
Example: Metallic Nanoparticles in Diagnostics Research
Citrate ion-coated AuNP (~50 nm)
135.7
135.7
126.0
126.0
0nM PNA
Size (nm)
114.5
Nanopore-based detection identified subtle particle size changes induced by peptide nucleic acid (PNA, 22-mer) probe and gold nanoparticles (AuNP) interaction at very low concentration (5 nM). qNano enabled quantification of AuNP dynamic aggregation, with significant resolution advantages for metal nanoparticle measurement in comparison to light-based techniques.
100.0
79.4 50.0
79.4 50.0
0.0 0.5
1.0
1.5
2.0
2.5
Duration (ms)
PNA
DNA-PNA hybrids
Size (nm)
0.0 0.5
135.7
135.7 126.0
10nM PNA
1.5
2.0
2.5
25nM PNA
114.5
100.0
1.0
Duration (ms)
126.0 114.5
100.0
79.4 50.0
79.4 50.0
0.0 0.5
1.0
1.5
2.0
Duration (ms)
ABOVE: PNA-DNA hybridization results in unaggregated AuNPs, cf. PNA alone.
5nM PNA
114.5
100.0
2.5
0.0 0.5
1.0
1.5
2.0
Duration (ms)
2.5
Low et al. Aust. J. Chem. 2011, 64, 1229–1234
Quantify Functionalization The modification of particles by functionalization may result in a change in the measured blockade event frequency, duration or magnitude. Measurement of this change can be used to confirm surface modifications.
Example: PEGlyation of Liposomes (BELOW) In the example below the addition of PEG to the liposome surface can be detected as an increase in particle diameter and blockade duration (translocation time through the nanopore). The latter indicates a decrease in surface charge and electrophoretic mobility that occurs when PEG is added to the liposome surface. 10
8 7 6 5 4 3 2 1 0
60
80
100
120
140
160
180
200
Particle Diameter (nm)
Monitor Particle Interactions in Real-Time
20
9
Liposomes Liposomes (PEG)
8 7 6 5 4 3 2 1 0
1
3
5
7
9
13
15
Blockade Baseline Duration (ms)
Example: AVIDIN BINDING CARBOXYLATED POLYSTYRENE.
Particle-particle interactions can be carried out in situ in the upper fluid cell and the interaction monitored in real-time through changes in blockade duration or blockade magnitude. Izon Control Suite Software allows analysis of reaction dynamics including the ability to analyze, graph and report mean measurements as they occur over time.
11
In the example to the right, the addition of 10 ng/µL of Avidin to the upper fluid cell results in binding interactions with Carboxylated Polystyrene that can be detected as a change in the mean signal duration (ms) over time.
18
Liposomes Liposomes (PEG)
16 14 12 10 8 6 4 2 0
80 90 100 110 120 130 140 150 160 170 180 190
Particle Diameter (nm)
0.7
MEAN SIGNAL DURATION (ms)
Liposomes Liposomes (PEG)
% Population (by count)
% Population (by count)
9
Blockade Baseline Duration (ms)
10
0.6
Polystyrene particles only
0.5 0.4 0.3 0.2
10 ng/µL of Avidin introduced to the upper fluid cell
0.1 0 0
5
10
15
20
ELAPSED TIME (mins)
Enquire at [email protected] and ask how we can improve the quality of your particle analysis research.
www.izon.com
HEAD OFFICE 175 Roydvale Avenue, Burnside, Christchurch 8053, New Zealand Phone +64-3-357-4270 Fax+64-3-357-4273
UK / EUROPE The Oxford Science Park, Magdalen Centre, Robert Robinson Avenue, Oxford OX4 4GA, United Kingdom Phone +44-1865-309-666
UNITED STATES 85 Bolton Street, Cambridge MA 02140, United States Phone +1-617-945-5936 Fax +1-857-259-6623
www.izon.com
Measure Particle Aggregation
Small Molecule Detection and Diagnostics
The aggregation state of particles is important for a number of applications. In vaccinology knowledge of aggregation helps shorten the development time associated with optimizing virus preparation conditions and identifying stable formulations.
Molecules that otherwise may be too small to be detected by SIOS technology presently may be detected indirectly by monitoring key blockade event parameters. This principle may be used as a diagnostic tool using specific functionalisation of nanoparticles to detect molecules of interest in a sample through changes in blockade event parameters caused by specific binding of the target molecules to the nanoparticle surface.
Example: Stability Assessment of Viral Vaccines Izon provides an accurate method for counting and determining the size distribution of viruses in samples of unknown concentration. Separating viral particles by their size enables estimation of the amount of aggregated viral particles and yields a more accurate assessment for vaccine development companies (See Virus Quantitation Appliction Note).
Example: Metallic Nanoparticles in Diagnostics Research
Citrate ion-coated AuNP (~50 nm)
135.7
135.7
126.0
126.0
0nM PNA
Size (nm)
114.5
Nanopore-based detection identified subtle particle size changes induced by peptide nucleic acid (PNA, 22-mer) probe and gold nanoparticles (AuNP) interaction at very low concentration (5 nM). qNano enabled quantification of AuNP dynamic aggregation, with significant resolution advantages for metal nanoparticle measurement in comparison to light-based techniques.
100.0
79.4 50.0
79.4 50.0
0.0 0.5
1.0
1.5
2.0
2.5
Duration (ms)
PNA
DNA-PNA hybrids
Size (nm)
0.0 0.5
135.7
135.7 126.0
10nM PNA
1.5
2.0
2.5
25nM PNA
114.5
100.0
1.0
Duration (ms)
126.0 114.5
100.0
79.4 50.0
79.4 50.0
0.0 0.5
1.0
1.5
2.0
Duration (ms)
ABOVE: PNA-DNA hybridization results in unaggregated AuNPs, cf. PNA alone.
5nM PNA
114.5
100.0
2.5
0.0 0.5
1.0
1.5
2.0
Duration (ms)
2.5
Low et al. Aust. J. Chem. 2011, 64, 1229–1234
Quantify Functionalization The modification of particles by functionalization may result in a change in the measured blockade event frequency, duration or magnitude. Measurement of this change can be used to confirm surface modifications.
Example: PEGlyation of Liposomes (BELOW) In the example below the addition of PEG to the liposome surface can be detected as an increase in particle diameter and blockade duration (translocation time through the nanopore). The latter indicates a decrease in surface charge and electrophoretic mobility that occurs when PEG is added to the liposome surface. 10
8 7 6 5 4 3 2 1 0
60
80
100
120
140
160
180
200
Particle Diameter (nm)
Monitor Particle Interactions in Real-Time
20
9
Liposomes Liposomes (PEG)
8 7 6 5 4 3 2 1 0
1
3
5
7
9
13
15
Blockade Baseline Duration (ms)
Example: AVIDIN BINDING CARBOXYLATED POLYSTYRENE.
Particle-particle interactions can be carried out in situ in the upper fluid cell and the interaction monitored in real-time through changes in blockade duration or blockade magnitude. Izon Control Suite Software allows analysis of reaction dynamics including the ability to analyze, graph and report mean measurements as they occur over time.
11
In the example to the right, the addition of 10 ng/µL of Avidin to the upper fluid cell results in binding interactions with Carboxylated Polystyrene that can be detected as a change in the mean signal duration (ms) over time.
18
Liposomes Liposomes (PEG)
16 14 12 10 8 6 4 2 0
80 90 100 110 120 130 140 150 160 170 180 190
Particle Diameter (nm)
0.7
MEAN SIGNAL DURATION (ms)
Liposomes Liposomes (PEG)
% Population (by count)
% Population (by count)
9
Blockade Baseline Duration (ms)
10
0.6
Polystyrene particles only
0.5 0.4 0.3 0.2
10 ng/µL of Avidin introduced to the upper fluid cell
0.1 0 0
5
10
15
20
ELAPSED TIME (mins)
Enquire at [email protected] and ask how we can improve the quality of your particle analysis research.
www.izon.com
HEAD OFFICE 175 Roydvale Avenue, Burnside, Christchurch 8053, New Zealand Phone +64-3-357-4270 Fax+64-3-357-4273
UK / EUROPE The Oxford Science Park, Magdalen Centre, Robert Robinson Avenue, Oxford OX4 4GA, United Kingdom Phone +44-1865-309-666
UNITED STATES 85 Bolton Street, Cambridge MA 02140, United States Phone +1-617-945-5936 Fax +1-857-259-6623
