COMBINED VACUUM ULTRAFILTRATION--SEPARATION OF ENDOTOXIN

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 FROM BIOLOGICALS IS A PARTICLE-BASED AFFINITY CHROMATOGRAPHY PROCEDURE UTILIZING SEPHARAOSE-POLYMYXIN. ASIS TYPICAL FOR PARTICLE-BASED SEPARATIONS, THIS PROCESS IS SLOW AND INCONVENIENT DUE TO THE CHANGING CHARACTERISTICS OFAFFINITY COLUMNS WITH AGE. IN THIS PROJECT, STUDIES WILL BE DESIGNED TO OPTIMIZE SURFACE CHEMISTRIES FOR THE ATTACHMENT OF POLYMYXIN TO NON-PARTICLE-BASED SUPPORTS. THESE SUPPORTS ARE RIGID, HAVEEXCELLENT FLOW RATES, AND HAVE PHYSICAL PROPERTIES COMPATIBLE WITH HIGH-VOLUME APPLICATIONS, SUCH AS DETOXIFICATION OF PHARMACEUTICALS. ONCE THE ATTACHMENT CHEMISTRY IS DEFINED, THE OPTIMUM CONFIGURATION OF THE POLYMYXIN SUPPORT WILL BE DETERMINED IN COMBINATION WITH VACUUM ULTRAFILTRATION TO PROVIDE A SINGLE UNIT THAT WILL BOTH STERILIZE AND DETOXIFY IN A SINGLE STEP. IF SUCCESSFUL, THIS RESEARCH SHOULD LAY THE GROUNDWORK FOR THE DEVELOPMENT OF A PROTOTYPE DEVICE WHICH, WHEN REFINED, SHOULD FIND WIDESPREAD USES IN THE BIOTECHNOLOGY AND PHARMACEUTICAL INDUSTRIES. ADDITIONAL POSITIVE FEATURES OF SUCH A DEVICE ARE THAT IT WILL BE INEXPENSIVE, CONVENIENT TOUSE (PERHAPS EVEN REUSEABLE), AND AMENABLE TO INTERFACING WITH OTHER FLOW-THROUGH PROCESSES.