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Dual Data-Independent Acquisition Approach Combining Global HCP Profiling and Absolute Quantification of Key Impurities during Bioprocess Development

Husson, G; Delangle, A; O'Hara, J; Cianférani, S; Gervais, A; Van Dorsselaer, A; Bracewell, DG; (2017) Dual Data-Independent Acquisition Approach Combining Global HCP Profiling and Absolute Quantification of Key Impurities during Bioprocess Development. Analytical Chemistry , 90 (2) pp. 1241-1247. 10.1021/acs.analchem.7b03965. Green open access

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Abstract

Host cell proteins (HCP) are a major class of impurities derived from recombinant protein production process. While HCP are usually monitored by ELISA, mass spectrometry (MS) based approaches are emerging as promising orthogonal methods. Here, we developed an original method relying on data independent acquisition (DIA) coupling global HCP amounts estimation (Top 3) and absolute quantification with isotope dilution (ID). The method named Top 3-ID-DIA was benchmarked against ELISA and a gold standard selected reaction monitoring assay (ID-SRM). Various samples generated at different steps and conditions of the purification process, including different culture durations, harvest procedures and purification protocols were used to compare the methods. Overall, HCP were quantified over 5 orders of magnitude and down to sub-ppm level. The Top 3-ID-DIA strategy proved to be equivalent to the gold standard ID-SRM in terms of sensitivity (1-10 ppm), accuracy and precision. Moreover, 81% of the Top 3 estimations were accurate within a factor of 2 when compared to ID-SRM. Thus, our approach aggregates global HCP profiling for comprehensive process understanding with absolute quantification of key HCP within a single analysis, and provides an improved support for bioprocess development and product purity assessment.

Type: Article
Title: Dual Data-Independent Acquisition Approach Combining Global HCP Profiling and Absolute Quantification of Key Impurities during Bioprocess Development
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1021/acs.analchem.7b03965
Publisher version: http://doi.org/10.1021/acs.analchem.7b03965
Language: English
Additional information: © 2017 American Chemical Society. This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions.
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/10040612
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