Application of coupled affinity-sizing chromatography for the detection of proteolyzed HSA-tagged proteins

Anne Serdakowski London; Kunal Patel; Lisa Quinn; Martin Lemmerer

doi:10.1016/j.pep.2014.12.005

Application of coupled affinity-sizing chromatography for the detection of proteolyzed HSA-tagged proteins

Protein Expr Purif. 2015 Apr:108:80-84. doi: 10.1016/j.pep.2014.12.005. Epub 2014 Dec 13.

Authors

Anne Serdakowski London¹, Kunal Patel², Lisa Quinn³, Martin Lemmerer²

Affiliations

¹ Novartis Biologics Center, Novartis Institutes for Biomedical Research, Cambridge, MA, USA. Electronic address: anne.london@novartis.com.
² Integrated Biologics Profiling, Novartis Pharmaceuticals, Cambridge, MA, USA.
³ Novartis Biologics Center, Novartis Institutes for Biomedical Research, Cambridge, MA, USA.

PMID: 25514202
DOI: 10.1016/j.pep.2014.12.005

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.

Keywords: Affinity chromatography; Gel filtration chromatography; HPLC; Protease inhibitors; Purification.

MeSH terms

Animals
CHO Cells
Chromatography, Affinity / methods*
Chromatography, Gel / methods*
Cricetulus
Recombinant Fusion Proteins / chemistry
Recombinant Fusion Proteins / isolation & purification*

Substances

Recombinant Fusion Proteins