title: Integrated bioassays in microfluidic devices: botulinum toxin assays.
abstract: A microfluidic assay was developed for screening botulinum neurotoxin serotype A (BoNT-A) by using a fluorescent resonance energy transfer (FRET) assay. Molded silicone microdevices with integral valves, pumps, and reagent reservoirs were designed and fabricated. Electrical and pneumatic control hardware were constructed, and software was written to automate the assay protocol and data acquisition. Detection was accomplished by fluorescence microscopy. The system was validated with a peptide inhibitor, running 2 parallel assays, as a feasibility demonstration. The small footprint of each bioreactor cell (0.5 cm2) and scalable fluidic architecture enabled many parallel assays on a single chip. The chip is programmable to run a dilution series in each lane, generating concentration-response data for multiple inhibitors. The assay results showed good agreement with the corresponding experiments done at a macroscale level. Although the system has been developed for BoNT-A screening, a wide variety of assays can be performed on the microfluidic chip with little or no modification.
authors.affiliation: Sarnoff Corporation, 201 Washington Road, Princeton, NJ 08543-5300, USA.
authors.forename: Shakuntala
authors.initials: S
authors.lastname: Mangru
authors.forename: Bryan L
authors.initials: BL
authors.lastname: Bentz
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Davis
authors.forename: Nitin
authors.initials: N
authors.lastname: Desai
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Stabile
authors.forename: James J
authors.initials: JJ
authors.lastname: Schmidt
authors.forename: Charles B
authors.initials: CB
authors.lastname: Millard
authors.forename: Sina
authors.initials: S
authors.lastname: Bavari
authors.forename: Krishna
authors.initials: K
authors.lastname: Kodukula
mesh_terms: D001681:Biological Assay / Q000295:instrumentation / Q000379:methods*; D019274:Botulinum Toxins, Type A / Q000032:analysis*; D005240:Feasibility Studies; D008832:Microchemistry / Q000295:instrumentation; D044085:Microfluidics / Q000295:instrumentation*; D008904:Miniaturization
chemical_list: D019274:Botulinum Toxins, Type A