Detection and quantitation of host cell proteins in monoclonal antibody drug products using automated sample preparation and data-independent acquisition LC-MS/MS
- Author:
Strasser LISA
1
;
Oliviero GIORGIO
;
Jakes CRAIG
;
Zaborowska IZABELA
;
Floris PATRICK
;
Meire Ribeiro da Silva
;
Füssl FLORIAN
;
Carillo SARA
;
Bones JONATHAN
Author Information
1. Characterization and Comparability Laboratory,NIBRT-National Institute for Bioprocessing Research and Training,Dublin,A94 X099,Ireland
- Keywords:
Data-independent acquisition;
Host cell proteins;
Critical quality attributes;
Liquid chromatography-mass spectrometry;
Monoclonal antibody;
Chinese hamster ovary cells
- From:
Journal of Pharmaceutical Analysis
2021;11(6):726-731
- CountryChina
- Language:Chinese
-
Abstract:
Ensuring the removal of host cell proteins (HCPs) during downstream processing of recombinant pro-teins such as monoclonal antibodies (mAbs) remains a challenge.Since residual HCPs might affect product stability or safety,constant monitoring is required to demonstrate their removal to be below the regulatory accepted level of 100 ng/mg.The current standard analytical approach for this procedure is based on ELISA;however,this approach only measures the overall HCP content.Therefore,the use of orthogonal methods,such as liquid chromatography-mass spectrometry (LC-MS),has been established,as it facilitates the quantitation of total HCPs as well as the identification and quantitation of the indi-vidual HCPs present.In the present study,a workflow for HCP detection and quantitation using an automated magnetic bead-based sample preparation,in combination with a data-independent acquisi-tion (DIA) LC-MS analysis,was established.Employing the same instrumental setup commonly used for peptide mapping analysis of mAbs allows for its quick and easy implementation into pre-existing workflows,avoiding the need for dedicated instrumentation or personnel.Thereby,quantitation of HCPs over a broad dynamic range was enabled to allow monitoring of problematic HCPs or to track changes upon altered bioprocessing conditions.
