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On-line sequential injection-capillary electrophoresis for near-real-time monitoring of extracellular lactate in cell culture flasks
journal contribution
posted on 2014-01-03, 00:00 authored by A A Alhusban, A J Gaudry, M C Breadmore, N Gueven, Rosanne GuijtRosanne GuijtCell culture has replaced many in vivo studies because of ethical and regulatory measures as well as the possibility of increased throughput. Analytical assays to determine (bio)chemical changes are often based
on end-point measurements rather than on a series of sequential determinations. The purpose of this work is to develop an analytical system for monitoring cell culture based on sequential injection-capillary
electrophoresis (SI-CE) with capacitively coupled contactless conductivity detection (C4D). The system was applied for monitoring lactate production, an important metabolic indicator, during mammalian cell
culture. Using a background electrolyte consisting of 25 mM tris(hydroxymethyl)aminomethane, 35 mM
cyclohexyl-2-aminoethanesulfonic acid with 0.02% poly(ethyleneimine)(PEI) at pH 8.65 and a multilayer polymer coated capillary, lactate could be resolved from other compounds present in media with relative standard deviations 0.07% for intraday electrophoretic mobility and an analysis time of less than 10 min. Using the human embryonic kidney cell line HEK293, lactate concentrations in the cell culture medium were measured every 20 min over 3 days, requiring only 8.73 L of sample per run. Combining simplicity, portability, automation, high sample throughput, low limits of detection, low sample consumption and the ability to up- and outscale,this new methodology represents a promising technique for near real-time monitoring of chemical changes in diverse cell culture applications.
on end-point measurements rather than on a series of sequential determinations. The purpose of this work is to develop an analytical system for monitoring cell culture based on sequential injection-capillary
electrophoresis (SI-CE) with capacitively coupled contactless conductivity detection (C4D). The system was applied for monitoring lactate production, an important metabolic indicator, during mammalian cell
culture. Using a background electrolyte consisting of 25 mM tris(hydroxymethyl)aminomethane, 35 mM
cyclohexyl-2-aminoethanesulfonic acid with 0.02% poly(ethyleneimine)(PEI) at pH 8.65 and a multilayer polymer coated capillary, lactate could be resolved from other compounds present in media with relative standard deviations 0.07% for intraday electrophoretic mobility and an analysis time of less than 10 min. Using the human embryonic kidney cell line HEK293, lactate concentrations in the cell culture medium were measured every 20 min over 3 days, requiring only 8.73 L of sample per run. Combining simplicity, portability, automation, high sample throughput, low limits of detection, low sample consumption and the ability to up- and outscale,this new methodology represents a promising technique for near real-time monitoring of chemical changes in diverse cell culture applications.
History
Journal
Journal of chromatography AVolume
1323Pagination
157 - 162Publisher
ElsevierLocation
Amsterdam, The NetherlandsPublisher DOI
ISSN
0021-9673eISSN
1873-3778Language
engPublication classification
C Journal article; C1.1 Refereed article in a scholarly journalCopyright notice
2013, ElsevierUsage metrics
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No categories selectedKeywords
sequential injection capillary electrophoresisonline monitoringbio process monitoringcapillary electrophoresiscell cultureScience & TechnologyLife Sciences & BiomedicinePhysical SciencesBiochemical Research MethodsChemistry, AnalyticalBiochemistry & Molecular BiologyChemistrySequential injection capillaryelectrophoresisELECTROKINETIC INJECTIONLACTIC-ACIDISOTACHOPHORESISFERMENTATIONMETABOLISM
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