Glucose unit index (GUI) of permethylated glycans for effective identification of glycans and glycan isomers.
Glucose unit index (GUI) of permethylated glycans for effective identification of glycans and glycan isomers.
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DOI：
10.1039/d0an00314j
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发表时间：
2020-10-21
期刊：
影响因子：
--
通讯作者：
Mechref Y
中科院分区：
文献类型：
--
作者：
Gautam S ;Peng W ;Cho BG ;Huang Y ;Banazadeh A ;Yu A ;Dong X ;Mechref Y
保留时间是使用液相色谱（LC）报告聚糖分离最常见和广泛使用的标准，但它在不同的柱，仪器和实验室中差异很大。当比较实验室间的数据时，这种差异是有问题的。此外，它影响再现性并妨碍有效的数据解释。为了克服这种差异，我们建议使用葡萄糖单位指数（GUI）对C18和PGC柱为基础的还原和过甲基化聚糖的分离。GUI以前被用于原生和标记聚糖的保留时间标准化。我们用从模型糖蛋白fetuin和核糖核酸酶B （RNase B）中提取的还原和过甲基化聚糖对该方法进行了评估，然后将其应用于人血清，生成C18和PGC柱基异构聚糖文库。根据糊精衍生的葡萄糖单位计算聚糖组成的GUI值，糊精被用作洗脱标准。在两个实验室的三种不同的LC系统（UltiMate 3000 Nano UHPLC系统）上验证GUI值，以确保方法的可靠性和可重复性。使用人类乳腺癌细胞系证明了对真实样品的适用性。通过C18柱分离得到116个过甲基化n -聚糖，通过PGC柱分离得到134个过甲基化n -聚糖。总的来说，建立的GUI方法和可重复的实验室间和实验室内GUI值的演示将有助于未来自动聚糖和异构聚糖鉴定方法的发展。
Retention time is the most common and widely used criterion to report the separation of glycans using Liquid Chromatography (LC), but it varies widely across different columns, instruments and laboratories. This variation is problematic when inter-laboratory data is compared. Furthermore, it influences reproducibility and hampers efficient data interpretation. In our endeavor to overcome this variance, we propose the use of the Glucose Unit Index (GUI) on C18 and PGC column-based separation of reduced and permethylated glycans. GUI has previously been utilized for retention time normalization of native and labeled glycans. We evaluated this method with reduced and permethylated glycans derived from model glycoproteins fetuin and ribonuclease B (RNase B), and then implemented it to human blood serum to generate C18 and PGC column-based isomeric glycan libraries. GUI values for glycan compositions were calculated with respect to the glucose units derived from dextrin, which was employed as an elution standard. The GUI values were validated on three different LC systems (UltiMate 3000 Nano UHPLC systems) in two laboratories to ensure the reliability and reproducibility of the method. Applicability on real samples was demonstrated using human breast cancer cell lines. A total of 116 permethylated N-glycans separated on a C18 column and 134 glycans separated on a PGC column were compiled in a library. Overall, the established GUI method and the demonstration of reproducible inter- and intra-laboratory GUI values would aid the future development of automated glycan and isomeric glycan identification methods.
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DOI：
10.1007/s13361-018-1933-y
发表时间：
2018-06
影响因子：
3.2
作者：
Benktander JD;Gizaw ST;Gaunitz S;Novotny MV
通讯作者：
Novotny MV
影响因子：
2.9
作者：
Guttman, A
通讯作者：
Guttman, A
影响因子：
7.4
作者：
Dong, Xue;Peng, Wenjing;Mechref, Yehia
通讯作者：
Mechref, Yehia
影响因子：
1.9
作者：
Gudelj, Ivan;Lauc, Gordan
通讯作者：
Lauc, Gordan
影响因子：
4.8
作者：
Lloyd, KO;Burchell, J;TaylorPapadimitriou, J
通讯作者：
TaylorPapadimitriou, J