BioMEMS (Biological-MicroElectroMechanical Systems) are seen as the next generation platform for performing many different biologically-based assays in areas such as drug discovery, due to their potential for providing improved performance and low assay cost. However, in reports to date of BioMEMS used in high throughput screening (HTS), the potential improvements in throughput and costs have yet to be realized. This research focuses on developing new highly integrated polymer BioMEMS platforms to achieve advantages in the screening of libraries of drug candidates using parallelized infrared fluorescence assays. As a demonstration of the efficacy of the technology, inhibitors of L1 endonuclease (L1-EN) will be screened. L1-EN induces double-strand breaks via a TTTT'AA consensus sequence and can contribute to genetic damage responsible for cellular aging, cancer formation or progression. Therefore, discovery of inhibitors of L1-EN, of which there is currently none, could play an important role in minimizing genetic instability during cancer therapies or certain aspects of aging. The platforms to be developed have several enabling technologies that will allow the dissemination of HTS capabilities into the broader drug-discovery community by significantly reducing equipment demands and minimizing consumable costs. The system will consist of a large number of fluidic processors (96-192) configured on a 6"""""""" polymer wafer produced using micro- replication. Each fluidic processor will consist of (1) interconnected chip(s) to feed compounds directly from titer plates to the main processor wafer; (2) micromixer that speeds reagent mixing; (3) high surface area bioreactor in which the target is immobilized; and (4) grouped fluidic flow-cells for identifying potential leads using multi-channel fluorescence detection. A substrate of L1-EN will be dual-labeled with new, near-IR fluorescent phthalocyanine dyes, which provide high sensitivity and low background. Parallel fluorescence readout from the fluidic flow cells will be carried out by imaging a group of tightly spaced channels onto a CCD camera. Simulation and modeling will guide the selection of an appropriate readout format and optical configuration. In its finished format, the system will be able to screen -190,000 drug candidates in 24 hr. Fluid handling will be accomplished via microfluidics with minimal robotic intervention, as only a single sample transfer step is required to load the system with drug candidates. ? ? ?

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
1R01EB006639-01A1
Application #
7319369
Study Section
Instrumentation and Systems Development Study Section (ISD)
Program Officer
Korte, Brenda
Project Start
2007-08-01
Project End
2011-04-30
Budget Start
2007-08-01
Budget End
2008-04-30
Support Year
1
Fiscal Year
2007
Total Cost
$660,381
Indirect Cost
Name
Louisiana State University A&M Col Baton Rouge
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
075050765
City
Baton Rouge
State
LA
Country
United States
Zip Code
70803
Uba, Franklin I; Hu, Bo; Weerakoon-Ratnayake, Kumuditha et al. (2015) High process yield rates of thermoplastic nanofluidic devices using a hybrid thermal assembly technique. Lab Chip 15:1038-49
Canfield, Brian K; King, Jason K; Robinson, William N et al. (2014) Rapid, single-molecule assays in nano/micro-fluidic chips with arrays of closely spaced parallel channels fabricated by femtosecond laser machining. Sensors (Basel) 14:15400-14
Subramanian, Balamurugan; Kim, Namwon; Lee, Wonbae et al. (2011) Surface modification of droplet polymeric microfluidic devices for the stable and continuous generation of aqueous droplets. Langmuir 27:7949-57
Nesterova, Irina V; Bennett, Cecily A; Erdem, S Sibel et al. (2011) Near-IR single fluorophore quenching system based on phthalocyanine (Pc) aggregation and its application for monitoring inhibitor/activator action on a therapeutic target: L1-EN. Analyst 136:1103-5
Okagbare, Paul I; Soper, Steven Allan (2010) Polymer-based dense fluidic networks for high throughput screening with ultrasensitive fluorescence detection. Electrophoresis 31:3074-82
Lee, Wonbae; Obubuafo, Anne; Lee, Yong-Ill et al. (2010) Single-pair fluorescence resonance energy transfer (spFRET) for the high sensitivity analysis of low-abundance proteins using aptamers as molecular recognition elements. J Fluoresc 20:203-13
Robinson, William Neil; Davis, Lloyd M (2010) Simulation of single-molecule trapping in a nanochannel. J Biomed Opt 15:045006
Chantiwas, Rattikan; Hupert, Mateusz L; Pullagurla, Swathi R et al. (2010) Simple replication methods for producing nanoslits in thermoplastics and the transport dynamics of double-stranded DNA through these slits. Lab Chip 10:3255-64
Okagbare, Paul I; Emory, Jason M; Datta, Proyag et al. (2010) Fabrication of a cyclic olefin copolymer planar waveguide embedded in a multi-channel poly(methyl methacrylate) fluidic chip for evanescence excitation. Lab Chip 10:66-73
Lee, Wonbae; Lee, Yong-Ill; Lee, Jeonghoon et al. (2010) Cross-talk-free dual-color fluorescence cross-correlation spectroscopy for the study of enzyme activity. Anal Chem 82:1401-10

Showing the most recent 10 out of 16 publications