组织切片糖胺聚糖分析的敏感方法。
Sensitive method for glycosaminoglycan analysis of tissue sections.
机构信息
MS Proteomics Research Group, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Magyar Tudósok Körútja 2, H-1117, Budapest, Hungary.
MS Proteomics Research Group, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Magyar Tudósok Körútja 2, H-1117, Budapest, Hungary.
出版信息
J Chromatogr A. 2018 Apr 6;1544:41-48. doi: 10.1016/j.chroma.2018.02.034. Epub 2018 Feb 25.
Abstract
A simple, isocratic HPLC method based on HILIC-WAX separation, has been developed for analyzing sulfated disaccharides of glycosaminoglycans (GAGs). To our best knowledge, this is the first successful attempt using this special phase in nano-HPLC-MS analysis. Mass spectrometry was based on negative ionization, improving both sensitivity and specificity. Detection limit for most sulfated disaccharides were approximately 1 fmol; quantitation limits 10 fmol. The method was applied for glycosaminoglycan profiling of tissue samples, using surface digestion protocols. This novel combination provides sufficient sensitivity for GAG disaccharide analysis, which was first performed using prostate cancer tissue microarrays. Preliminary results show that GAG analysis may be useful for identifying cancer related changes in small amounts of tissue samples (ca. 10 μg).
摘要
建立了一种基于 HILIC-WAX 分离的简单等度 HPLC 方法，用于分析糖胺聚糖（GAG）的硫酸化二糖。据我们所知，这是首次在纳升 HPLC-MS 分析中成功使用该特殊固定相。质谱基于负离子化，提高了灵敏度和特异性。大多数硫酸化二糖的检测限约为 1 fmol；定量限为 10 fmol。该方法应用于组织样品的糖胺聚糖分析，使用表面消化方案。这种新的组合为 GAG 二糖分析提供了足够的灵敏度，这是首次在前列腺癌组织微阵列上进行的。初步结果表明，GAG 分析可能有助于鉴定小量组织样品（约 10 µg）中的癌症相关变化。
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