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Quantitative definition and monitoring of the host cell protein proteome using iTRAQ: a study of an industrial mAb producing CHO-S cell line

Chiverton, Lesley M., Evans, Caroline, Pandhal, Jagroop, Landels, Andrew R., Rees, Byron J., Levison, Peter R., Wright, Phillip C., Smales, Christopher Mark (2016) Quantitative definition and monitoring of the host cell protein proteome using iTRAQ: a study of an industrial mAb producing CHO-S cell line. Biotechnology Journal, 11 (8). pp. 1014-1024. ISSN 1860-6768. (doi:10.1002/biot.201500550)

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Abstract

There are few studies defining CHO host cell proteins (HCPs) and the flux of these throughout a downstream purification process. Here we have applied quantitative iTRAQ proteomics to follow the HCP profile of an antibody (mAb) producing CHO-S cell line throughout a standard downstream purification procedure consisting of a Protein A, cation and anion exchange process. We used both 6 sample iTRAQ experiment to analyze technical replicates of three samples, which were culture harvest (HCCF), Protein A flow through and Protein A eluate and an 8 sample format to analyze technical replicates of four sample types; HCCF compared to Protein A eluate and subsequent cation and anion exchange purification. In the 6 sample iTRAQ experiment, 8781 spectra were confidently matched to peptides from 819 proteins (including the mAb chains). Across both the 6 and 8 sample experiments 936 proteins were identified. In the 8 sample comparison, 4187 spectra were confidently matched to peptides from 219 proteins. We then used the iTRAQ data to enable estimation of the relative change of individual proteins across the purification steps. These data provide the basis for application of iTRAQ for process development based upon knowledge of critical HCPs.

Item Type: Article
DOI/Identification number: 10.1002/biot.201500550
Uncontrolled keywords: Bioprocessing; Chinese hamster ovary (CHO); Host cell proteins; iTRAQ; Monoclonal antibody
Subjects: Q Science > Q Science (General)
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Mark Smales
Date Deposited: 12 Aug 2016 15:31 UTC
Last Modified: 29 May 2019 17:42 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/56829 (The current URI for this page, for reference purposes)
Smales, Christopher Mark: https://orcid.org/0000-0002-2762-4724
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