Fabrication of a microfluidic enzyme reactor utilizing magnetic beads
2009
Liu, Xiaojun | Lo, Roger C. | Gomez, Frank A.
An enzyme-catalyzed microfluidic assay using magnetic micro-beads is described. Here, diaphorase (DI) (E.C. 1.6.99) is covalently attached to the magnetic micro-beads (2.7 μm) and integrated into a short section of a microchip fabricated from PDMS. DI converts non-fluorescent resazurin to fluorescent resorufin in the presence of nicotinamide adenine dinucleotide phosphate (NADH). In this work, an embedded magnet holds the micro-beads in place within the microchannel while a solution of resazurin and NADH in buffer is flowed through the beads. Incorporation of the micro-beads into the microchannel requires only a few minutes and offers well-defined spatial resolution and reproducibility. At a flow rate of 41.2 μL/h, a stable state for the enzyme reaction in the microfluidic format was achieved within 50 s. The maximum conversion of the reaction was obtained at a concentration of 1.25 mM NADH. The reaction yield is affected by ZnCl₂ and at concentrations in excess of 90.0 mM, the activity of DI was almost double without ZnCl₂. At 5.2 mM potassium chloride, the activity of DI reached its maximum value. Overall, the conversion of resazurin in microfluidic format was more than twice than that in a batch assay.
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