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  • Biochemistry and Biotechnology  (2)
  • Wiley-Blackwell  (2)
  • American Physical Society
  • Molecular Diversity Preservation International
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  • Wiley-Blackwell  (2)
  • American Physical Society
  • Molecular Diversity Preservation International
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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 42 (1993), S. 1252-1262 
    ISSN: 0006-3592
    Keywords: biomedical reactor ; extracorporeal circuit ; hypercholesteremia New Zealand white rabbits ; immobilized phospholipase A2 ; plasma separator reactor (PSR) ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The purpose of this study was to design a biomedical reactor that reduces plasma cholesterol when incorporated in an in vivo extracorporeal system. Phospholipase A2, immobilized onto Agarose beads and housed inside the bioreactor, modifies plasma low density lipoprotein (LDL) into a form that is rapidly removed from circulation. In a packed bed reactor, the enzymatic conversion of LDL to the modified form (with plasma taken from hypercholesterolemic New Zealand white rabbits) was relatively low, 25% ± 6 for a single pass of plasma through the reactor. An extended bed reactor, a hybrid of fluidized and packed bed reactors, was then developed to increase the conversion. This reactor displays a single pass conversion of 60% ± 5 under optimal flow conditions. An evaluation of the flow rate through the reactor indicates that the system is limited by external mass transfer when employed under in vivo conditions. In addition, this system requires blood separation before the enzyme modification, which complicates the circuit control. Therefore, a new system was designed for in vivo use with rabbits. The resulting design, called the plasma separator reactor (PSR), combines plasma separation and enzymatic conversion in a single chamber. The PSR has three advantages over other studied systems: improved external mass transfer conditions, easy controlability, and simple set-up procedures. Single pass conversion reached 52% ± 12 in suboptimal flow under simulated in vivo conditions. This reactor was also tested in vivo with hypercholesterolemic New Zealand white rabbits. A continuous conversion of up to 80% ± 6 of rabbit plasma phospholipids was observed during 90 min of blood circulation (5 mL/min). The decrease in total plasma cholesterol reached a level of 60% of the initial value and was observed to be a function of the bioreactor enzyme activity. © 1993 John Wiley & Sons, Inc.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 0006-3592
    Keywords: hollow fiber ; bioreactor ; immobilized enzymes ; porosity ; phospholipase A2 ; low densitylipoprotein ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Previous studies have shown that the modification of low density lipoprotein (LDL) by the enzyme phospholipase A2(PLA2)results in a reduction of cholesterol levels in the plasma of hypercholesterolemic rabbits, due to accelerated clearance of the modified LDL. In the current study, we established techniques and optimized the ratio of enzyme to support for the immobilization of PLA2 on a polymeric support. Hollow fiber bioreactors made from polytetrafluoroethylene (PTFE) polymers were used to encapsulate immobilized PLA2. This design was adopted to eliminate hemolysis of red blood cells by the enzyme. Characterization of the resulting immobilized enzyme in terms of its activity, Michaelis-Menten kinetic constants, and the variation of its activity with incubation time is presented. The enzyme activity was not significantly altered upon incubation at 37°C in lipoprotein-deficient serum (LPDS), over the course of 2 months. The Michaelis-Menten kinetics constants are KM = 8.9 mM, Vmax = 6434.2 for the free enzyme and KappM = 16.7 mM, Vappmax = 619.7 for the immobilized enzyme. These data suggest that a system based on immobilized PLA2 in conjunction with hollow fiber bioreactors (HFBs) may be a good candidate for lowering LDL levels in plasma. © 1995 John Wiley & Sons, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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