Madera Milan, Mechref Yehia, Klouckova Iveta, Novotny Milos V
Department of Chemistry, Indiana University, Bloomington, Indiana 47405, USA.
J Proteome Res. 2006 Sep;5(9):2348-63. doi: 10.1021/pr060169x.
We describe an effective analytical approach to identify trace glycoproteins in a small volume of human serum. The system is based on automatable affinity enrichment through silica-based lectin microcolumns and a further separation of the retained glycoproteins on a reversed-phase liquid chromatography with superficially porous packing, operating at high temperature. The fractionated sample is further directed into a 96-well plate for trypsinization and LC-MS/MS analysis. Using a major-component depleted blood serum (16 microg total protein), we were able to identify 271 glycoproteins through this analytical system.
我们描述了一种有效的分析方法，用于在少量人血清中鉴定痕量糖蛋白。该系统基于通过硅胶基凝集素微柱进行的可自动化亲和富集，以及在高温下运行的具有表面多孔填料的反相液相色谱上对保留的糖蛋白进行进一步分离。分馏后的样品被进一步导入 96 孔板中进行胰蛋白酶消化和 LC-MS/MS 分析。使用主要成分耗尽的血清（总蛋白 16 微克），我们通过该分析系统能够鉴定出 271 种糖蛋白。
