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Analysis method and analysis system for N-linked sugar chain of glycoprotein

机译:糖蛋白N联糖链的分析方法和分析系统

摘要

Disclosed is a novel means for accurate qualitative and quantitative analyses for each N-glycosylation site. The method of analyzing N-linked sugar chain(s) of glycoprotein according to the present invention comprises: treating a part of a glycopeptide-containing sample to be analyzed with endo-β-N-acetylglucosaminidases to cleave off sugar chains while leaving one GlcNAc of the chitobiose core on the Asn at the N-glycosylation site; subjecting the obtained sugar chain-cleaved sample to preliminary liquid chromatography/mass spectrometry; predicting the retention time of the glycopeptide of interest and the mass-to-charge ratio (m/z) of the precursor ion in main analysis based on the results of the preliminary liquid chromatography/mass spectrometry; and carrying out the main analysis. By this method, the binding sites and structures of N-linked sugar chains in a glycoprotein can be analyzed. By using the sugar chain-cleaved sample as an internal standard in the main analysis, quantitative analysis of sugar chains at each glycosylation site also becomes possible.
机译:公开了一种用于对每个N-糖基化位点进行准确的定性和定量分析的新颖方法。根据本发明的分析糖蛋白的N-连接糖链的方法包括:用β-N-乙酰基氨基葡萄糖内切酶处理待分析的含糖肽样品的一部分,以切割掉糖链,同时留下一个GlcNAc。在N-糖基化位点的Asn上的壳二糖核心;将获得的糖链切割的样品进行初步液相色谱/质谱分析;根据初步液相色谱/质谱法的结果,预测主要分析中目标糖肽的保留时间和前体离子的质荷比(m / z);并进行主要分析。通过这种方法,可以分析糖蛋白中N-连接糖链的结合位点和结构。通过在主分析中使用糖链裂解的样品作为内标,还可以对每个糖基化位点的糖链进行定量分析。

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