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Abstract
A real time monitoring of fouling in liquid chromatography has been presented. The versatility of the approach has been proven by successful implementation in three case studies with an error <1%. The first application demonstrates the monitoring of protein A ligand density and foulant concentration for assessing performance of protein A chromatography resin during purification of monoclonal antibodies. The observations have been supported from LC-MS/MS studies that were independently performed. The second application involves monitoring of foulant deposition during multimode cation exchange chromatography based purification of human serum albumin. Finally, in the third application, monitoring of foulants during multimodal hydrophobic interaction chromatography of recombinant human granulocyte colony stimulating factor is demonstrated. In all three cases, it is observed that the fluorescence intensity consistently increases with resin reuse as more foulants are deposited over time. The proposed approach can be readily used for real time monitoring of fouling and process control.
Highlights
Chromatographic separation continues to be the mainstay for purification of biotherapeutic products
We propose a robust fluorescence based approach to monitor fouling in process chromatography resin by presenting its use in three independent studies[7]
In view of the results, it is evident that the proposed approach can be readily used for real time monitoring of fouling and for process control
Summary
Chromatographic separation continues to be the mainstay for purification of biotherapeutic products. Current best practice is to use a combination of holistic indicators to monitor resin fouling These indicators include measurements of uniformity of column packing (plate height, asymmetry), product yield, and product quality. Researchers have demonstrated ATR-FTIR as a tool to monitor affinity chromatography purification of monoclonal antibody, but the measurement requires repacking of the resin and so cannot be directly applied to perform continuous monitoring of process scale chromatography[6]. In this communication, we propose a robust fluorescence based approach to monitor fouling in process chromatography resin by presenting its use in three independent studies[7]. In view of the results, it is evident that the proposed approach can be readily used for real time monitoring of fouling and for process control
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