Abstract
A nanoarray based-single molecule detection system was developed for detecting proteins with extremely high sensitivity. The nanoarray was able to effectively trap nanoparticles conjugated with biological sample into nanowells by integrating with an electrophoretic particle entrapment system (EPES). The nanoarray/EPES is superior to other biosensor using immunoassays in terms of saving the amounts of biological solution and enhancing kinetics of antibody binding due to reduced steric hindrance from the neighboring biological molecules. The nanoarray patterned onto a layer of PMMA and LOL on conductive and transparent indium tin oxide (ITO)-glass slide by using e-beam lithography. The suspension of 500 nm-fluorescent (green emission)-carboxylated polystyrene (PS) particles coated with protein-A followed by BDE 47 polyclonal antibody was added to the chip that was connected to the positive voltage. The droplet was covered by another ITO-coated-glass slide and connected to a ground terminal. After trapping the particles into the nanowells, the solution of different concentrations of anti-rabbit- IgG labeled with Alexa 532 was added for an immunoassay. A single molecule detection system could quantify the anti-rabbit IgG down to atto-mole level by counting photons emitted from the fluorescent dye bound to a single nanoparticle in a nanowell.
| Original language | English (US) |
|---|---|
| Title of host publication | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
| Volume | 7908 |
| DOIs | |
| State | Published - 2011 |
| Event | Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VIII - San Francisco, CA, United States Duration: Jan 24 2011 → Jan 27 2011 |
Other
| Other | Nanoscale Imaging, Sensing, and Actuation for Biomedical Applications VIII |
|---|---|
| Country | United States |
| City | San Francisco, CA |
| Period | 1/24/11 → 1/27/11 |
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Keywords
- ebeam lithography
- electrophoretic particle entrapment system (EPES)
- nanoarray
- photons
- single molecule detection system
ASJC Scopus subject areas
- Atomic and Molecular Physics, and Optics
- Electronic, Optical and Magnetic Materials
- Biomaterials
- Radiology Nuclear Medicine and imaging
Cite this
Highly sensitive immunoassay of protein molecules based on single nanoparticle fluorescence detection in a nanowell. / Han, Jin Hee; Kim, Hee Joo; Lakshmana, Sudheendra; Gee, Shirley J.; Hammock, Bruce D.; Kennedy, Ian M.
Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 7908 2011. 79080S.Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
}
TY - GEN
T1 - Highly sensitive immunoassay of protein molecules based on single nanoparticle fluorescence detection in a nanowell
AU - Han, Jin Hee
AU - Kim, Hee Joo
AU - Lakshmana, Sudheendra
AU - Gee, Shirley J.
AU - Hammock, Bruce D.
AU - Kennedy, Ian M.
PY - 2011
Y1 - 2011
N2 - A nanoarray based-single molecule detection system was developed for detecting proteins with extremely high sensitivity. The nanoarray was able to effectively trap nanoparticles conjugated with biological sample into nanowells by integrating with an electrophoretic particle entrapment system (EPES). The nanoarray/EPES is superior to other biosensor using immunoassays in terms of saving the amounts of biological solution and enhancing kinetics of antibody binding due to reduced steric hindrance from the neighboring biological molecules. The nanoarray patterned onto a layer of PMMA and LOL on conductive and transparent indium tin oxide (ITO)-glass slide by using e-beam lithography. The suspension of 500 nm-fluorescent (green emission)-carboxylated polystyrene (PS) particles coated with protein-A followed by BDE 47 polyclonal antibody was added to the chip that was connected to the positive voltage. The droplet was covered by another ITO-coated-glass slide and connected to a ground terminal. After trapping the particles into the nanowells, the solution of different concentrations of anti-rabbit- IgG labeled with Alexa 532 was added for an immunoassay. A single molecule detection system could quantify the anti-rabbit IgG down to atto-mole level by counting photons emitted from the fluorescent dye bound to a single nanoparticle in a nanowell.
AB - A nanoarray based-single molecule detection system was developed for detecting proteins with extremely high sensitivity. The nanoarray was able to effectively trap nanoparticles conjugated with biological sample into nanowells by integrating with an electrophoretic particle entrapment system (EPES). The nanoarray/EPES is superior to other biosensor using immunoassays in terms of saving the amounts of biological solution and enhancing kinetics of antibody binding due to reduced steric hindrance from the neighboring biological molecules. The nanoarray patterned onto a layer of PMMA and LOL on conductive and transparent indium tin oxide (ITO)-glass slide by using e-beam lithography. The suspension of 500 nm-fluorescent (green emission)-carboxylated polystyrene (PS) particles coated with protein-A followed by BDE 47 polyclonal antibody was added to the chip that was connected to the positive voltage. The droplet was covered by another ITO-coated-glass slide and connected to a ground terminal. After trapping the particles into the nanowells, the solution of different concentrations of anti-rabbit- IgG labeled with Alexa 532 was added for an immunoassay. A single molecule detection system could quantify the anti-rabbit IgG down to atto-mole level by counting photons emitted from the fluorescent dye bound to a single nanoparticle in a nanowell.
KW - ebeam lithography
KW - electrophoretic particle entrapment system (EPES)
KW - nanoarray
KW - photons
KW - single molecule detection system
UR - http://www.scopus.com/inward/record.url?scp=79953158612&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79953158612&partnerID=8YFLogxK
U2 - 10.1117/12.874259
DO - 10.1117/12.874259
M3 - Conference contribution
AN - SCOPUS:79953158612
SN - 9780819484451
VL - 7908
BT - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
ER -