Fast determination of conditions for maximum dynamic capacity in cation-exchange chromatography of human monoclonal antibodies

Faude, Alexander; Zacher, Dörthe; Müller, Egbert; Böttinger, Heiner

doi:10.1016/j.chroma.2007.03.114

Cited by 31 publications

(18 citation statements)

References 38 publications

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“…15 mS/cm). Lowering the conductivity to 5 mS/cm, the isoionic point (pH 6.4) shifts closer to the isoelectric point, which correlates with the results of Faude et al (2007). Comparing the AUC results of the SE-HPLC at different pH values, the dimerization ratio could be identified (see Fig.…”

Section: Scfv Hu225 Characteristics and Quantificationsupporting

confidence: 72%

Process development of periplasmatically produced single chain fragment variable against epidermal growth factor receptor in Escherichia coli

Lindner

Moosmann

Dietrich

et al. 2014

Journal of Biotechnology

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Section: Scfv Hu225 Characteristics and Quantificationsupporting

confidence: 72%

Process development of periplasmatically produced single chain fragment variable against epidermal growth factor receptor in Escherichia coli

Lindner

Moosmann

Dietrich

et al. 2014

Journal of Biotechnology

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“…Similar results were reported by Faude et al (2007) for the dynamic-binding capacity of two different monoclonal antibodies on a series of cation exchange resins (Fractogel EMD, SP Sepharose XL, and Toyopearl Prototype S). In all cases, there was a distinct maximum in the dynamic-binding capacity at intermediate pH (or conductivity), with the location of this maximum well correlated with the protein zeta potential.…”

Section: Introductionsupporting

confidence: 87%

“…It completely neglects any electrokinetic effects, it ignores the possibility of surface diffusion and any change in effective pore size due to collapse of the grafted dextran, and the approach used to evaluate the dynamic-binding capacity is unable to account for the detailed breakthrough behavior or the passage of the concentration front through the chromatographic column. However, the good agreement between the model calculations and experimental data provides a strong indication that the steric/electrostatic phenomenon, which was the primary focus of this theoretical analysis, provides the dominant mechanism for the unusual chromatographic behavior observed by Harinarayan et al (2006) and Faude et al (2007) for ion exchange of antibodies. In addition, this model framework may provide a means for estimating optimal conditions for ion exchange chromatography based on experimental measurements of the resin pore size distribution and the equilibrium (static) binding capacity along with an appropriate correlation for b, which describes the contribution of the resin charge to the effective surface charge density of the pore wall after protein binding.…”

Section: Discussionmentioning

confidence: 79%

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Modeling electrostatic exclusion effects during ion exchange chromatography of monoclonal antibodies

Zydney

Harinarayan²,

Reis³

2008

Biotech & Bioengineering

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Recent experimental studies have shown a reduction in dynamic-binding capacity for both monoclonal antibodies and antigen-binding fragments at very low conductivity, conditions that should generate the greatest electrostatic attraction. This behavior has been attributed to the steric and electrostatic exclusion of the charged protein from the entrance of the resin pores. This manuscript presents a quantitative mathematical description of this phenomenon. The protein partition coefficient was evaluated using models for the partitioning of a charged sphere into a charged cylindrical pore, with the pore size distribution evaluated by inverse size exclusion chromatography. The results were in very good agreement with experimental data for batch protein uptake and dynamic-binding capacity over a range of pH and conductivity. This theoretical framework provides important insights into the behavior of ion exchange chromatography for protein purification.

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“…The maximum product binding capacity is important to enable economic processing (13)(14)(15). The experimental conditions affecting the observed capacity of an ion exchange resin are: pH, ionic strength of the buffer, nature of the counter-ion, flow rate, and temperature (2,7,8,(16)(17)(18)(19)(20). Flow rate is important for dynamic binding capacity.…”

Section: Introductionmentioning

confidence: 99%

Optimization of Dynamic Binding Capacity of Anion Exchange Chromatography Media for Recombinant Erythropoietin Purification

et al. 2014

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Background:The dynamic binding capacity (DBC) of a chromatography matrix in protein purification is the amount of the total protein absorbed into the matrix, before occurrence of a significant break in the breakthrough curve. Optimization of the process criteria for maximum DBC avoids extra process scale-up and reduces the processing time, costs and protein loss. Taguchi method is a simple useful tool in experimental design to estimate the optimal condition with minimum experiments. Objectives: In this research, linear flow rate, pH and protein concentration of the feed were checked according to an L9 orthogonal Taguchi array, to estimate the best conditions for maximum DBC of Q-sepharose fast flow (QSFF) resin in recombinant human erythropoietin purification process. Materials and Methods: A crud sample containing human recombinant erythropoietin was harvested from a cell culture of Chinese hamster ovary (CHO) cell line. Desalted harvests with different total protein concentrations (30, 40 and 50 µg.mL -1 ) and pH values (5, 6 and 7) were loaded into a packed column of QSFF with different linear flow rates (60, 120 and 280 cm.h -1 ) up to 10% of the breakthrough curve. The total protein loading to the column was checked by UV absorbance and Lowry method, and erythropoietin concentration was measured by ELISA. Analysis of variance (ANOVA) was applied to determine the optimum condition. Results: Finally, total protein concentration of 50 µg.mL -1 , pH of 5 and flow rate of 120 cm.h -1 , were anticipated as the optimal process conditions with 5.85 mg.mL -1 of resin as the dynamic binding capacity. Conclusions:Experiments with anticipated optimal criteria were performed three times and no significant difference was observed (p = 0.136, and 6.06 mg/mL as the average dynamic binding capacity).

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Fast determination of conditions for maximum dynamic capacity in cation-exchange chromatography of human monoclonal antibodies

Cited by 31 publications

References 38 publications

Process development of periplasmatically produced single chain fragment variable against epidermal growth factor receptor in Escherichia coli

Process development of periplasmatically produced single chain fragment variable against epidermal growth factor receptor in Escherichia coli

Modeling electrostatic exclusion effects during ion exchange chromatography of monoclonal antibodies

Optimization of Dynamic Binding Capacity of Anion Exchange Chromatography Media for Recombinant Erythropoietin Purification

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