Design and optimization of protein refolding with crossflow ultrafiltration

Buffer recycling in downstream processing of biologics

“…This technique allows refolding at high protein concentrations which reduces buffer consumption. Ryś et al [14] successfully demonstrated with a crossflow diafiltration unit that permeate recycling can further decrease buffer consumption ( Figure 1a). By applying diafiltration, Schlegl et al [15] assured constant refolding conditions in a CSTR.…”

“…The simplest but also most time consuming method is by batch dilution [8,9 ,10 ,11-13]. Strategies to improve refolding productivity and efficiency include crossflow ultrafiltration processes [14], continuous stirred tank reactors (CSTR) [15], and annular [16,17] and simulated moving bed chromatography [18 ]. Intensification of these processes has 2 Biotechnology and bioprocess engineering been achieved by different recycling strategies.…”

Advances in monitoring and control of refolding kinetics combining PAT and modeling

“…The mechanism behind the folding of proteins is of great interest and many research groups tried to describe it using a variety of models (Cleland et al 1992 ; Dong et al 2004 ; Jungbauer and Kaar 2007 ; Kiefhaber et al 1991 ; Zettlmeissl et al 1979 ; Ryś et al 2015a ). The general structure of these models resembles one of the three classical models shown by Dill and Chan (Dill and Chan 1997 ), namely…”

“…In addition, this may result in aggregate formation due to prolonged protein exposure at medium denaturant concentration (Cabrita and Bottomley 2004 ; Tsumoto et al 2003 ). Diafiltration is shown to be an efficient system for protein refolding, not only to achieve higher yields than refolding in batch mode, but by reducing buffer consumption as well (Ryś et al 2015a ). However, fouling of membranes by aggregated proteins poses a problem (De Bernardez Clark 1998 ; Ryś et al 2015a ).…”

“…Diafiltration is shown to be an efficient system for protein refolding, not only to achieve higher yields than refolding in batch mode, but by reducing buffer consumption as well (Ryś et al 2015a ). However, fouling of membranes by aggregated proteins poses a problem (De Bernardez Clark 1998 ; Ryś et al 2015a ). The applied pressure in the high-pressure refolding method dissociates the existing protein aggregates and prevents the formation of further aggregates while requiring only small concentrations of chaotropic substances.…”