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dc.contributor.authorde Faria E Silva, Ana Luiza
dc.contributor.authorElcoroaristizabal, Saioa
dc.contributor.authorRyder, Alan G.
dc.identifier.citationde Faria e Silva, Ana Luiza, Elcoroaristizabal, Saioa, & Ryder, Alan G. (2020). Multi-attribute quality screening of immunoglobulin G using polarized Excitation Emission Matrix spectroscopy. Analytica Chimica Acta, 1101, 99-110. doi:
dc.description.abstractImmunoglobulin G (IgG) is often used as a starting material for the production of functionalised antibodies, like Antibody Drug Conjugates (ADCs), PEGlyated-conjugates, or radioimmunoconjugates. The gross structural quality of the protein starting material is, therefore, an important factor in determining final product composition, purity, and quality. In terms of structural quality, one needs to know both the aggregation content and the tertiary structure of the protein. The measurement of structural quality in solution can thus be difficult, but the use of intrinsic fluorescence measurements might offer a solution because of its high sensitivity, ease of use, and when implemented in via multi-dimensional techniques like polarized Excitation Emission Matrix (pEEM) spectroscopy, its high information content. Here we demonstrate how pEEM measurements can be used as a multi-attribute screening method for protein quality using a polyclonal rabbit immunoglobulin (rIgG) model system. By using both Rayleigh scatter and fluorescence emission in combination with simple chemometric data analysis methods like Principal Component analysis (PCA) and unfolded partial least squares (U-PLS) one can simultaneously measure protein concentration, structural variance, and particle/aggregate composition. Furthermore, one can generate quantitative prediction models for non-reversible aggregation content as described by size exclusion chromatography (SEC) and obtain qualitative information about reversible aggregate content, which cannot be obtained from SEC measurements. In conclusion, the pEEM measurement approach is a potentially useful Process Analytical Technology (PAT) method for downstream processing operations in biopharmaceutical manufacturing.en_IE
dc.description.sponsorshipThis publication has emanated from research supported in part by a research grant from Science Foundation Ireland (SFI) and is co-funded under the European Regional Development Fund under Grand number (14/IA/2282, Advanced Analytics for Biological Therapeutic Manufacture, to AGR). We also thank Agilent Technologies (Mulgrave Victoria, Australia) for the loan of a fluorescence spectrometer.en_IE
dc.relation.ispartofAnalytica Chimica Actaen
dc.subjectImmunoglobulin Gen_IE
dc.subjectAggregation. stdeven_IE
dc.titleMulti-attribute quality screening of immunoglobulin G using polarized Excitation Emission Matrix spectroscopyen_IE
dc.contributor.funderScience Foundation Irelanden_IE
dc.contributor.funderEuropean Regional Development Funden_IE
dc.local.contactAlan Ryder, School Of Chemistry, Room 213, Arts/Science Building, South Cam, Nui Galway. 2943 Email:
dcterms.projectinfo:eu-repo/grantAgreement/SFI/SFI Investigator Programme/14/IA/2282/IE/Advanced Analytics for Biological Therapeutic Manufacture (AA-BTM)./en_IE

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