COMBINED VACUUM ULTRAFILTRATION--SEPARATION OF ENDOTOXIN

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Endotoxin, a lipopolysaccharide product of gram negative bacteria, has profound toxicities in vivo (e.g., cardiovascular collapse due to endotoxin shock) as well as in a variety of in vitro biological systems (e.g., tissue culture). The current technology available to remove endotoxin form biological is a particle based affinity chromatography procedure utilizing Sepharose-Polymyxin. As is typical for particle based separations this process is slow ad inconvenient due to the changing characteristics of affinity columns with age (e.g., packing). In this proposal we outline studies designed to optimize surface chemistries for attachment of Polymyxin to non-particle based supports. These supports are rigid, have excellent flow rates and, thus, will have physical properties compatible with high volume applications, such as detoxification of pharmaceuticals. Once the attachment chemistry is defined we will determine the optimum configuration of the Polymyxin support in combination with vacuum ultrafiltration to provide a single unit which will both sterilize and detoxify in a single step. If successful this research should lay the groundwork for development of a prototype device which, when refined, should find widespread uses in the Biotechnology and Pharmaceutical industries, to name a few. Additional positive features of such a device is that it will be inexpensive, convenient to use (perhaps even re-useable) and amenable to interfacing with other flow-thru processes.