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3D-LC-MS with D-2 multimethod option for fully automated assessment of multiple attributes of monoclonal antibodies directly from cell culture supernatants

Liesa Verscheure (UGent) , Gerd Vanhoenacker, Sonja Schneider, Tom Merchiers, Julie Storms, Patrick Sandra (UGent) , Frederic Lynen (UGent) and Koen Sandra (UGent)
(2022) ANALYTICAL CHEMISTRY. 94(17). p.6502-6511
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Organization
Abstract
Fully automated analysis of multiple structural attributes of monoclonal antibodies (mAbs) using three-dimensional liquid chromatography-mass spectrometry (3D-LC-MS) is described. The analyzer combines Protein A affinity chromatography in the first dimension (D-1) with a multimethod option in the second dimension (D-2) (choice between size exclusion (SEC), cation exchange (CEX), and hydrophobic interaction chromatography (HIC)) and desalting SEC-MS in the third dimension (D-3). This innovative 3D-LC-MS setup allows simultaneous and sequential assessment of mAb titer, size/charge/hydrophobic variants, molecular weight (MAN), amino acid (AA) sequence, and post-translational modifications (PTMs) directly from cell culture supernatants. The reported methodology that finds multiple uses throughout the biopharmaceutical development trajectory was successfully challenged by the analysis of different trastuzumab and tocilizumab samples originating from biosimilar development programs.
Keywords
2-DIMENSIONAL LIQUID-CHROMATOGRAPHY, EFFECTIVE PROTEIN SEPARATION, HYDROPHOBIC INTERACTION, THERAPEUTIC ANTIBODIES, PHASE CHROMATOGRAPHY, MASS-SPECTROMETRY, A CHROMATOGRAPHY, IDENTIFICATION, TRASTUZUMAB, TOCILIZUMAB

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MLA
Verscheure, Liesa, et al. “3D-LC-MS with D-2 Multimethod Option for Fully Automated Assessment of Multiple Attributes of Monoclonal Antibodies Directly from Cell Culture Supernatants.” ANALYTICAL CHEMISTRY, vol. 94, no. 17, 2022, pp. 6502–11, doi:10.1021/acs.analchem.1c05461.
APA
Verscheure, L., Vanhoenacker, G., Schneider, S., Merchiers, T., Storms, J., Sandra, P., … Sandra, K. (2022). 3D-LC-MS with D-2 multimethod option for fully automated assessment of multiple attributes of monoclonal antibodies directly from cell culture supernatants. ANALYTICAL CHEMISTRY, 94(17), 6502–6511. https://doi.org/10.1021/acs.analchem.1c05461
Chicago author-date
Verscheure, Liesa, Gerd Vanhoenacker, Sonja Schneider, Tom Merchiers, Julie Storms, Patrick Sandra, Frederic Lynen, and Koen Sandra. 2022. “3D-LC-MS with D-2 Multimethod Option for Fully Automated Assessment of Multiple Attributes of Monoclonal Antibodies Directly from Cell Culture Supernatants.” ANALYTICAL CHEMISTRY 94 (17): 6502–11. https://doi.org/10.1021/acs.analchem.1c05461.
Chicago author-date (all authors)
Verscheure, Liesa, Gerd Vanhoenacker, Sonja Schneider, Tom Merchiers, Julie Storms, Patrick Sandra, Frederic Lynen, and Koen Sandra. 2022. “3D-LC-MS with D-2 Multimethod Option for Fully Automated Assessment of Multiple Attributes of Monoclonal Antibodies Directly from Cell Culture Supernatants.” ANALYTICAL CHEMISTRY 94 (17): 6502–6511. doi:10.1021/acs.analchem.1c05461.
Vancouver
1.
Verscheure L, Vanhoenacker G, Schneider S, Merchiers T, Storms J, Sandra P, et al. 3D-LC-MS with D-2 multimethod option for fully automated assessment of multiple attributes of monoclonal antibodies directly from cell culture supernatants. ANALYTICAL CHEMISTRY. 2022;94(17):6502–11.
IEEE
[1]
L. Verscheure et al., “3D-LC-MS with D-2 multimethod option for fully automated assessment of multiple attributes of monoclonal antibodies directly from cell culture supernatants,” ANALYTICAL CHEMISTRY, vol. 94, no. 17, pp. 6502–6511, 2022.
@article{8758425,
  abstract     = {{Fully automated analysis of multiple structural attributes of monoclonal antibodies (mAbs) using three-dimensional liquid chromatography-mass spectrometry (3D-LC-MS) is described. The analyzer combines Protein A affinity chromatography in the first dimension (D-1) with a multimethod option in the second dimension (D-2) (choice between size exclusion (SEC), cation exchange (CEX), and hydrophobic interaction chromatography (HIC)) and desalting SEC-MS in the third dimension (D-3). This innovative 3D-LC-MS setup allows simultaneous and sequential assessment of mAb titer, size/charge/hydrophobic variants, molecular weight (MAN), amino acid (AA) sequence, and post-translational modifications (PTMs) directly from cell culture supernatants. The reported methodology that finds multiple uses throughout the biopharmaceutical development trajectory was successfully challenged by the analysis of different trastuzumab and tocilizumab samples originating from biosimilar development programs.}},
  author       = {{Verscheure, Liesa and Vanhoenacker, Gerd and Schneider, Sonja and Merchiers, Tom and Storms, Julie and Sandra, Patrick and Lynen, Frederic and Sandra, Koen}},
  issn         = {{0003-2700}},
  journal      = {{ANALYTICAL CHEMISTRY}},
  keywords     = {{2-DIMENSIONAL LIQUID-CHROMATOGRAPHY,EFFECTIVE PROTEIN SEPARATION,HYDROPHOBIC INTERACTION,THERAPEUTIC ANTIBODIES,PHASE CHROMATOGRAPHY,MASS-SPECTROMETRY,A CHROMATOGRAPHY,IDENTIFICATION,TRASTUZUMAB,TOCILIZUMAB}},
  language     = {{eng}},
  number       = {{17}},
  pages        = {{6502--6511}},
  title        = {{3D-LC-MS with D-2 multimethod option for fully automated assessment of multiple attributes of monoclonal antibodies directly from cell culture supernatants}},
  url          = {{http://doi.org/10.1021/acs.analchem.1c05461}},
  volume       = {{94}},
  year         = {{2022}},
}
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