The goal of the proposed research is the development of reliable and widely applicable methods for the multiplexed in vitro detection of protein biomarkers at extremely low (femtomolar) concentrations in microliter sample volumes. These novel ultrasensitive microarray biosensing measurements will be achieved through the combined use of surface enzyme chemistries, biofunctionalized nanoparticles, and unique surface plasmon-based bioaffinity adsorption detection techniques. Key components in these new methodologies are (i) the use of enzymatic reactions for the attachment, modification and amplification of biomolecules on both nanoparticle surfaces and microarray elements to increase the selectivity and sensitivity of the bioaffinity sensing process, (ii) the incorporation of chemically and enzymatically biofunctionalized nanoparticles into the microarray sensing measurements in order to enhance the capture of and signal generated by target biomolecules, and (iii) the implementation of novel optical detection methods such as surface plasmon resonance phase imaging and nanoparticle-enhanced optical diffraction that provide additional biosensing sensitivity in a multiplexed format. A combination of aptamers, antibodies and enzymatic attachment reactions will be used to capture and detect protein (e.g., VEGF, PSA, troponin) and microRNA biomarker targets that exist in biofluid samples at zeptomole levels. These new detection methodologies that can identify and quantitate multiple proteins at femtomolar concentrations and below are essential technologies for the discovery of new sets of protein biomarkers, and the ability to track these new biomarkers (e.g., cardiovascular and cancer biomarkers) at such low levels will facilitate the creation of better methods for early disease detection and strategies for post-treatment patient monitoring.

Public Health Relevance

Novel methodologies will be created and demonstrated for the simultaneous, multiplexed detection and identification of extremely small amounts (down to thousands of molecules) of biologically relevant proteins and nucleic acids, also known as biomarkers. These ultrasensitive biosensing technologies are essential for the discovery of new sets of protein biomarkers that are only present at minute concentration levels in biological fluid samples, and will facilitate the creation of new methods for earlier and more precise disease detection (e.g., cancers, heart disease), as well as the creation of better post-treatment patient monitoring strategies.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM059622-14
Application #
8517734
Study Section
Special Emphasis Panel (ZRG1-IMST-L (90))
Program Officer
Lewis, Catherine D
Project Start
1999-08-01
Project End
2015-07-31
Budget Start
2013-08-01
Budget End
2014-07-31
Support Year
14
Fiscal Year
2013
Total Cost
$294,752
Indirect Cost
$92,102
Name
University of California Irvine
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
046705849
City
Irvine
State
CA
Country
United States
Zip Code
92697
Fasoli, Jennifer B; Corn, Robert M (2015) Surface Enzyme Chemistries for Ultrasensitive Microarray Biosensing with SPR Imaging. Langmuir 31:9527-36
Cho, Kyunghee; Fasoli, Jennifer B; Yoshimatsu, Keiichi et al. (2015) Measuring melittin uptake into hydrogel nanoparticles with near-infrared single nanoparticle surface plasmon resonance microscopy. Anal Chem 87:4973-9
Sharma, Himanshu; Wood, Jennifer B; Lin, Sophia et al. (2014) Shrink-induced silica multiscale structures for enhanced fluorescence from DNA microarrays. Langmuir 30:10979-83
Loget, Gabriel; Corn, Robert M (2014) Silica nanowire arrays for diffraction-based bioaffinity sensing. Chemistry 20:10802-10
Cho, Kyunghee; Loget, Gabriel; Corn, Robert M (2014) Lithographically Patterned Nanoscale Electrodeposition of Plasmonic, Bimetallic, Semiconductor, Magnetic, and Polymer Nanoring Arrays. J Phys Chem C Nanomater Interfaces 118:28993-29000
Chen, Yulin; Corn, Robert M (2013) DNAzyme footprinting: detecting protein-aptamer complexation on surfaces by blocking DNAzyme cleavage activity. J Am Chem Soc 135:2072-5
Wood, Jennifer B; Szyndler, Megan W; Halpern, Aaron R et al. (2013) Fabrication of DNA microarrays on polydopamine-modified gold thin films for SPR imaging measurements. Langmuir 29:10868-73
Seefeld, Ting H; Halpern, Aaron R; Corn, Robert M (2012) On-chip synthesis of protein microarrays from DNA microarrays via coupled in vitro transcription and translation for surface plasmon resonance imaging biosensor applications. J Am Chem Soc 134:12358-61
Chen, Yulin; Nakamoto, Kohei; Niwa, Osamu et al. (2012) On-chip synthesis of RNA aptamer microarrays for multiplexed protein biosensing with SPR imaging measurements. Langmuir 28:8281-5
Seefeld, Ting Hu; Zhou, Wen-Juan; Corn, Robert M (2011) Rapid microarray detection of DNA and proteins in microliter volumes with surface plasmon resonance imaging measurements. Langmuir 27:6534-40

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