Improving affinity chromatography resin efficiency using semi-continuous chromatography
| Autor: | Anupa M. George, Ekta Mahajan, Bradley Wolk |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Chromatography
Continuous operation Chemistry Organic Chemistry Antibodies Monoclonal General Medicine CHO Cells Raw material Biochemistry Column (database) Chromatography Affinity Analytical Chemistry Resins Synthetic Countercurrent chromatography Cricetulus Laboratory Chemicals Affinity chromatography Volume (thermodynamics) Cricetinae Batch processing Equipment Reuse Animals Chromatography column Staphylococcal Protein A |
| Zdroj: | Journal of chromatography. A. 1227 |
| ISSN: | 1873-3778 |
| Popis: | Protein A affinity chromatography is widely used for purification of monoclonal antibodies (MAbs) from harvested cell culture fluid (HCCF). At the manufacturing scale, the HCCF is typically loaded on a single Protein A affinity chromatography column in cycles until all of the HCCF is processed. Protein A resin costs are significant, comprising a substantial portion of the raw material costs in MAb manufacturing. Cost can be reduced by operating the process continuously using multiple smaller columns to a higher binding capacity in lieu of one industrial scale column. In this study, a series of experiments were performed using three 1-ml Hi-Trap™ MabSelect SuRe™ columns on a modified AKTA™ system operated according to the three Column Periodic Counter Current Chromatography (3C PCC) principle. The columns were loaded individually at different times until the 70% breakthrough point was achieved. The HCCF with unbound protein from the column was then loaded onto the next column to capture the MAb, preventing any protein loss. At any given point, all three columns were in operation, either loading or washing, enabling a reduction in processing time. The product yield and quality were evaluated and compared with a batch process to determine the effect of using the three column continuous process. The continuous operation shows the potential to reduce both resin volume and buffer consumption by ∼40%, however the system hardware and the process is more complex than the batch process. Alternative methods using a single standard affinity column, such as recycling load effluent back to the tank or increasing residence time, were also evaluated to improve Protein A resin efficiency. These alternative methods showed similar cost benefits but required longer processing time. |
| Databáze: | OpenAIRE |
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