Enzymatic and cellular assays on a microfabricated system

Brian Joseph Burke, Purdue University

Abstract

Microfabricated devices are being described to perform a variety of tasks including biochemical assays, process monitoring, off-site testing, and healthcare diagnostics. The work presented here focused on the use of these devices to perform enzymatic and cellular assays. In typical open channel, microfabricated devices, the primary method of mixing reagents is through the use of simple diffusion. While convenient, diffusion is shown to be an inefficient method to mix reagents. Several methods were proposed to overcome this diffusional mixing situation. One technique utilized here is known as EMMA (Electrophoretically Mediated Micro-Analysis) which performs enzymatic assays in open channels. In EMMA, the reagents are mixed using the difference in the electrophoretic mobility of each component. A microfabricated device also was described and tested that eliminated the need to use diffusionally based mixing systems. Using this device, stopped flow enzymatic assays were performed that were compared to an external T-mixing system. Not only did the results compare well from the microfabricated device to the external T-mixer, but also a diffusionally based mixing system could not be used to obtain quantitative results. A second type of assay was performed on chips using live Escherichia Coli cells. The cells were trapped in a micro-chromatography column on a chip using small particles called COMOSS (Collocated Monolith Support Structures). An enzyme produced in the cells was assayed and could be used for over 10 hours.

Degree

Ph.D.

Advisors

Regnier, Purdue University.

Subject Area

Analytical chemistry|Biochemistry

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