Objective: The goal of proposed research is to fabricate and use highly piezoelectric microcantilever arrays for ultrasensitive, in-situ, rapid, simultaneous multiple analyte quantification in small sample volumes using electrical means with unprecedented sensitivity (10 -15 g). The proposed piezoelectric microcantilevers with antibodies specific to the target antigen immobilized at the cantilever tip will measure the presence of proteins or pathogens with femtogram (smaller than the mass of a single cell) sensitivity. This represents the ability to detect a single cell or about 1000 molecules in a small volume. We will demonstrate the application with (1) transient protein expression of a recombinant protein, alkaline phosphatase (SEAP) in Chinese Hamster Ovary cells (CHO) during growth in a bioreactor, and (2) early detection of streptococcus at ultra low concentrations , 100 - 10,000 number/ml. Approaches: The device consists of a highly piezoelectric lead magnesium niobate-lead titanate solid solution (PMN-PT) cantilever smaller than 50 um in length coupled to antibody immobilized at the cantilever tip. Binding of target antigens is detected by monitoring the resonance frequency shift. Because of the small sizes, the cantilevers will be capable of detecting a single cell or some 1000 molecules in a small volume. The resonance frequency shift transient will be used to quantify the amount of antigens present in a small volume. In the model CHO bioreactor, the SEAP expression quantification is used as an example to illustrate the proposed methodology. For pathogen detection, microcantilevers developed under this program have the potential to detect streptococcus at a concentration of 100 to 10,000 bacterial/ml, significantly enhancing detection capability. Experiments will be designed to explore lower level of detection in the range of 100 to 10,000 cells/ml. Because the proposed piezoelectric cantilever sensors use electrical signal for actuation and detection, the sensor and all necessary electronics can be organized in a compact form and easily usable in such broad ranging applications such as early disease detection and genomics-inspired proteomics. Results: It is anticipated that as a result of the proposed study, ultra sensitive, rapid, specific, multiple analyte quantification in ultra low concentrations and small sample volumes will be achieved using arrays of highly piezoelectric PMN-PT micro cantilevers of less than 50 um in length with better than10-15 g/Hz sensitivity coupled with antibodies specific to the target antigen immobilized at the cantilever tip with simple electrical means, which has wide ranging applications such early disease detection and genomics-inspired proteomics.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
5R01EB000720-03
Application #
6787152
Study Section
Special Emphasis Panel (ZRG1-SSS-F (02))
Program Officer
Korte, Brenda
Project Start
2002-09-30
Project End
2006-08-31
Budget Start
2004-09-01
Budget End
2006-08-31
Support Year
3
Fiscal Year
2004
Total Cost
$361,427
Indirect Cost
Name
Drexel University
Department
Engineering (All Types)
Type
Schools of Engineering
DUNS #
002604817
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Capobianco, Joseph A; Shih, Wan Y; Adams, Gregory P et al. (2011) Label-free Growth Receptor-2 Detection and Dissociation Constant Assessment in Diluted Human Serum Using a Longitudinal Extension Mode of a Piezoelectric Microcantilever Sensor. Sens Actuators B Chem 160:349-356
Li, Hui; Li, Mengyan; Shih, Wan Y et al. (2011) Cytotoxicity tests of water soluble ZnS and CdS quantum dots. J Nanosci Nanotechnol 11:3543-51
Yegingil, Hakki; Shih, Wan Y; Shih, Wei-Heng (2010) Probing model tumor interfacial properties using piezoelectric cantilevers. Rev Sci Instrum 81:095104
Li, Hui; Shih, Wan Y; Shih, Wei-Heng (2010) Highly Photoluminescent and Stable Aqueous ZnS Quantum Dots. Ind Eng Chem Res 49:579-582
Capobianco, Joseph A; Shih, Wei-Heng; Leu, Jiann-Horng et al. (2010) Label free detection of white spot syndrome virus using lead magnesium niobate-lead titanate piezoelectric microcantilever sensors. Biosens Bioelectron 26:964-9
McGovern, John-Paul; Shih, Wei-Heng; Rest, Richard F et al. (2009) Array lead zirconate titanate/glass piezoelectric microcantilevers for real-time detection of Bacillus anthracis with 10 spores/ml sensitivity and 1/1000 selectivity in bacterial mixtures. Rev Sci Instrum 80:125104
Zhu, Qing; Shih, Wan Y; Shih, Wei-Heng (2009) Enhanced Detection Resonance Frequency Shift of a Piezoelectric Microcantilever Sensor by a DC Bias Electric Field in Humidity Detection. Sens Actuators B Chem 138:1-4
Capobianco, Joseph A; Shih, Wan Y; Yuan, Qing-An et al. (2008) Label-free, all-electrical, in situ human epidermal growth receptor 2 detection. Rev Sci Instrum 79:076101
McGovern, John-Paul; Shih, Wan Y; Rest, Richard et al. (2008) Label-free flow-enhanced specific detection of Bacillus anthracis using a piezoelectric microcantilever sensor. Analyst 133:649-54
Zhu, Qing; Shih, Wan Y; Shih, Wei-Heng (2008) Mechanism of flexural resonance frequency shift of a piezoelectric microcantilever sensor during humidity detection. Appl Phys Lett 92:183505-1835053

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