Comparison of Tissue Contractile Responses of Rat Models of Pulmonary Artery Hypertension and Type I/II Diabetes

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Authors

Islam, Schinria R.
Bridges, Fredy
Laxa, Madeline
McKenna, Becky
Pugh, Shawn

Issue Date

2010

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Thesis

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en_US

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Research Projects

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Abstract

Protein purification processes have become highly sophisticated in recent decades. Even the simplest separations require various steps to ensure the highest possible purity and maximum profit. The goal of this paper is to propose a configuration of unit operations that will purify a monoclonal antibody (Lucentis) grown and extracted from e. coli for the purpose of treating the “wet” form of macular degeneration. The following units will be explored, in this determined order: homogenizer, centrifuge, column chromatography, tangential flow filtration. Specifications for these units will be determined using data from either literature research or experimentation. A sample of bacteria culture that has been processed through a homogenizer and centrifuge were obtained and used in our experimentations. A homogenizer is used to break apart the cell wall to release the proteins inside. A centrifuge is used to do a crude mass separation before more intrinsic steps to isolate the desired antibody are needed. Specifications for these two units such as reactor size and flow area will be theorized. Two types of column chromatography will be used to separate and purify the sample: cation exchange and mixed (hydrophobic interactionanion exchange) mode. The resin height, column height, and column diameter will be determined for these units. Chromatography is commonly used to isolate and purify proteins. A tangential flow filtration is then used to remove any excess debris from the sample as the last purification procedure. The culmination of these units proposes a purification process that can ideally isolate a desired antibody and produce a pure product, ready for human use.

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