Matching IR-MALDI-o-TOF Mass Spectrometry with the TLC Overlay Binding Assay and Its Clinical Application for Tracing Tumor-Associated Glycosphingolipids in Hepatocellular and Pancreatic Cancer
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- 作者：Ute Distler ; Marcel Hü ; lsewig ; Jamal Souady ; Klaus Dreisewerd ; Jö ; rg Haier ; Norbert Senninger ; Alexander W. Friedrich ; Helge Karch ; Franz Hillenkamp ; Stefan Berkenkamp ; Jasna Peter-Katalini ; Jo
- 刊名：Analytical Chemistry
- 出版年：2008
- 出版时间：March 15, 2008
- 年：2008
- 卷：80
- 期：6
- 页码：1835 - 1846
- 全文大小：622K
- 年卷期：v.80,no.6(March 15, 2008)
- ISSN：1520-6882
文摘
Glycosphingolipids (GSLs), composed of a hydrophiliccarbohydrate chain and a lipophilic ceramide anchor, playpivotal roles in countless biological processes, includingthe development of cancer. As part of the investigation ofthe vertebrate glycome, GSL analysis is undergoing rapidexpansion owing to the application of modern massspectrometry. Here we introduce direct coupling of IR-MALDI-o-TOF mass spectrometry with the TLC overlaybinding assay for the structural characterization of GSLs.We matched three complementary methods including (i)TLC separation of GSLs, (ii) their detection with oligosaccharide-specific proteins, and (iii) in situ MS analysis ofprotein-detected GSLs. The high specificity and sensitivityis demonstrated by use of antibodies, bacterial toxins, anda plant lectin. The procedure works on a nanogram scale,and detection limits of less than 1 ng at its best ofimmunostained GSLs were obtained. Furthermore, onlycrude lipid extracts of biological sources are required forTLC-IR-MALDI-MS, omitting any laborious GSL downstream purification procedures. This strategy was successfully applied to the identification of cancer-associatedGSLs in human hepatocellular and pancreatic tumors.Thus, the in situ TLC-IR-MALDI-MS of immunolabeledGSLs opens new doors by delivering specific structuralinformation of trace quantities of GSLs with only a limitedinvestment in sample preparation.