دورية أكاديمية
Comprehensive Evaluation of a Quantitative Shotgun Lipidomics Platform for Mammalian Sample Analysis on a High-Resolution Mass Spectrometer.
| العنوان: | Comprehensive Evaluation of a Quantitative Shotgun Lipidomics Platform for Mammalian Sample Analysis on a High-Resolution Mass Spectrometer. |
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| المؤلفون: | Nielsen IØ; Unit for Cell Death and Metabolism, Center for Autophagy, Recycling and Disease, Danish Cancer Society Research Center, Copenhagen DK-2100, Denmark., Vidas Olsen A; Computational Biology Laboratory, Center for Autophagy, Recycling and Disease, Danish Cancer Society Research Center, Copenhagen DK-2100, Denmark., Dicroce-Giacobini J; Unit for Cell Death and Metabolism, Center for Autophagy, Recycling and Disease, Danish Cancer Society Research Center, Copenhagen DK-2100, Denmark., Papaleo E; Computational Biology Laboratory, Center for Autophagy, Recycling and Disease, Danish Cancer Society Research Center, Copenhagen DK-2100, Denmark.; Translational Disease Systems Biology, Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen DK-2200, Denmark., Andersen KK; Unit for Statistics and Epidemiology, Danish Cancer Society Research Center, Copenhagen DK-2100, Denmark., Jäättelä M; Unit for Cell Death and Metabolism, Center for Autophagy, Recycling and Disease, Danish Cancer Society Research Center, Copenhagen DK-2100, Denmark.; Department of Cellular and Molecular Medicine, University of Copenhagen, Copenhagen DK-2200, Denmark., Maeda K; Unit for Cell Death and Metabolism, Center for Autophagy, Recycling and Disease, Danish Cancer Society Research Center, Copenhagen DK-2100, Denmark., Bilgin M; Unit for Cell Death and Metabolism, Center for Autophagy, Recycling and Disease, Danish Cancer Society Research Center, Copenhagen DK-2100, Denmark. |
| المصدر: | Journal of the American Society for Mass Spectrometry [J Am Soc Mass Spectrom] 2020 Apr 01; Vol. 31 (4), pp. 894-907. Date of Electronic Publication: 2020 Mar 04. |
| نوع المنشور: | Evaluation Study; Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: ACS Publications Country of Publication: United States NLM ID: 9010412 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1879-1123 (Electronic) Linking ISSN: 10440305 NLM ISO Abbreviation: J Am Soc Mass Spectrom Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: 2020- : Washington, DC : ACS Publications Original Publication: New York, NY : Elsevier, c1990- |
| مواضيع طبية MeSH: | Lipidomics/*instrumentation , Lipidomics/*methods , Lipids/*analysis , Mass Spectrometry/*instrumentation, Animals ; Brain Chemistry ; Cell Count ; Cell Culture Techniques ; Culture Media/chemistry ; Culture Media/pharmacology ; Female ; HeLa Cells ; High-Throughput Screening Assays ; Humans ; Limit of Detection ; Lipids/blood ; Lipids/chemistry ; MCF-7 Cells ; Mammals ; Mass Spectrometry/methods ; Mice ; NIH 3T3 Cells ; Reproducibility of Results ; Transfection |
| مستخلص: | Shotgun lipidomics is a powerful tool that enables simultaneous and fast quantification of diverse lipid classes through mass spectrometry based analyses of directly infused crude lipid extracts. We present here a shotgun lipidomics platform established to quantify 38 lipid classes belonging to four lipid categories present in mammalian samples and show the fine-tuning and comprehensive evaluation of its experimental parameters and performance. We first determined for all the targeted lipid classes the collision energy levels optimal for the recording of their lipid class- and species-specific fragment ions and fine-tuned the energy levels applied in the platform. We then performed a series of titrations to define the boundaries of linear signal response for the targeted lipid classes, and demonstrated that the dynamic quantification range spanned more than 3 orders of magnitude and reached sub picomole levels for 35 lipid classes. The platform identified 273, 261, and 287 lipid species in brain, plasma, and cultured fibroblast samples, respectively, at the respective optimal working sample amounts. The platform properly quantified the majority of these identified lipid species, while lipid species measured to be below the limit of quantification were efficiently removed from the data sets by the use of statistical analyses of data reproducibility or a cutoff threshold. Finally, we demonstrated that a series of parameters of cell culture conditions influence lipidomics outcomes, including confluency, medium supplements, and use of transfection reagents. The present study provides a guideline for setting up and using a simple and efficient platform for quantitatively exploring the mammalian lipidome. |
| فهرسة مساهمة: | Keywords: evaluation; high-resolution mass spectrometry; mammalian lipidome; shotgun lipidomics |
| المشرفين على المادة: | 0 (Culture Media) 0 (Lipids) |
| تواريخ الأحداث: | Date Created: 20200305 Date Completed: 20210222 Latest Revision: 20210222 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1021/jasms.9b00136 |
| PMID: | 32129994 |
| قاعدة البيانات: | MEDLINE |
Find this article in full text from Springer Verlag. 
Find this issue from the American Chemical Society | تدمد: | 1879-1123 |
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| DOI: | 10.1021/jasms.9b00136 |