Characterization of Continuous Refolding Apparatus Using Different Mixing Methods.

Abstract

Taylor-eddy and paddle type apparatus were applied for refolding of lysozyme in a continuous system. The number of theoretical mixing stages was measured for both the apparatus and the effect of flow rate and mixing rate on the axial dispersion of solutions flowing inside of a ceramic membrane tube were investigated. Refolding at different concentrations of lysozyme was performed by addition of denatured lysozyme into refolding buffers flowing through the inside of the tube. For studying the effect of addition, different addition methods of denatured lysozyme into refolding buffer were devised by using ceramic membranes, from the total surface of the membrane tube and partially upside ceramic membrane. Comparing the efficiencies of continuous refolding with those of batch method, higher refolding efficiencies were obtained in the continuous system at the same concentrations of lysozyme and urea. Therefore, by combining the merits of the fed-batch method and continuous system, the refolding efficiency can be improved as well as throughput of refolding process.