19-09-2013 дата публикации
Номер: US20130244257A1
An immunocapture-based in vitro kinase assay on an integrated polydimethylsiloxane (PDMS) microfluidics platform that can reproducibly measure kinase activity from as few as 3,000 cells is described. For this platform, the standard radiometric P-ATP labeled phosphate transfer assay was adopted. Implementation on a microfluidic device required the development of methods for repeated trapping and mixing of solid-phase affinity micro beads. A solid state beta-particle camera imbedded directly below the microfluidic device was used to provide real-time quantitative detection of the signal from this and other microfluidic radio bioassays. The integrated device can measure ABL protein kinase activity from BCR-ABL positive leukemia patient samples, and can measure the small molecule phosphorylation such as phosphorylation of the deoxycytidine analog F-FAC by deoxycytidine kinase (dCK) isolated from cell lysates. 1. An apparatus for performing a microfluidic radioactivity based assay , comprising:a microfluidic chip with a microchannel system and at least one reaction element adapted to capture radioactively labeled molecules; anda solid state radiation detector coupled to the microfluidic chip;wherein radiation emitted from said microfluidic chip is detected.2. An apparatus as recited in claim 1 , wherein said reaction element comprises:a plurality of reaction columns retaining affinity beads fluidly connected to the microchannel system of the microchip.3. An apparatus as recited in claim 1 , wherein said reaction element comprises:a plurality of reaction columns retaining surface functionalized beads fluidly connected to the microchannel system of the microchip.4. An apparatus as recited in claim 3 , wherein said surface functionalized beads are functionalized with a coating of a plurality of enzyme specific antibodies claim 3 , wherein enzymes disposed in a liquid flow through the reaction column bind to the antibodies on the surface of the beads.5. An apparatus as ...
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