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dc.contributor.authorGroza, RC,Calvet, A,Ryder, AG
dc.contributor.authorGroza, Radu Constantin
dc.contributor.authorCalvet, Amandine
dc.contributor.authorRyder, Alan G.
dc.date.accessioned2014-09-09T12:29:22Z
dc.date.available2014-09-09T12:29:22Z
dc.date.issued2014-03-12
dc.identifier.citationA Fluorescence Anisotropy Method for Protein Concentration Monitoring in Complex Cell Culture Media. R.C. Groza, A. Calvet, and A.G. Ryder. Analytica Chimica Acta, 821, 54-61, (2014).en_US
dc.identifier.urihttp://hdl.handle.net/10379/4521
dc.descriptionJournal articleen_US
dc.description.abstractThe rapid, quantitative analysis of the complex cell culture media used in biopharmaceutical manufacturing is of critical importance. Requirements for cell culture media composition profiling, or changes in specific analyte concentrations (e. g. amino acids in the media or product protein in the bioprocess broth) often necessitate the use of complicated analytical methods and extensive sample handling. Rapid spectroscopic methods like multi-dimensional fluorescence (MDF) spectroscopy have been successfully applied for the routine determination of compositional changes in cell culture media and bioprocess broths. Quantifying macromolecules in cell culture media is a specific challenge as there is a need to implement measurements rapidly on the prepared media. However, the use of standard fluorescence spectroscopy is complicated by the emission overlap from many media components. Here, we demonstrate how combining anisotropy measurements with standard total synchronous fluorescence spectroscopy (TSFS) provides a rapid, accurate quantitation method for cell culture media. Anisotropy provides emission resolution between large and small fluorophores while TSFS provides a robust measurement space. Model cell culture mediawas prepared using yeastolate (2.5mgmL(-1)) spiked with bovine serum albumin (0 to 5mgmL (-1)). Using this method, protein emission is clearly discriminated from background yeastolate emission, allowing for accurate bovine serum albumin (BSA) quantification over a 0.1 to 4.0mgmL(-1)range with a limit of detection (LOD) of 13.8mgmL(-1). (c) 2014 Published by Elsevier B.V.en_US
dc.description.sponsorshipIrish Research Council - EMBARK Initiativeen_US
dc.formatapplication/MSWorden_US
dc.language.isoenen_US
dc.publisherElsevieren_US
dc.relation.ispartofAnalytica chimica actaen
dc.subjectFluorescenceen_US
dc.subjectAnisotropyen_US
dc.subjectTotal synchronous fluorescence spectroscopy (TSFS)en_US
dc.subjectBioprocessen_US
dc.subjectProteinen_US
dc.titleA fluorescence anisotropy method for measuring protein concentration in complex cell culture mediaen_US
dc.typeArticleen_US
dc.date.updated2014-09-07T10:48:30Z
dc.identifier.doi10.1016/j.aca.2014.03.007
dc.local.publishedsourcehttp://dx.doi.org/10.1016/j.aca.2014.03.007en_US
dc.description.peer-reviewedpeer-reviewed
dc.contributor.funder|~|
dc.internal.rssid6299570
dc.local.contactAlan Ryder, School Of Chemistry, Room 228, Arts/Science Building, Nui Galway. 2943 Email: alan.ryder@nuigalway.ie
dc.local.copyrightcheckedYes
dc.local.versionACCEPTED
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