title: Application of coupled affinity-sizing chromatography for the detection of proteolyzed HSA-tagged proteins.
abstract: Coupled affinity liquid chromatography and size exclusion chromatography (ALC-SEC) is a technique that has been shown to successfully report product quality of proteins during cell expression and prior to the commencement of downstream processing chromatography steps. This method was applied to monitoring the degradation and subsequent partial remediation of a HSA-tagged protein which showed proteolysis, allowing for rapid cell line development to address this product quality dilemma. This paper outlines the novel application of this method for measuring and addressing protease-induced proteolysis.
authors.affiliation: Novartis Biologics Center, Novartis Institutes for Biomedical Research, Cambridge, MA, USA. Electronic address: anne.london@novartis.com.
authors.forename: Anne Serdakowski
authors.initials: AS
authors.lastname: London
authors.affiliation: Integrated Biologics Profiling, Novartis Pharmaceuticals, Cambridge, MA, USA.
authors.forename: Kunal
authors.initials: K
authors.lastname: Patel
authors.affiliation: Novartis Biologics Center, Novartis Institutes for Biomedical Research, Cambridge, MA, USA.
authors.forename: Lisa
authors.initials: L
authors.lastname: Quinn
authors.affiliation: Integrated Biologics Profiling, Novartis Pharmaceuticals, Cambridge, MA, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Lemmerer
mesh_terms: D000818:Animals; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel / Q000379:methods*; D003412:Cricetulus; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D011993:Recombinant Fusion Proteins
keywords: Affinity chromatography; Gel filtration chromatography; HPLC; Protease inhibitors; Purification