Sparbier Katrin, Wenzel Thomas, Kostrzewa Markus
Bruker Daltonik GmbH, Permoserstrasse 15, 04318 Leipzig, Germany.
J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Aug 7;840(1):29-36. doi: 10.1016/j.jchromb.2006.06.028. Epub 2006 Jul 12.
Concanavalin A, boronic acid and Wheat germ agglutinin functionalized magnetic micro-particles were developed to enrich glycosylated peptides and proteins. The bead functionalities were validated according to their specificity by analyses of model proteins. Validated beads were employed for the enrichment of glycosylated human serum proteins. Eluted glycoproteins were digested by trypsin and the resulting peptides were purified by magnetic MB-HIC C8 beads. Each fraction was analyzed by MALDI-TOF MS and single peaks were subjected to MALDI-TOF/TOF MS with the objective to identify the respective proteins by database search. Search results revealed overlapping profiles of known serum glycoproteins.
开发了伴刀豆球蛋白A、硼酸和麦胚凝集素功能化的磁性微粒，用于富集糖基化肽和蛋白质。通过对模型蛋白的分析，根据其特异性验证了磁珠的功能。经验证的磁珠用于富集糖基化的人血清蛋白。洗脱的糖蛋白用胰蛋白酶消化，所得肽段用磁性MB-HIC C8磁珠纯化。对每个馏分进行基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）分析，对单峰进行基质辅助激光解吸电离串联飞行时间质谱（MALDI-TOF/TOF MS）分析，目的是通过数据库搜索鉴定相应的蛋白质。搜索结果显示了已知血清糖蛋白的重叠图谱。
