title: Fluorescence based real time monitoring of fouling in process chromatography.
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.
authors.affiliation: Department of Chemical Engineering, Indian Institute of Technology, New Delhi, India.
authors.forename: Mili
authors.initials: M
authors.lastname: Pathak
authors.affiliation: Department of Biochemical Engineering, University College London, Torrington Place, London, UK.
authors.forename: Katherine
authors.initials: K
authors.lastname: Lintern
authors.affiliation: Department of Chemical Engineering, Indian Institute of Technology, New Delhi, India.
authors.forename: Viki
authors.initials: V
authors.lastname: Chopda
authors.affiliation: Department of Biochemical Engineering, University College London, Torrington Place, London, UK.
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Bracewell
authors.affiliation: Department of Chemical Engineering, Indian Institute of Technology, New Delhi, India.
authors.forename: Anurag S
authors.initials: AS
authors.lastname: Rathore