Optimization of preparative ion-exchange chromatography of proteins: linear gradient separations

Analysis of steric mass-action model for protein adsorption equilibrium onto porous anion-exchange adsorbent

“…To date, this model has been widely used to describe protein adsorption equilibrium. These reported results demonstrated the efficacy of the SMA model in predicting the nonlinear adsorption behavior of multicomponent protein in step gradient (Chen et al, 2003b), linear gradient (Gallant et al, 1996;Jakobssona et al, 2005) and displacement ion-exchange systems (Natarajan et al, 2000) as well as metal affinity systems (Vunnum et al, 1995).…”

“…In their study, the values of and a of BSA in pH 7.8 Tris-HCl buffer were 4.3 and 142, respectively. Gallant et al (1996) reported that the values of K a , and a for adsorption of lysozyme onto a strong cation exchange column (SCX) (sulfopropyl, 54 × 5 mm I.D.) with a column capacity of 525 mmol/l in pH 6.0 phosphate buffer were 0.211, 6.14 and 0, respectively.…”

“…In order to keep operating costs low in large-scale separation processes, the optimization of chromatographic operation becomes increasingly important (Chen et al, 2003a). It is well known that the gradient elution by increasing ionic strength is widely adopted in the ion-exchange chromatography because the adsorption of most proteins decreases with increasing ionic strength (Gallant et al, 1996;Lewus and Carta, 1999). Therefore, to simulate the elution process, it is necessary to take into account the influence of ionic strength on the ion-exchange equilibrium.…”

Use and Optimization of a Dual-Flowrate Loading Strategy To Maximize Throughput in Protein-A Affinity Chromatography

“…In that case, total time taken for loading can be calculated as Productivity in that case will be defined as Optimization. To accomplish any optimization in preparative chromatographic systems, one needs to consider four different factors: objective function, parameters, decision variables and constraints (17). Optimization consists of maximizing or minimizing a given objective function.…”

“…Luo and Hsu (15) used an iterative scheme to optimize the gradient slope in linear ion-exchange chromatography. In addition, iterative optimization schemes have been used for optimizing step gradient (16), linear gradient (17) and displacement systems (18). Suwondo et al (19) employed a generalized reduced gradient method to maximize throughput in ideal linear chromatography.…”