{"doi":"10.1002/elps.201400451","title":"Quantification of riboflavin, flavin mononucleotide, and flavin adenine dinucleotide in mammalian model cells by CE with LED‐induced fluorescence detection","abstract":"<jats:p>Cultured mammalian cells essential are model systems in basic biology research, production platforms of proteins for medical use, and testbeds in synthetic biology. Flavin cofactors, in particular flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), are critical for cellular redox reactions and sense light in naturally occurring photoreceptors and optogenetic tools. Here, we quantified flavin contents of commonly used mammalian cell lines. We first compared three procedures for extraction of free and noncovalently protein‐bound flavins and verified extraction using fluorescence spectroscopy. For separation, two CE methods with different BGEs were established, and detection was performed by LED‐induced fluorescence with limit of detections (LODs 0.5–3.8 nM). We found that riboflavin (RF), FMN, and FAD contents varied significantly between cell lines. RF (3.1–14 amol/cell) and FAD (2.2–17.0 amol/cell) were the predominant flavins, while FMN (0.46–3.4 amol/cell) was found at markedly lower levels. Observed flavin contents agree with those previously extracted from mammalian tissues, yet reduced forms of RF were detected that were not described previously. Quantification of flavins in mammalian cell lines will allow a better understanding of cellular redox reactions and optogenetic tools.</jats:p>","journal":"ELECTROPHORESIS","year":2015,"id":661020,"datarank":0.6307038929086449,"base_score":4.204692619390966,"endowment":4.204692619390966,"self_citation_contribution":0.6307038929086449,"citation_network_contribution":0.0,"self_endowment_contribution":0.6307038929086449,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":66,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1725576,"name":"Álvaro Ingles‐Prieto","orcid":null,"position":1,"is_corresponding":false},{"id":309066,"name":"Christian Neusüß","orcid":"0000-0003-2404-4924","position":2,"is_corresponding":false},{"id":1188173,"name":"Michael Lämmerhofer","orcid":null,"position":3,"is_corresponding":false},{"id":583332,"name":"Harald Janovjak","orcid":"0000-0002-8023-9315","position":4,"is_corresponding":false},{"id":1725574,"name":"Jens Hühner","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Quantification of riboflavin, flavin mononucleotide, and flavin adenine dinucleotide in mammalian model cells by CE with LED‐induced fluorescence detection","abstract":"<jats:p>Cultured mammalian cells essential are model systems in basic biology research, production platforms of proteins for medical use, and testbeds in synthetic biology. Flavin cofactors, in particular flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), are critical for cellular redox reactions and sense light in naturally occurring photoreceptors and optogenetic tools. Here, we quantified flavin contents of commonly used mammalian cell lines. We first compared three procedures for extraction of free and noncovalently protein‐bound flavins and verified extraction using fluorescence spectroscopy. For separation, two CE methods with different BGEs were established, and detection was performed by LED‐induced fluorescence with limit of detections (LODs 0.5–3.8 nM). We found that riboflavin (RF), FMN, and FAD contents varied significantly between cell lines. RF (3.1–14 amol/cell) and FAD (2.2–17.0 amol/cell) were the predominant flavins, while FMN (0.46–3.4 amol/cell) was found at markedly lower levels. Observed flavin contents agree with those previously extracted from mammalian tissues, yet reduced forms of RF were detected that were not described previously. Quantification of flavins in mammalian cell lines will allow a better understanding of cellular redox reactions and optogenetic tools.</jats:p>","is_dataset_classified":null,"base_score":4.204692619390966,"endowment":4.204692619390966,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"25488801","pmcid":null,"openalex_id":"https://openalex.org/W2081785349","authors":[],"funders":[{"funder_name":"Human Frontier Science Program","grant_id":"RGY0084_2012","title":null},{"funder_name":"Deutsche Forschungsgemeinschaft","grant_id":"INST 37/821-1 FUGG","title":null},{"funder_name":"Dan David Prize","grant_id":"Scholarship","title":null},{"funder_name":"European Union 7th Framework Programme","grant_id":"CIG-303564","title":null},{"funder_name":"Deutsche Forschungsgemeinschaft","grant_id":"unidentified","title":"unidentified"},{"funder_name":"Ramon Areces Foundation","grant_id":"","title":null},{"funder_name":"Struktur- und Innovationsfonds Baden-Württemberg","grant_id":"","title":null}],"total_grants":7,"fwci":1.4322,"citation_percentile":0.81019121,"influential_citations":0,"citation_trend":[{"year":2016,"count":7},{"year":2017,"count":4},{"year":2018,"count":6},{"year":2019,"count":9},{"year":2020,"count":4},{"year":2021,"count":4},{"year":2022,"count":8},{"year":2023,"count":6},{"year":2024,"count":6},{"year":2025,"count":6},{"year":2026,"count":6}],"oa_status":"closed","license":"Wiley Online Library User Agreement","oa_locations":[{"url":"https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Felps.201400451","host_type":"publisher"},{"url":"https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/pdf/10.1002/elps.201400451","host_type":"publisher"},{"url":"https://doi.org/10.1002/elps.201400451","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/25488801","host_type":"repository"},{"url":"https://dx.doi.org/10.1002/elps.201400451","host_type":""},{"url":"https://doi.org/https://doi.org/10.1002/elps.201400451","host_type":""}],"fields_of_study":["Advanced biosensing and bioanalysis techniques","Photoreceptor and optogenetics research","Biosensors and Analytical Detection","0301 basic medicine","0303 health sciences","03 medical and health sciences","Animals","CHO Cells","Calibration","Cell Line","Cells, Cultured","Cricetulus","Electrophoresis, Capillary","Flavin Mononucleotide","Flavin-Adenine Dinucleotide","HEK293 Cells","Humans","Lasers, Semiconductor","Mammals","Mice","NIH 3T3 Cells","Reproducibility of Results","Riboflavin","Spectrometry, Fluorescence"],"mesh_terms":["Animals","Calibration","Cell Line","Cells, Cultured","Cricetulus","Flavin-Adenine Dinucleotide","Flavin Mononucleotide","Humans","Mammals","Riboflavin","Spectrometry, Fluorescence","Reproducibility of Results","CHO Cells","Electrophoresis, Capillary","NIH 3T3 Cells","Mice","Lasers, Semiconductor","HEK293 Cells"],"keywords":["Flavin mononucleotide","Riboflavin","Flavin adenine dinucleotide","Flavin group","Flavoprotein","Fluorescence","Chemistry","Biochemistry","Enzyme","Cofactor","Capillary electrophoresis","Fmn","Fad","Optogenetics","Light-emitting Diode-induced Fluorescence Detection","Mammals","570","Electrophoresis, Capillary","Reproducibility of Results","CHO Cells","540","Cell Line","Mice","Cricetulus","HEK293 Cells","Spectrometry, Fluorescence","Calibration","Flavin-Adenine Dinucleotide","NIH 3T3 Cells","Animals","Humans","Lasers, Semiconductor","Cells, Cultured"],"sdg_mappings":[{"sdg_number":3,"sdg_label":"3. 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