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High-Throughput Biophysical Analysis and Data Visualization of Conformational Stability of an IgG1 Monoclonal Antibody (mAb) After Deglycosylation
dc.contributor.author | Alsenaidy, Mohammad A. | |
dc.contributor.author | Kim, Jae Hyun | |
dc.contributor.author | Majumdar, Ranajoy | |
dc.contributor.author | Weis, David D. | |
dc.contributor.author | Joshi, Sangeeta B. | |
dc.contributor.author | Tolbert, Thomas J. | |
dc.contributor.author | Middaugh, C. Russell | |
dc.contributor.author | Volkin, David B. | |
dc.date.accessioned | 2017-04-25T19:03:41Z | |
dc.date.available | 2017-04-25T19:03:41Z | |
dc.date.issued | 2013-11 | |
dc.identifier.citation | Alsenaidy, M. A., Kim, J. H., Majumdar, R., Weis, D. D., Joshi, S. B., Tolbert, T. J., … Volkin, D. B. (2013). High-Throughput Biophysical Analysis and Data Visualization of Conformational Stability of an IgG1 Monoclonal Antibody (mAb) After Deglycosylation. Journal of Pharmaceutical Sciences, 102(11), 10.1002/jps.23730. http://doi.org/10.1002/jps.23730 | en_US |
dc.identifier.uri | http://hdl.handle.net/1808/23769 | |
dc.description.abstract | The structural integrity and conformational stability of an IgG1 monoclonal antibody (mAb), after partial and complete enzymatic removal of the N-linked Fc glycan, was compared to the untreated mAb over a wide range of temperature (10° to 90°C) and solution pH (3 to 8) using circular dichroism, fluorescence spectroscopy, and static light scattering combined with data visualization employing empirical phase diagrams (EPDs). Subtle to larger stability differences between the different glycoforms were observed. Improved detection of physical stability differences was then demonstrated over narrower pH range (4.0-6.0) using smaller temperature increments, especially when combined with an alternative data visualization method (radar plots). Differential scanning calorimetry and differential scanning fluorimetry were then utilized and also showed an improved ability to detect differences in mAb glycoform physical stability. Based on these results, a two step methodology was used in which mAb glycoform conformational stability is first screened with a wide variety of instruments and environmental stresses, followed by a second evaluation with optimally sensitive experimental conditions, analytical techniques and data visualization methods. With this approach, high-throughput biophysical analysis to assess relatively subtle conformational stability differences in protein glycoforms is demonstrated. | en_US |
dc.publisher | Elsevier | en_US |
dc.rights | This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License 4.0 (CC BY-NC-ND 4.0), which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. | en_US |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | en_US |
dc.subject | Glycosylation | en_US |
dc.subject | Monoclonal Antibody | en_US |
dc.subject | Stability | en_US |
dc.subject | Structure | en_US |
dc.subject | Conformation | en_US |
dc.subject | Biophysical | en_US |
dc.subject | Formulation | en_US |
dc.title | High-Throughput Biophysical Analysis and Data Visualization of Conformational Stability of an IgG1 Monoclonal Antibody (mAb) After Deglycosylation | en_US |
dc.type | Article | en_US |
kusw.kuauthor | Alsenaidy, Mohammad A. | |
kusw.kuauthor | Kim, Jae Hyun | |
kusw.kuauthor | Majumdar, Ranajoy | |
kusw.kuauthor | Weis, David D. | |
kusw.kuauthor | Joshi, Sangeeta B. | |
kusw.kuauthor | Tolbert, Thomas J. | |
kusw.kuauthor | Middaugh, C. Russell | |
kusw.kuauthor | Volkin, David B. | |
kusw.kudepartment | Pharmaceutical Chemistry | en_US |
dc.identifier.doi | 10.1002/jps.23730 | en_US |
kusw.oaversion | Scholarly/refereed, author accepted manuscript | en_US |
kusw.oapolicy | This item meets KU Open Access policy criteria. | en_US |
dc.identifier.pmid | PMC3832129 | en_US |
dc.rights.accessrights | openAccess |
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Except where otherwise noted, this item's license is described as: This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License 4.0 (CC BY-NC-ND 4.0), which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.