Ultrasensitive Digital Detection of Nanoparticles: Viral Diagnostics and Multiplexed Protein and Nucleic Acid Assays
Event details
| Date | 18.06.2014 |
| Hour | 10:15 |
| Speaker | Prof. M. Selim Ünlü, Boston University, Biomedical Engineering Department (Boston, MA, USA) |
| Location | |
| Category | Conferences - Seminars |
JOINT BIO- and ELECTRICAL ENGINEERING SEMINAR
Abstract:
Direct monitoring of primary molecular binding interactions without the need for secondary reactants would markedly simplify and expand applications of high-throughput label-free detection methods. We have developed the Interferometric Reflectance Imaging Sensor (IRIS) for label-free, high throughput, high sensitivity and dynamic detection of molecular binding on a solid surface. IRIS has demonstrated protein-protein binding and DNA-protein binding in real time, label-free, and in a high-throughput format with high sensitivity (~10 pg/mm2) and reproducibility [1,2] as well as label-free measurements of DNA hybridization kinetics [3] and viral detection [4].
Synthetic nanoparticles have made significant impact across a broad range of technological applications including optical nanoantennas, ultra-sensitive imaging and sensing, and diagnostics and therapeutics. Natural nanoparticles such as viruses and pollutants are involved in global health problems. High-throughput characterization of nanoparticles in terms of their size and shape is crucial for practical applications of synthetic nanoparticles and highly sensitive pathogen identification. Recently, we have demonstrated IRIS with the ability to detect single nanoscale particles [5,6].
In single-particle modality of IRIS (SP-IRIS), the interference of light reflected from the sensor surface is modified by the presence of particles producing a distinct signal that reveals the size of the particle. In our approach the dielectric layered structure acts as an optical antenna optimizing the elastic scattering characteristics of nanoparticles for sensitive detection and analysis. We have successfully detected 35 nm and 50 nm radius particles and H1N1 viruses with accurate size discrimination [5]. We have demonstrated identification of virus particles in complex samples for replication-competent wild-type vesicular stomatitis virus (VSV), defective VSV, and Ebola- and Marburg-pseudotyped VSV. Size discrimination of the imaged nanoparticles (virions) allows differentiation between modified viruses having different genome lengths and facilitates a reduction in the counting of non-specifically bound particles to achieve a limit-of-detection (LOD) of 5x103 pfu/mL for the Ebola and Marburg VSV pseudotypes. We have demonstrated the simultaneous detection of multiple viruses in serum or whole blood as well as in samples contaminated with high levels of bacteria [7]. Single nanoparticle detection with IRIS has shown promising results for protein [8] and DNA arrays with attomolar detection sensitivity.
References:
1. Ozkumur E., J. W. Needham, D. A. Bergstein, R. Gonzalez, M. Cabodi, J. M. Gershoni, B. B. Goldberg, and M. S. Ünlü. "Label-free and Dynamic Detection of Biomolecular Interactions for High-throughput Microarray Applications." Proceedings of the National Academy of Sciences, Vol. 105, No. 23, Pages: 7988-7992, (2008)
2. Özkumur E., A. Yalç?n, M. Cretich, C. A. Lopez, D. A. Bergstein, B. B. Goldberg, M. Chiari, and M. S. Ünlü. "Quantification of DNA and Protein Adsorption by Optical Phase Shift." Biosensors and Bioelectronics, Vol. 25, No. 1, 167-172, (2009)
3. E. Ozkumur, S. Ahn, A. Yalcin, C. Lopez, E. Cevik, R. Irani, C. DeLisi, M. Chiari, and M. S. Ünlü, "Label-free microarray imaging for direct detection of DNA hybridization and single-nucleotide mismatches," Biosensors and Bioelectronics, Vol. 25, No. 7, 15, pp. 1789-1795, (2010)
4. C. Lopez, G. G. Daaboul, R. S. Vedula, E. Ozkumur, D. A. Bergstein, T. W. Geisbert, H. Fawcett, B. B. Goldberg, J. H. Connor, and M. S. Ünlü, "Label-free multiplexed virus detection using spectral reflectance imaging," Biosensors and Bioelectronics, doi:10.1016/j.bios.2011.01.019, (2011)
5. G. G. Daaboul, A. Yurt, X. Zhang, G. M. Hwang, B. B. Goldberg, and M. S. Ünlü, "High-Throughput Detection and Sizing of Individual Low-Index Nanoparticles and Viruses for Pathogen Identification," Nano Letters, Vol. 10, No. 11, pp. 4727-4731 (2010)
6. A. Yurt, G. G. Daaboul, J. H. Connor, B. B. Goldberg, and M. S. Ünlü, "Single nanoparticle detectors for biological applications," Nanoscale, Vol. 4, No. 3, 2012, pp. 715 – 726
7. GG Daaboul, CA Lopez, J Chinnala, B Goldberg, JH Connor, and MS Ünlü, “Digital Sensing and Sizing of Vesicular Stomatitis Virus Pseudotypes in Complex Media: A Model for Ebola and Marburg Detection,” ACS Nano, 2014
8. M. R. Monroe, G. G. Daaboul, A. Tuysuzoglu, C. A. Lopez, F. F. Little, and M. S. Ünlü, "Single Nanoparticle Detection for Multiplexed Protein Diagnostics with Attomolar Sensitivity in Serum and Unprocessed Whole Blood," Analytical Chemistry, Vol. 85, No. 7, March/April 2013, pp. 3698-3706
Bio:
Currently at Boston University, Biomedical Engineering Department (Boston, MA, USA):
Professor, Biomedical Engineering
Professor, Electrical and Computer Engineering
Associate Director, Center for Nanoscience and Nanobiotechnology
Education:
M.S. & Ph.D., University of Illinois, Urbana-Champaign
B.S., Middle East Technical University, Ankara, Turkey
Abstract:
Direct monitoring of primary molecular binding interactions without the need for secondary reactants would markedly simplify and expand applications of high-throughput label-free detection methods. We have developed the Interferometric Reflectance Imaging Sensor (IRIS) for label-free, high throughput, high sensitivity and dynamic detection of molecular binding on a solid surface. IRIS has demonstrated protein-protein binding and DNA-protein binding in real time, label-free, and in a high-throughput format with high sensitivity (~10 pg/mm2) and reproducibility [1,2] as well as label-free measurements of DNA hybridization kinetics [3] and viral detection [4].
Synthetic nanoparticles have made significant impact across a broad range of technological applications including optical nanoantennas, ultra-sensitive imaging and sensing, and diagnostics and therapeutics. Natural nanoparticles such as viruses and pollutants are involved in global health problems. High-throughput characterization of nanoparticles in terms of their size and shape is crucial for practical applications of synthetic nanoparticles and highly sensitive pathogen identification. Recently, we have demonstrated IRIS with the ability to detect single nanoscale particles [5,6].
In single-particle modality of IRIS (SP-IRIS), the interference of light reflected from the sensor surface is modified by the presence of particles producing a distinct signal that reveals the size of the particle. In our approach the dielectric layered structure acts as an optical antenna optimizing the elastic scattering characteristics of nanoparticles for sensitive detection and analysis. We have successfully detected 35 nm and 50 nm radius particles and H1N1 viruses with accurate size discrimination [5]. We have demonstrated identification of virus particles in complex samples for replication-competent wild-type vesicular stomatitis virus (VSV), defective VSV, and Ebola- and Marburg-pseudotyped VSV. Size discrimination of the imaged nanoparticles (virions) allows differentiation between modified viruses having different genome lengths and facilitates a reduction in the counting of non-specifically bound particles to achieve a limit-of-detection (LOD) of 5x103 pfu/mL for the Ebola and Marburg VSV pseudotypes. We have demonstrated the simultaneous detection of multiple viruses in serum or whole blood as well as in samples contaminated with high levels of bacteria [7]. Single nanoparticle detection with IRIS has shown promising results for protein [8] and DNA arrays with attomolar detection sensitivity.
References:
1. Ozkumur E., J. W. Needham, D. A. Bergstein, R. Gonzalez, M. Cabodi, J. M. Gershoni, B. B. Goldberg, and M. S. Ünlü. "Label-free and Dynamic Detection of Biomolecular Interactions for High-throughput Microarray Applications." Proceedings of the National Academy of Sciences, Vol. 105, No. 23, Pages: 7988-7992, (2008)
2. Özkumur E., A. Yalç?n, M. Cretich, C. A. Lopez, D. A. Bergstein, B. B. Goldberg, M. Chiari, and M. S. Ünlü. "Quantification of DNA and Protein Adsorption by Optical Phase Shift." Biosensors and Bioelectronics, Vol. 25, No. 1, 167-172, (2009)
3. E. Ozkumur, S. Ahn, A. Yalcin, C. Lopez, E. Cevik, R. Irani, C. DeLisi, M. Chiari, and M. S. Ünlü, "Label-free microarray imaging for direct detection of DNA hybridization and single-nucleotide mismatches," Biosensors and Bioelectronics, Vol. 25, No. 7, 15, pp. 1789-1795, (2010)
4. C. Lopez, G. G. Daaboul, R. S. Vedula, E. Ozkumur, D. A. Bergstein, T. W. Geisbert, H. Fawcett, B. B. Goldberg, J. H. Connor, and M. S. Ünlü, "Label-free multiplexed virus detection using spectral reflectance imaging," Biosensors and Bioelectronics, doi:10.1016/j.bios.2011.01.019, (2011)
5. G. G. Daaboul, A. Yurt, X. Zhang, G. M. Hwang, B. B. Goldberg, and M. S. Ünlü, "High-Throughput Detection and Sizing of Individual Low-Index Nanoparticles and Viruses for Pathogen Identification," Nano Letters, Vol. 10, No. 11, pp. 4727-4731 (2010)
6. A. Yurt, G. G. Daaboul, J. H. Connor, B. B. Goldberg, and M. S. Ünlü, "Single nanoparticle detectors for biological applications," Nanoscale, Vol. 4, No. 3, 2012, pp. 715 – 726
7. GG Daaboul, CA Lopez, J Chinnala, B Goldberg, JH Connor, and MS Ünlü, “Digital Sensing and Sizing of Vesicular Stomatitis Virus Pseudotypes in Complex Media: A Model for Ebola and Marburg Detection,” ACS Nano, 2014
8. M. R. Monroe, G. G. Daaboul, A. Tuysuzoglu, C. A. Lopez, F. F. Little, and M. S. Ünlü, "Single Nanoparticle Detection for Multiplexed Protein Diagnostics with Attomolar Sensitivity in Serum and Unprocessed Whole Blood," Analytical Chemistry, Vol. 85, No. 7, March/April 2013, pp. 3698-3706
Bio:
Currently at Boston University, Biomedical Engineering Department (Boston, MA, USA):
Professor, Biomedical Engineering
Professor, Electrical and Computer Engineering
Associate Director, Center for Nanoscience and Nanobiotechnology
Education:
M.S. & Ph.D., University of Illinois, Urbana-Champaign
B.S., Middle East Technical University, Ankara, Turkey
Practical information
- Informed public
- Free
Organizer
- Prof. Yusuf Leblebici (Inst. of Electrical Engineering, STI)