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4D-脂质组学高通量工作流程
2021/03/05 15:47
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Lipids represent an important compound class in biological systems as they play key roles in cell membranes, in the energy storage or as signaling molecules.
This makes them interesting markers for studying biological systems regarding clinical research. At the same time, lipids depict a highly complex class of compounds. Both, the
structural diversity as well as the high dynamic range in concentrations makes the analysis of lipid extracts challenging. A major bottleneck in lipid profiling is the annotation and subsequent validation of lipid assignments. Additionally, a high throughput is often necessary, e.g. in phenomics related large cohort studies.
Using the timsTOF Pro or timsTOF fleX systems, a high MS/MS coverage can be achieved by the PASEF® acquisition mode, even with single injections and when using fast LC gradients. Furthermore, the trapped ion mobility separation cleans up the MS/MS spectra and improves the matching of fragmentation rules or spectral libraries. As a third factor of improvement, the
automatically acquired CCS values can be directly used to increase the confidence in annotations. This 4D-Lipidomics workflow is demonstrated in detail in the following.
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PASEF应用于COVID-19的高通量定量大规模血浆蛋白质组学研究
However, the dynamic range of protein concentration in these samples poses a challenge for in-depth proteome quantification. PASEF® is a powerful technology for rapid in-depth and sensitive proteome quantification that is also applicable to biofluids like plasma and serum. We combined PASEF with high-throughput gradient settings for the analysis of hundreds of plasma and serum samples and quantified several hundred protein groups routinely in 21- and 47-minute runtimes. The timsTOF Pro coupled with the Evosep One nanoflow UPLC delivers quantitation with median CV less than 20% and quantified more than 70 known biomarkers in a study investigating over 700 COVID-19 serum samples
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MALDI Guided SpatialOMx，即基于质谱成像定位的空间多组学分析，完美的将MALDI成像技术和传统LC-MS/MS组学流程结合在了一起，它的特点是利用了MALDI成像技术能在分子原位对生物分子进行定位的特点，在生物组织的内部锁定目标微区（ROI，Region of Interests），在对该目标微区实施激光微切割（LCM，laser capture microdissection）后，进行LC-MS/MS组学分析，从而实现了深度的蛋白组信息挖掘。
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最新一代布鲁克timsTOF fleX使用创新的激光后电离MALDI-2技术获得了较传统MALDI 高1-3个数量级的灵敏度。MALDI-2提高了离子产率并降低了离子抑制效应。同时，MALDI-2还可对传统MALDI不能电离的药物和代谢物分子检测和成像，如甾醇、他汀类、激素、维生素、聚糖等生物分子。这种实质性的改进对于DMPK研究尤其重要。
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