Analysis of the N-acetylneuraminic acid and N-glycolylneuraminic acid contents of glycoproteins by high-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC/PAD)
Analysis of the N-acetylneuraminic acid and N-glycolylneuraminic acid contents of glycoproteins by high-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC/PAD)
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DOI：
10.1093/glycob/8.1.35
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发表时间：
1998-01-01
期刊：
影响因子：
4.3
通讯作者：
Avdalovic, N
中科院分区：
文献类型：
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作者：
Rohrer, JS;Thayer, J;Avdalovic, N
N-乙酰神经氨酸 (Neu5Ac) 的存在或不存在会改变唾液酸化糖蛋白的血清半衰期及其功能，我们评估了 HPAEC/PAD 方法的线性、灵敏度、重现性和准确性，以确定其是否适合对 Neu5Ac 和 N-羟乙酰神经氨酸 (Neu5Gc) 进行常规同步分析。此分析的有效内标是3-脱氧-d-甘油-d-半乳糖-2-壬糖酸（KDN）。我们研究了 Au 工作电极衰退的影响，并确定线性范围和灵敏度取决于电极衰退，使用距电极块凹进 350 μm 的电极，Neu5Ac、KDN 和 Neu5Gc 的最小检测限分别为 2、5 和 2 pmol，使用新电极降低至 1、2 和 0.5 pmol，标准品的响应从 10 到 500 pmol 呈线性(r(2) > 0.99) 无论电极衰退如何，当重复分析 Neu5Ac、KDN 和 Neu5Gc（各 200 pmol）48 小时时，面积 RSD 为
Presence or absence of N-acetylneuraminic acid (Neu5Ac) can change a sialylated glycoprotein's serum half-life and possibly its function, We evaluated the linearity, sensitivity, reproducibility, and accuracy of a HPAEC/PAD method to determine its suitability for routine simultaneous analysis of Neu5Ac and N-glycolylneuraminic acid (Neu5Gc), An effective internal standard for this analysis is 3-deoxy-d-glycero-d-galacto-2-nonulosonic acid (KDN). We investigated the effect of the Au working electrode recession and determined that linear range and sensitivity were dependent on electrode recession, Using an electrode that was 350 mu m recessed from the electrode block, the minimum detection limits of Neu5Ac, KDN, and Neu5Gc were 2, 5, and 2 pmol, respectively, and were reduced to 1, 2, and 0.5 pmol using a new electrode, The response of standards was linear from 10 to 500 pmol (r(2) > 0.99) regardless of electrode recession, When Neu5Ac, KDN, and Neu5Gc (200 pmol each) were analyzed repetitively for 48 h, area RSDs were