Analysis of carbohydrate‐mediated heterogeneity and characterization of N‐linked oligosaccharides of glycoproteins by high performance capillary electrophoresis
Analysis of carbohydrate‐mediated heterogeneity and characterization of N‐linked oligosaccharides of glycoproteins by high performance capillary electrophoresis
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通过高性能毛细管电泳分析碳水化合物介导的异质性和糖蛋白 N 连接寡糖的表征
DOI：
10.1002/elps.1150130174
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发表时间：
1992
期刊：
影响因子：
2.9
通讯作者：
D. Ferrier
中科院分区：
文献类型：
--
作者：
M. Taverna;A. Baillet;D. Biou;M. Schlüter;R. Werner;D. Ferrier
毛细管区带电泳（CZE）已被研究作为一种替代方法来分析糖蛋白的糖部分。检查了重组纤溶酶原激活剂（rt-PA）的碳水化合物介导的微观异质性。使用CZE将糖蛋白解析为多个电泳物种，但由于分子吸附到毛细管壁而使分离复杂化。考察了pH值、缓冲液浓度和阳离子添加剂等因素对糖肽分离的影响。使用100 mM磷酸盐缓冲液（pH 6.6）或含有1.25 mM腐胺的100 mM Tricine缓冲液（pH 8.2），可获得携带杂合和复合型链的rt‐PA糖肽的高分辨率和可重现分离。根据唾液酸含量和结构，在20 min内分离胎球蛋白、t-PA和α1-酸性糖蛋白的N-寡聚体。使用寡糖指纹技术，如本发明的技术，可以在生物技术中具有许多应用，例如评估糖蛋白生产的一致性或用于分析糖蛋白化学。
Capillary zone electrophoresis (CZE) has been investigated as an alternative method to analyze the carbohydrate moieties of glycoproteins. Carbohydrate‐mediated microheterogeneity of the recombinant plasminogen activator (rt‐PA) was examined. The glycoprotein was resolved in multiple electrophoretic species using CZE but the separation was complicated by adsorption of the molecules to the wall of the capillary. The influence of several parameters, such as pH, molarity of the buffer and addition of a cationic additive, on the separation of glycopeptides was investigated. High resolution and reproducible separations of rt‐PA glycopeptides carrying hybrid and complex type chains were obtained using either a 100 mM phosphate buffer, pH 6.6, or a 100 mM Tricine buffer, pH 8.2, containing 1.25 mM of putrescine. N‐Oligosaccharides from fetuin, t‐PA and α1‐acid glycoprotein were separated within 20 min on the basis of both their sialic acid content and their structure. The use of an oligosaccharide fingerprinting technique, such as the present one, could have many applications in biotechnology to assess, for example, the consistency of production of a glycoprotein or for analytical glycoprotein chemistry.
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影响因子：
2.9
作者：
HARDY, MR;TOWNSEND, RR;LEE, YC
通讯作者：
LEE, YC
影响因子：
2.9
作者：
Raymond R. Townsend;M. Hardy;Dale A. Cumming;J. P. Carver;Brad Bendiak
通讯作者：
Raymond R. Townsend;M. Hardy;Dale A. Cumming;J. P. Carver;Brad Bendiak
影响因子：
2.9
作者：
Jentoft,N
通讯作者：
Jentoft,N
DOI：
10.1073/pnas.85.10.3289
发表时间：
1988-05-01
影响因子：
11.1
作者：
HARDY, MR;TOWNSEND, RR
通讯作者：
TOWNSEND, RR