Dua V K, Bush C A
Anal Biochem. 1984 Feb;137(1):33-40. doi: 10.1016/0003-2697(84)90342-7.
Reverse-phase high-pressure liquid chromatography has been shown to be useful in fractionation of oligosaccharides containing N-acetyl amino sugars on the basis of stereochemical differences. Data in this paper show this same stereochemical selectivity can be obtained for glycopeptides. Using dilute aqueous phosphate for elution, three major high-mannose glycopeptides from ovalbumin can be separated on the analytical scale in a few minutes and the same method can also be used for preparation of the glycopeptides on the milligram scale. High-field proton NMR spectroscopy is used to identify two of the compounds as the mannose-5 and -6 glycopeptides (E3 and D3) and the third as the mannose-6 glycopeptide substituted by two N-acetyl glucosamine (GlcNAc) residues (C3B). The NMR spectra and analytical chromatograms of the glycopeptides show high purity and freedom from contaminants previously identified in ovalbumin glycopeptides.
反相高效液相色谱已被证明可用于基于立体化学差异对含N-乙酰氨基糖的寡糖进行分级分离。本文中的数据表明，对于糖肽也可获得相同的立体化学选择性。使用稀磷酸盐水溶液进行洗脱，在几分钟内即可在分析规模上分离出卵清蛋白中的三种主要高甘露糖糖肽，并且相同的方法也可用于毫克级糖肽的制备。高场质子核磁共振光谱用于鉴定其中两种化合物为甘露糖-5和-6糖肽（E3和D3），第三种为被两个N-乙酰葡糖胺（GlcNAc）残基取代的甘露糖-6糖肽（C3B）。糖肽的核磁共振光谱和分析色谱图显示出高纯度且不含先前在卵清蛋白糖肽中鉴定出的污染物。
