Tokuoka Suzumi M, Kita Yoshihiro, Sato Masaya, Shimizu Takao, Yatomi Yutaka, Oda Yoshiya
Department of Lipidomics, Graduate School of Medicine, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-8654, Japan.
Department of Clinical Laboratory Medicine, Graduate School of Medicine, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo 113-8654, Japan.
Metabolites. 2020 Dec 30;11(1):19. doi: 10.3390/metabo11010019.
In clinical lipidomics, it is a challenge to measure a large number of samples and to reproduce the quantitative results. We expanded the range of application of the tandem mass tag (TMT) method, which is widely used in proteomics, to lipidomic fields. There are various types of lipid molecule, for example, eicosanoids have a carboxyl group and phosphatidic acid has a phosphate group. We modified these functional groups simultaneously with TMT. This approach allows for a single analysis by mixing six samples and using one of the six samples as a bridging sample; the quantitative data can be easily normalized even if the number of measurements increases. To accommodate a large number of samples, we utilize a pooled serum sample of 300 individuals as a bridging sample. The stability of these lipid molecules in serum was examined as an analytical validation for the simultaneous TMT labeling. It was found that the stability of these lipid molecules in serum differs greatly depending on the lipid species. These findings reaffirmed the importance of proper sample preparation and storage to obtain reliable data. The TMT labeling method is expected to be a useful method for lipidomics with high-throughput and reliable reproducibility.
在临床脂质组学中，对大量样本进行测量并重现定量结果是一项挑战。我们将广泛应用于蛋白质组学的串联质量标签（TMT）方法的应用范围扩展到了脂质组学领域。脂质分子有多种类型，例如，类二十烷酸有一个羧基，磷脂酸有一个磷酸基团。我们用TMT同时修饰这些官能团。这种方法允许通过混合六个样本并将六个样本中的一个用作桥接样本进行单次分析；即使测量次数增加，定量数据也能轻松归一化。为了处理大量样本，我们使用300名个体的混合血清样本作为桥接样本。作为对同时进行TMT标记的分析验证，我们检测了这些脂质分子在血清中的稳定性。结果发现，这些脂质分子在血清中的稳定性因脂质种类不同而有很大差异。这些发现再次证实了进行适当的样本制备和储存以获得可靠数据的重要性。TMT标记方法有望成为一种用于脂质组学的具有高通量和可靠重现性的有用方法。
