Separation of oligo/polymers of 5-N-acetylneuraminic acid, 5-N-glycolylneuraminic acid, and 2-keto-3-deoxy-D-glycero-D-galacto-nononic acid by high-performance anion-exchange chromatography with pulsed amperometric detector.
Separation of oligo/polymers of 5-N-acetylneuraminic acid, 5-N-glycolylneuraminic acid, and 2-keto-3-deoxy-D-glycero-D-galacto-nononic acid by high-performance anion-exchange chromatography with pulsed amperometric detector.
复制标题
采用脉冲电流分析法的高性能阴离子交换色谱法分离 5-N-乙酰神经氨酸、5-N-羟乙酰神经氨酸和 2-酮-3-脱氧-D-甘油-D-半乳糖壬酸的低聚物/聚合物
DOI：
10.1006/abio.1997.2225
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发表时间：
1997
影响因子：
2.9
通讯作者：
Lee,YC
中科院分区：
文献类型：
--
作者：
Zhang,Y;Inoue,Y;Inoue,S;Lee,YC
开发了一种灵敏、高效的方法，使用高效阴离子交换色谱法（HPAEC）和脉冲电流检测器（PAD-2）分析含有α2-8连接的5-N-乙酰神经氨酸（Neu 5Ac）、5-N-羟乙酰神经氨酸（Neu 5Gc）和脱氨神经氨酸（KDN）的低聚/多聚唾液酸。使用CarboPac PA-100色谱柱和硝酸钠作为推进剂，在68 min内分离了聚合度（DP）高达80的多聚体。在CarboPac PA-1色谱柱上也获得了类似的DP为基础的分辨率。Neu 5Ac、Neu 5Gc和KDN系列的洗脱阶梯差异很大，足以用作诊断指标。该技术被应用于鉴定的唾液酸组分中的聚唾液酸糖蛋白（PSGP）样品，以及监测的寡/多聚-KDN-含馏分在纯化过程中的含KDN-gp（KDN-gp）。在PSGP和KDN-gp水解产物中可检测到的oligo-Neu 5Gc和oligo-KDN的最大DP分别为11和8。通过定量Neu 5Ac寡聚体证明了该方法的高灵敏度。研究了在不同条件下，多聚体和多聚体在多聚酸和多聚PSGP的酸解产物和酶解产物中的分布。
A sensitive and efficient method to analyze oligo/poly-sialic acids containing α2–8-linked 5-N-acetylneuraminic acid (Neu5Ac), 5-N-glycolylneuraminic acid (Neu5Gc), and deaminated neuraminic acid (KDN) using high-performance anion-exchange chromatography (HPAEC) with a pulsed amperometric detector (PAD-2) has been developed. Using a CarboPac PA-100 column and sodium nitrate as the pushing agent, polymers in colominic acid with degree of polymerization (DP) up to 80 were separated in 68 min. A similar DP-based resolution was also obtained on a CarboPac PA-1 column. The elution ladders of the Neu5Ac, Neu5Gc, and KDN series were sufficiently different to be used as diagnostic indices. This technique was applied to identification of the sialic acid components in a polysialoglycoprotein (PSGP) sample as well as monitoring the oligo/poly-KDN-containing fractions during the purification of KDN-containing glycoprotein (KDN-gp). The maximum DPs of oligo-Neu5Gc and oligo-KDN that can be detected in PSGP and KDN-gp hydrolysates were 11 and 8, respectively. The high sensitivity of this method was demonstrated by the quantification of Neu5Ac oligomers. Distributions of the monomer and oligo/polymers in the acid and enzymatic hydrolysates of colominic acid and PSGP under different conditions were also studied.