Lectin affinity capillary electrophoresis in glycoform analysis applying the partial filling technique
Lectin affinity capillary electrophoresis in glycoform analysis applying the partial filling technique
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DOI：
10.1016/j.jchromb.2004.06.042
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发表时间：
2004-10-05
影响因子：
3
通讯作者：
Ohlson, S
中科院分区：
文献类型：
--
作者：
Bergström, M;Nilsson, M;Ohlson, S
蛋白质糖基化及其在生物相互作用中的意义的研究日益受到人们的关注。这项工作展示了一种基于凝集素的α(1)-酸性糖蛋白(AGP或类粘蛋白)的蛋白质糖型的毛细管电泳法。以凝集素刀豆蛋白A(ConA)为亲和配基，采用“部分填充技术”进行糖型分析。ConA将人AGP分成两个峰；第一个峰包括不含双天线多糖的AGP糖型，第二个峰代表含有一个或多个双天线多糖的部分。通过对临床样品中AGP和N-糖苷酶F处理的AGP的分析，证明了该方法的适用性。AGP分离也被用作报告系统来估算ConA与竞争糖之间的解离常数(K-D)。(C)2004爱思唯尔B.V.保留所有权利。
The study of protein glycosylation and its significance in biological interactions is a field of growing interest. This work demonstrates a lectin-based separation of protein glycoforms of alpha(1)-acid glycoprotein (AGP or orosomucoid) with capillary electrophoresis. Glycoform analysis was performed with a "partial filling technique" with the lectin Concanavalin A (Con A) as affinity ligand. Con A separated human AGP into two peaks; the first peak included AGP glycoforms without biantennary glycans, and the second peak represented the fraction that had one or more biantennary glycans. The applicability of the method was demonstrated with the analysis of AGP from clinical samples and AGP treated with N-glycosidase F. The AGP separation was also used as a reporter system to estimate the dissociation constant (K-D) between Con A and a competing sugar. (C) 2004 Elsevier B.V. All rights reserved.