Abstract
Anisotropic polysulfone membranes were prepared with carboxypeptidase G1 embedded in the polymer structure. The enzymatically active flat and hollow-fiber membranes were obtained by precipitating the polymer from solution in an organic mixture in which an aqueous solution of the enzyme had been dispersed. The process has been found to be particularly suitable for the immobilization of enzymes in anisotropic hollow fibers that exhibited no detectable enzyme leakage upon perfusion. The pH profiles measured with the enzyme in free solution and in the embedded form were similar. Kinetic parameters of multitubular enzyme reactors were investigated by measuring the rate of hydrolysis of glutamate from folic acid or methotrexate at different flow rates and substrate concentrations. The relatively slow mass transfer in such reactors was found to affect strongly the observed kinetics. The results of in vitro experiments with 5000 fiber reactors suggest that hollow fiber cartridges prepared with such membranes have clinical potential for the extracorporeal removal of methotrexate from blood.
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References
Chang, T. M.S (ed.) (1977),Biomedical Applications of Immobilized Enzymes and Proteins, Vols. 1 and 2, Plenum Press, New York.
Messing, R. A. (ed.) (1975),Immobilized Enzymes for Industrial Reactors, Academic Press, New York.
Carr, P. W., and Bowers, L. D. (1980),Immobilized Enzymes in Analytical and Clinical Chemistry, Wiley-Interscience, New York.
Salmona, M., Saronio, C., Bartosek, I., Vecchi, A., and Mussini, I. (1974), Potential Biomedical Applications of Fiber-Entrapped L-Asparaginase and Urease, inlnsolubilized Enzymes, Salmona, M., Saronio, C., and Gerattini, S. (eds.), Ravan Press, New York, p. 189.
Pansolli, P., Giovenco, S., Dinelli, D., and Morisi, F. (1975), Kinetics of Fibre Entrapped Glucose Isomerase, inAnalysis and Control of Immobilized Enzyme Systems, Thomas, D., and Kernevez, J-P. (eds.), North Holland/American Elsevier, New York, p. 237.
Dinelli, D., Bartole, F., and Gulinelli, S. (1976), US Patent 3,947,325.
Marconi, W., Bartoli, F., Morisi, F., Pittalis, F., and Prosperi, G. (1979), Industrial Applications of Fibre-Entrapped Enzymes: Fibre-Entrapped Aspartase, inFEBS Trends in Enzymology, “Industrial and Clinical Enzymology,” Vitale, L. J., and Simeon, V., (eds.), Pergamon Press, Oxford, p. 49.
Gregor, H. P., and Rauf, P. W. (1975),Biotechnol. Bioeng. 17, 445.
Drioli, E., Gaeta, S., Carfagna, C., De Rosa, M., Gambarcorta, A., and Nicholaus, B. (1980),J. Membrane Sci. 6, 345.
Kohlwey, D. E., and Cheryan, M. (1980),Enzyme Microb. Technol. 3, 64.
Silman, R. W., Black, L. T., McGhee, J. T., and Bagley, E. B. (1980),Biotechnol. Bioeng. 22, 533.
Kalghatgi, K., Horváth, Cs., and Ambrus, CM. (1980),Res. Commun. Chem. Pathol. Pharmacol. 27, 55.
Rony, P. R. (1962),J. Amer. Chem. Soc. 94, 8247.
Rony, P. R. (1971),Biotechnol. Bioeng. 12, 431.
Waterland, L. R., Michaelis, A. S., and Robertson, C. R. (1974),AIChEJ. 20, 50.
Lewis, W., and Middleman, S. (1974),AIChEJ. 20, 1012.
Mashelkar, R. A., and Ramachandran, P. A. (1975),J. Appl. Chem. Biotechnol. 25, 867.
Amicon Corporation, Lexington, MA, (1979), Diafilters, Publication # 1435.
Henderson, L. W., Ford, C., Colton, C. K., Blumley, L. W., and Bixter, H. J. (1970),Amer. Soc. Artif. Organs 16, 107.
Bertino, J. R., Condos, S., Horváth, Cs., Kalghatgi, K., and Pedersen, H. (1978),Cancer Res. 38, 1936.
Ambrus, C. M., Ambrus, J. L., Horváth, Cs., Pedersen, H., Sharma, S., Kant, C., Mirand, E., Guthrie, R., and Paul, T. (1978),Science 201, 837.
Chick, W. L., Like, A. A., Lauris, V., Galetti, P. M., Richardson, P. D., Panol, G., Mix, T. W., and Colton, C. K. (1975),Trans. Amer. Soc. Artif. Organs 21, 8.
McCullough, J. L., Chabner, B. A., and Bertino, J. R. (1971),J. Biol. Chem. 246, 7207.
Kalghatgi, K., and Bertino, J. R. (1981), Folate Degrading Enzymes: A review with special emphasis on Carboxypeptidase G1, inEnzymes as Drugs, Holcenberg, J. C., and Roberts, J. (eds.), Wiley, New York, p. 77.
Bertino, J. R., and Fischer, G. A. (1964),Methods Med. Res. 10, 297.
Kalghatgi, K., and Horváth, Cs., unpublished results.
Michaels, A. S. (1968),Chem. Eng. Progr. 64, 131.
Michaels, A. S. (1971), US Patent #3,615,024.
Cross, R. A. (1972), US Patent #3,691,068.
Cross, R. A. (1972a),AlChE Symp. Ser. 68, 15.
Weber, K., and Osborn, M. (1969),J. Biol. Chem. 244, 4406.
Kohman, G. T., and McAfee, K. B. (1962), US Patent #3,019,853.
Horváth, Cs., and Solomon, B. A. (1972),Biotechnol. Bioeng. 14, 885.
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Pitt, A.M., Cramer, S.M., Czernicki, A.B. et al. Anisotropic membranes with carboxypeptidase G1. Appl Biochem Biotechnol 8, 55–68 (1983). https://doi.org/10.1007/BF02798348
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DOI: https://doi.org/10.1007/BF02798348