dc.contributor.advisor | Schöneich, Christian | |
dc.contributor.author | Yang, Yi | |
dc.date.accessioned | 2016-10-11T18:56:12Z | |
dc.date.available | 2016-10-11T18:56:12Z | |
dc.date.issued | 2016-05-31 | |
dc.date.submitted | 2016 | |
dc.identifier.other | http://dissertations.umi.com/ku:14473 | |
dc.identifier.uri | http://hdl.handle.net/1808/21654 | |
dc.description.abstract | In the biotechnology industry, oxidative carbonylation as a post-translational modification of protein pharmaceuticals has not been studied in detail. Using Quality by Design (QbD) principles, understanding the impact of oxidative carbonylation on product quality of protein pharmaceuticals, particularly from a site-specific perspective, is critical. However, comprehensive identification of carbonylation sites has so far remained a very difficult analytical challenge for the industry. In this work, for the first time the identification of specific carbonylation sites on recombinant monoclonal antibodies was reported using a new analytical approach via derivatization with Girard's Reagent T (GRT) and subsequent peptide mapping with high-resolution mass spectrometry. Enhanced ionization efficiency and high quality MS2 data resulted from GRT derivatization were observed as key benefits of this approach, which enabled direct identification of carbonylation sites without any fractionation or affinity enrichment steps. A simple data filtering process was also incorporated to significantly reduce false positive assignments. Sensitivity and efficiency of this approach were demonstrated by identification of carbonylation sites on both unstressed and oxidized antibody bulk drug substances. The applicability of this approach was further demonstrated by identification of 14 common carbonylation sites on three highly similar IgG1s. This approach represents a significant improvement to the existing analytical methodologies and facilitates extended characterization of oxidative carbonylation on recombinant monoclonal antibodies and potentially other protein pharmaceuticals in the biotechnology industry. | |
dc.format.extent | 60 pages | |
dc.language.iso | en | |
dc.publisher | University of Kansas | |
dc.rights | Copyright held by the author. | |
dc.subject | Pharmaceutical sciences | |
dc.subject | critical quality attributes | |
dc.subject | metal catalyzed oxidation | |
dc.subject | oxidative carbonylation | |
dc.subject | post-translational modification | |
dc.subject | recombinant monoclonal antibody | |
dc.title | Site-Specific Characterization of Oxidative Carbonylation on Recombinant Monoclonal Antibodies | |
dc.type | Thesis | |
dc.contributor.cmtemember | Stobaugh, John F. | |
dc.contributor.cmtemember | Gennaro, Lynn | |
dc.thesis.degreeDiscipline | Pharmaceutical Chemistry | |
dc.thesis.degreeLevel | M.S. | |
dc.identifier.orcid | | |
dc.identifier.orcid | https://orcid.org/0000-0002-7869-2858 | |
dc.provenance | 04/05/2017: The ETD release form is attached to this record as a license file. | |
dc.rights.accessrights | openAccess | |