Characterization and validation of a middle-down hydrophobic interaction chromatography method to monitor methionine oxidation in IgG1
Post-translational modifications (PTMs) of therapeutic monoclonal antibodies (mAbs) can impact the efficacy of a drug. Methionine oxidation can alter the overall hydrophobicity of an antibody, thereby inducing conformational changes and affecting its biological activity. To ensure high quality, safe...
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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/S2949771X23000087 |
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| author | Somar Khalil Nisha Patel Francoise Bevillard-Kumar Cyrille Chéry William Burkitt John O’Hara Annick Gervais |
| author_facet | Somar Khalil Nisha Patel Francoise Bevillard-Kumar Cyrille Chéry William Burkitt John O’Hara Annick Gervais |
| author_sort | Somar Khalil |
| collection | DOAJ |
| description | Post-translational modifications (PTMs) of therapeutic monoclonal antibodies (mAbs) can impact the efficacy of a drug. Methionine oxidation can alter the overall hydrophobicity of an antibody, thereby inducing conformational changes and affecting its biological activity. To ensure high quality, safety, and efficacy of mAbs, routine monitoring of PTMs such as methionine (Met) oxidation is essential. Met oxidation in the fragment crystallizable (Fc) region of immunoglobulin-G1 (IgG1) is a potential critical quality attribute because it impacts not only the interaction with the neonatal Fc receptor and protein A but also the half-life of mAbs in serum circulation. Although bottom-up mass spectrometry provides high site specificity, it may have limited application in quality control workflows, and its complicated sample preparation could result in procedure-induced oxidation. In this study, we describe the development and characterization of a rapid and robust middle-down hydrophobic interaction chromatography method for monitoring Met oxidation in the Fc region of IgG1. Additionally, we assessed a comprehensive method validation package and demonstrated the specificity, linearity, precision, and accuracy of the new method within a range of 3.8–37.7%. The relative quantitative data provided by this method may be used in a regulated workflow to support process and formulation development as well as in the later stages of drug development and batch release and stability studies. |
| format | Article |
| id | doaj-art-a78f202ed3454585bd43fc9b3dfb7044 |
| 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-a78f202ed3454585bd43fc9b3dfb70442025-08-20T02:51:38ZengElsevierJournal of Pharmaceutical and Biomedical Analysis Open2949-771X2023-06-01110000810.1016/j.jpbao.2023.100008Characterization and validation of a middle-down hydrophobic interaction chromatography method to monitor methionine oxidation in IgG1Somar Khalil0Nisha Patel1Francoise Bevillard-Kumar2Cyrille Chéry3William Burkitt4John O’Hara5Annick Gervais6Department of Analytical Science Biologicals, UCB, Braine L’Alleud, Belgium; Correspondence to: Chemin du Foriest 1, Braine L’Alleud 1420, Belgium.Department of Biomolecular Formulation and Characterization Sciences, UCB, Slough, United KingdomDepartment of Analytical Science Biologicals, UCB, Braine L’Alleud, BelgiumDepartment of Analytical Science Biologicals, UCB, Braine L’Alleud, BelgiumDepartment of Biomolecular Formulation and Characterization Sciences, UCB, Slough, United KingdomDepartment of Biomolecular Formulation and Characterization Sciences, UCB, Slough, United KingdomDepartment of Analytical Science Biologicals, UCB, Braine L’Alleud, BelgiumPost-translational modifications (PTMs) of therapeutic monoclonal antibodies (mAbs) can impact the efficacy of a drug. Methionine oxidation can alter the overall hydrophobicity of an antibody, thereby inducing conformational changes and affecting its biological activity. To ensure high quality, safety, and efficacy of mAbs, routine monitoring of PTMs such as methionine (Met) oxidation is essential. Met oxidation in the fragment crystallizable (Fc) region of immunoglobulin-G1 (IgG1) is a potential critical quality attribute because it impacts not only the interaction with the neonatal Fc receptor and protein A but also the half-life of mAbs in serum circulation. Although bottom-up mass spectrometry provides high site specificity, it may have limited application in quality control workflows, and its complicated sample preparation could result in procedure-induced oxidation. In this study, we describe the development and characterization of a rapid and robust middle-down hydrophobic interaction chromatography method for monitoring Met oxidation in the Fc region of IgG1. Additionally, we assessed a comprehensive method validation package and demonstrated the specificity, linearity, precision, and accuracy of the new method within a range of 3.8–37.7%. The relative quantitative data provided by this method may be used in a regulated workflow to support process and formulation development as well as in the later stages of drug development and batch release and stability studies.http://www.sciencedirect.com/science/article/pii/S2949771X23000087Hydrophobic interactionsIdeS proteaseFragment crystallizable regionMethionine oxidationMethod developmentMonoclonal antibodies |
| spellingShingle | Somar Khalil Nisha Patel Francoise Bevillard-Kumar Cyrille Chéry William Burkitt John O’Hara Annick Gervais Characterization and validation of a middle-down hydrophobic interaction chromatography method to monitor methionine oxidation in IgG1 Journal of Pharmaceutical and Biomedical Analysis Open Hydrophobic interactions IdeS protease Fragment crystallizable region Methionine oxidation Method development Monoclonal antibodies |
| title | Characterization and validation of a middle-down hydrophobic interaction chromatography method to monitor methionine oxidation in IgG1 |
| title_full | Characterization and validation of a middle-down hydrophobic interaction chromatography method to monitor methionine oxidation in IgG1 |
| title_fullStr | Characterization and validation of a middle-down hydrophobic interaction chromatography method to monitor methionine oxidation in IgG1 |
| title_full_unstemmed | Characterization and validation of a middle-down hydrophobic interaction chromatography method to monitor methionine oxidation in IgG1 |
| title_short | Characterization and validation of a middle-down hydrophobic interaction chromatography method to monitor methionine oxidation in IgG1 |
| title_sort | characterization and validation of a middle down hydrophobic interaction chromatography method to monitor methionine oxidation in igg1 |
| topic | Hydrophobic interactions IdeS protease Fragment crystallizable region Methionine oxidation Method development Monoclonal antibodies |
| url | http://www.sciencedirect.com/science/article/pii/S2949771X23000087 |
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