Refolding of proteins in a CSTR

A model to predict refolding of proteins in a continuous stirred tank reactor (CSTR) was developed and compared to a batch refolding process with simple dilution of the protein in a stirred tank reactor. For experimental verification of the model a continuous refolding of a model protein (αα-lactalbumin) was performed in a CSTR. The refolding process of denatured and fully reduced αα-lactalbumin could be accurately predicted by a set of differential equations assuming a first order reaction rate for folding and a second order reaction rate for aggregation. The system composed of a CSTR with an additional diafiltration circuit for removal of denaturing agents from the feed solution and to maintain constant refolding conditions. Based on the folding kinetic the dynamic behavior of such a continuous refolding reactor was simulated under different operating conditions. It was shown that the refolding efficiency was higher compared to batch dilution under certain conditions, namely high residence times. The yield of refolded protein could further increased by recycling the outlet stream containing unfolded protein to the reactor entrance.