title: High-Throughput Lipidomic and Transcriptomic Analysis To Compare SP2/0, CHO, and HEK-293 Mammalian Cell Lines.
abstract: A combined lipidomics and transcriptomics analysis was performed on mouse myeloma SP2/0, Chinese hamster ovary (CHO), and human embryonic kidney (HEK) cells in order to compare widely used mammalian expression systems. Initial thin layer chromatography (TLC) analysis indicated that phosphatidylethanolamine (PE) and phosphatidylcholine (PC) were the major lipid components in all cell lines with lower amounts of sphingomyelin (SM) in SP2/0 compared to CHO and HEK, which was subsequently confirmed and expanded upon following mass spectrometry (MS) analysis. HEK contained 4-10-fold higher amounts of lyso phosphatidylethanolamine (LPE) and 2-4-fold higher amounts of lyso phosphatidylcholine (LPC) compared to SP2/0 and CHO cell lines. C18:1 followed by C16:1 were the main contributors to the difference in both LPE and LPC levels. Alternatively, the SP2/0 cell line exhibited 30-65-fold lower amounts of SM principally in the amount of 16:0. By mapping the transcriptomics data to KEGG pathways, we found expression levels of secretory phospholipase A2 (sPLA2), lysophospholipid acyltransferase (LPEAT), lysophosphatidylcholine acyltransferase (LPCAT), and lysophospholipase (LYPLA) can contribute to the differences in LPE and LPC. Sphingomyelin synthases (SMS) and sphingomyelin phosphodiesterase (SMase) enzymes may play roles in SM differences across the three cell lines. The results of this study provide insights that will aid the understanding of the physiological and secretory differences across recombinant protein production systems.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University , Baltimore, Maryland 21218, United States.
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Antibody Discovery and Protein Engineering, MedImmune LLC , Gaithersburg, Maryland 20878, United States.
authors.forename: Deniz
authors.initials: D
authors.lastname: Baycin-Hizal
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University , Baltimore, Maryland 21218, United States.
authors.forename: Amit
authors.identifier: 0000-0003-2140-5130
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University , Baltimore, Maryland 21218, United States.
authors.forename: Joseph
authors.initials: J
authors.lastname: Priola
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University , Baltimore, Maryland 21218, United States.
authors.forename: Michelle
authors.initials: M
authors.lastname: Bahri
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University , Baltimore, Maryland 21218, United States.
authors.forename: Kelley M
authors.initials: KM
authors.lastname: Heffner
authors.affiliation: Center for Metabolic Origins of Disease, Sanford Burnham Prebys Medical Discovery Institute , Orlando, Florida 32827, United States.
authors.forename: Miao
authors.initials: M
authors.lastname: Wang
authors.affiliation: Center for Metabolic Origins of Disease, Sanford Burnham Prebys Medical Discovery Institute , Orlando, Florida 32827, United States.
authors.forename: Xianlin
authors.initials: X
authors.lastname: Han
authors.affiliation: Antibody Discovery and Protein Engineering, MedImmune LLC , Gaithersburg, Maryland 20878, United States.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Bowen
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University , Baltimore, Maryland 21218, United States.
authors.forename: Michael J
authors.identifier: 0000-0002-1237-5550
authors.initials: MJ
authors.lastname: Betenbaugh
mesh_terms: D000818:Animals; D016466:CHO Cells; D002460:Cell Line; D002855:Chromatography, Thin Layer; D006224:Cricetinae; D003412:Cricetulus; D057809:HEK293 Cells; D006801:Humans; D008244:Lysophosphatidylcholines / Q000032:analysis*; D008246:Lysophospholipids / Q000032:analysis*; D013058:Mass Spectrometry; D051379:Mice; D010727:Phosphoric Diester Hydrolases / Q000235:genetics; D025341:Principal Component Analysis; D012333:RNA, Messenger / Q000378:metabolism; D013109:Sphingomyelins / Q000032:analysis*; D059467:Transcriptome; D017855:Transferases (Other Substituted Phosphate Groups) / Q000235:genetics
chemical_list: D008244:Lysophosphatidylcholines; D008246:Lysophospholipids; D012333:RNA, Messenger; D013109:Sphingomyelins; C008301:lysophosphatidylethanolamine; D017855:Transferases (Other Substituted Phosphate Groups); C019863:ceramide cholinephosphotransferase; D010727:Phosphoric Diester Hydrolases; C055160:sphingomyelin phosphodiesterase D