Label-free shotgun proteomics: Exploiting a reliable and sensitive method to monitor residual host-cell proteins in monoclonal antibody products
The removal of host cell proteins (HCPs) remains a challenge in the downstream processing of monoclonal antibodies (mAbs). It is critical to monitor residual HCPs since they can have an impact on product stability and safety. The enzyme-linked immunosorbent assay (ELISA) only quantifies the total am...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-06-01
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| Series: | Journal of Pharmaceutical and Biomedical Analysis Open |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2949771X23000129 |
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| author | Somar Khalil Adeline Wychowski Cyrille Chéry Annick Gervais |
| author_facet | Somar Khalil Adeline Wychowski Cyrille Chéry Annick Gervais |
| author_sort | Somar Khalil |
| collection | DOAJ |
| description | The removal of host cell proteins (HCPs) remains a challenge in the downstream processing of monoclonal antibodies (mAbs). It is critical to monitor residual HCPs since they can have an impact on product stability and safety. The enzyme-linked immunosorbent assay (ELISA) only quantifies the total amount of HCPs and does not indicate the identity or quantity of any specific HCP. Mass spectrometry-based proteomics applications demonstrate an advantage in quantitatively profiling individual HCPs. However, technical reproducibility, dynamic range, and ensuring an acceptable statistical significance of scoring measurements are critical hurdles that ought to be overcome. In this paper, we describe a sensitive and reproducible shotgun proteomics approach that includes an efficient mAb depletion strategy and addresses the shortcomings of such workflows. Our methodology is potentially a valuable complement or alternative to ELISA. Additionally, it can facilitate the purification process development and the evaluation of ELISA kits upon process changes. |
| format | Article |
| id | doaj-art-9b379f6a9d204c34a5dc595302f2fa39 |
| institution | DOAJ |
| issn | 2949-771X |
| language | English |
| publishDate | 2023-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Pharmaceutical and Biomedical Analysis Open |
| spelling | doaj-art-9b379f6a9d204c34a5dc595302f2fa392025-08-20T02:51:38ZengElsevierJournal of Pharmaceutical and Biomedical Analysis Open2949-771X2023-06-01110001210.1016/j.jpbao.2023.100012Label-free shotgun proteomics: Exploiting a reliable and sensitive method to monitor residual host-cell proteins in monoclonal antibody productsSomar Khalil0Adeline Wychowski1Cyrille Chéry2Annick Gervais3Correspondence to: Chemin du Foriest 1, Braine L’Alleud 1420, Belgium.; Department of Analytical Science Biologicals, UCB, Braine L’Alleud, BelgiumDepartment of Analytical Science Biologicals, UCB, Braine L’Alleud, BelgiumDepartment of Analytical Science Biologicals, UCB, Braine L’Alleud, BelgiumDepartment of Analytical Science Biologicals, UCB, Braine L’Alleud, BelgiumThe removal of host cell proteins (HCPs) remains a challenge in the downstream processing of monoclonal antibodies (mAbs). It is critical to monitor residual HCPs since they can have an impact on product stability and safety. The enzyme-linked immunosorbent assay (ELISA) only quantifies the total amount of HCPs and does not indicate the identity or quantity of any specific HCP. Mass spectrometry-based proteomics applications demonstrate an advantage in quantitatively profiling individual HCPs. However, technical reproducibility, dynamic range, and ensuring an acceptable statistical significance of scoring measurements are critical hurdles that ought to be overcome. In this paper, we describe a sensitive and reproducible shotgun proteomics approach that includes an efficient mAb depletion strategy and addresses the shortcomings of such workflows. Our methodology is potentially a valuable complement or alternative to ELISA. Additionally, it can facilitate the purification process development and the evaluation of ELISA kits upon process changes.http://www.sciencedirect.com/science/article/pii/S2949771X23000129Affinity chromatographyDownstream processingHost cell proteinsLabel freeMass spectrometryMonoclonal antibody |
| spellingShingle | Somar Khalil Adeline Wychowski Cyrille Chéry Annick Gervais Label-free shotgun proteomics: Exploiting a reliable and sensitive method to monitor residual host-cell proteins in monoclonal antibody products Journal of Pharmaceutical and Biomedical Analysis Open Affinity chromatography Downstream processing Host cell proteins Label free Mass spectrometry Monoclonal antibody |
| title | Label-free shotgun proteomics: Exploiting a reliable and sensitive method to monitor residual host-cell proteins in monoclonal antibody products |
| title_full | Label-free shotgun proteomics: Exploiting a reliable and sensitive method to monitor residual host-cell proteins in monoclonal antibody products |
| title_fullStr | Label-free shotgun proteomics: Exploiting a reliable and sensitive method to monitor residual host-cell proteins in monoclonal antibody products |
| title_full_unstemmed | Label-free shotgun proteomics: Exploiting a reliable and sensitive method to monitor residual host-cell proteins in monoclonal antibody products |
| title_short | Label-free shotgun proteomics: Exploiting a reliable and sensitive method to monitor residual host-cell proteins in monoclonal antibody products |
| title_sort | label free shotgun proteomics exploiting a reliable and sensitive method to monitor residual host cell proteins in monoclonal antibody products |
| topic | Affinity chromatography Downstream processing Host cell proteins Label free Mass spectrometry Monoclonal antibody |
| url | http://www.sciencedirect.com/science/article/pii/S2949771X23000129 |
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