Development of novel protein refolding using simulated moving bed chromatography

In vitro protein refolding is still one of the baffles in both structural biology and development of new biopharmaceuticals, especially for large-scale production of valuable proteins that are overexpressed as inclusion bodies inEscherichia coli. A new continuous refolding method using four zone simulated moving bed process based on size exclusion mechanism was developed to overcome difficulties of inclusion body refolding. Protein refolding using size exclusion SMB enables us to obtain refolded protein continuously with high productivity, low consumption of refolding buffer, and high efficiency of size exclusion medium. Thermodynamics and kinetic parameters for SMB operation were estimated from the best-fit values by comparing the simulation and experimental chromatography results. The SMB operation condition was obtained from the triangle theory, and experimental results were in good agreement with the simulation results