Date of Graduation

5-2016

Document Type

Thesis

Degree Name

Bachelor of Science in Biomedical Engineering

Degree Level

Undergraduate

Department

Biomedical Engineering

Advisor

Stenken, Julie

Reader

Kim, Michelle

Second Reader

Balachandran, Kartik

Abstract

Microdialysis sampling uses a semi-permeable membrane to allow solute collection by diffusion. When used in conjunction with other instruments, analytes in question can be quantified. Matrix Metalloproteinases (MMPs) are enzymes involved in numerous biological processes where they serve the role of degrading extracellular matrix. Microdialysis sampling, in coordination with further analysis methods, can be used in order to measure the activity of these enzymes in a region of the body instantaneously. The intention of this project is to determine ways of measuring in vitro activity of elastase from porcine pancreas using determined activity values and the collection of elastase products. This is a necessary step in the advancement of this technology towards in vivo research. Elastase from the porcine pancreas is a substitute for MMP in vivo. Experiments were performed using succinyl(Ala-p-nitroaniline, 4-nitroanaline, and elastase from porcine pancreas. The Nanodrop, a UV-Vis instrument, was used in order to measure the concentration of p-nitroaniline (p-NA) using its maximum absorbance at a wavelength of 380 nm. A microdialysis probe with a molecular cutoff of 20 kDa was used in order to collect p-NA from solution and transfer it into collection vials for analyzation of p-NA concentration. The concentration of p-NA increased over time until it reached a point where it would level off. This level-off value varied based upon the concentration of the suc-(Ala)3-pNA perfusate. Experiments showed a decrease in activity of elastase with decreasing concentrations or increasing storage time. In microdialysis sampling experiments, the concentration of suc-(Ala)3-pNA in the samples collected showed a decrease as the concentration of the pNA collected increased.

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