Physical characterixation and in vitro biological impact of highly aggregated antibodies separated into size-enriched populations by fluorescence-activated cell sorting
dc.contributor.author | Telikepalli, Srivalli | |
dc.contributor.author | Shinogle, Heather E. | |
dc.contributor.author | Thapa, Prem S. | |
dc.contributor.author | Kim, Jae Hyun | |
dc.contributor.author | Meghana, Deshpande | |
dc.contributor.author | Jawa, Vibha | |
dc.contributor.author | Middaugh, C. Russell | |
dc.contributor.author | Narhi, Linda O. | |
dc.contributor.author | Joubert, Marisa K. | |
dc.contributor.author | Volkin, David B. | |
dc.date.accessioned | 2017-04-26T16:33:37Z | |
dc.date.available | 2017-04-26T16:33:37Z | |
dc.date.issued | 2015-05 | |
dc.identifier.citation | Telikepalli, S., Shinogle, H. E., Thapa, P. S., Kim, J. H., Deshpande, M., Jawa, V., … Volkin, D. B. (2015). Physical characterization and in vitro biological impact of highly aggregated antibodies separated into size-enriched populations by fluorescence-activated cell sorting. Journal of Pharmaceutical Sciences, 104(5), 1575–1591. http://doi.org/10.1002/jps.24379 | en_US |
dc.identifier.uri | http://hdl.handle.net/1808/23809 | |
dc.description.abstract | An IgG2 monoclonal antibody (mAb) solution was subjected to stirring, generating high concentrations of nanometer and subvisible particles, which were then successfully size enriched into different size bins by low speed centrifugation or a combination of gravitational sedimentation and Fluorescence-Activated Cell Sorting (FACS). The size-fractionated mAb particles were assessed for their ability to elicit the release of cytokines from a population of donor-derived human peripheral blood mononuclear cells (PBMC) at two phases of the immune response. Fractions enriched in nanometer-sized particles showed a lower response than those enriched in micron-sized particles in this assay. Particles of 5–10 μm in size displayed elevated cytokine release profiles compared to other size ranges. Stir-stressed mAb particles had amorphous morphology, contained protein with partially altered secondary structure, elevated surface hydrophobicity (compared to controls), and trace levels of elemental fluorine. FACS size-enriched the mAb particle samples, yet did not notably alter the overall morphology or composition of particles as measured by Microflow imaging, Transmission Electron Microscopy, and Scanning Electron Microscopy-Energy Dispersive X-ray Spectroscopy. The utility and limitations of FACS for size separation of mAb particles and potential of in-vitro PBMC studies to rank order the immunogenic potential of various types of mAb particles is discussed. | 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 | Proteins | en_US |
dc.subject | Protein aggregation | en_US |
dc.subject | Particles | en_US |
dc.subject | Monoclonal antibody | en_US |
dc.subject | IgG | en_US |
dc.subject | Immune response | en_US |
dc.subject | Immunogenicity | en_US |
dc.subject | PBMC | en_US |
dc.subject | In-vitro | en_US |
dc.title | Physical characterixation and in vitro biological impact of highly aggregated antibodies separated into size-enriched populations by fluorescence-activated cell sorting | en_US |
dc.type | Article | en_US |
kusw.kuauthor | Telikepalli, Srivalli | |
kusw.kuauthor | Kim, Jae Hyun | |
kusw.kuauthor | Middaugh, C. Russell | |
kusw.kuauthor | Volkin, David B. | |
kusw.kudepartment | Pharmaceutical Chemistry | en_US |
dc.identifier.doi | 10.1002/jps.24379 | 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 | PMC4448733 | 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.