Abstract
The effect of an alternate strategy employing two different flowrates during loading was explored as a means of increasing system productivity in Protein-A chromatography. The effect of such a loading strategy was evaluated using a chromatographic model that was able to accurately predict experimental breakthrough curves for this Protein-A system. A gradient-based optimization routine is carried out to establish the optimal loading conditions (initial and final flowrates and switching time). The two-step loading strategy (using a higher flowrate during the initial stages followed by a lower flowrate) was evaluated for an Fc-fusion protein and was found to result in significant improvements in process throughput. In an extension of this optimization routine, dynamic loading capacity and productivity were simultaneously optimized using a weighted objective function, and this result was compared to that obtained with the single flowrate. Again, the dual-flowrate strategy was found to be superior.
Publication types
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Comparative Study
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Evaluation Study
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Validation Study
MeSH terms
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Adsorption
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Animals
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Antibodies, Monoclonal / chemistry
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Antibodies, Monoclonal / genetics
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Antibodies, Monoclonal / immunology
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Antibodies, Monoclonal / isolation & purification
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Antigen-Antibody Complex / chemistry
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Antigen-Antibody Complex / immunology
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Antigen-Antibody Complex / isolation & purification
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CHO Cells
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Chromatography, Affinity / methods*
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Complex Mixtures / chemistry
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Complex Mixtures / immunology
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Complex Mixtures / isolation & purification
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Computer Simulation
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Cricetinae
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Cricetulus
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Culture Media / chemistry
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Culture Media / isolation & purification
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Immunoglobulin Fc Fragments / chemistry*
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Immunoglobulin Fc Fragments / genetics
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Immunoglobulin Fc Fragments / immunology
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Immunoglobulin Fc Fragments / isolation & purification*
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Microfluidics / methods*
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Models, Chemical*
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Quality Control
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / immunology
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Recombinant Fusion Proteins / isolation & purification
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Staphylococcal Protein A / chemistry*
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Staphylococcal Protein A / immunology
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Staphylococcal Protein A / isolation & purification*
Substances
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Antibodies, Monoclonal
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Antigen-Antibody Complex
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Complex Mixtures
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Culture Media
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Immunoglobulin Fc Fragments
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Recombinant Fusion Proteins
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Staphylococcal Protein A