Semi-quantitative analysis of serum and cerebrospinal fluid transferrin glycoforms by top-down liquid chromatography mass spectrometry

IF 2.8 3区医学 Q2 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography B Pub Date : 2024-09-12 DOI:10.1016/j.jchromb.2024.124306

Richard K.T. Kam, Jenny Y.K. Cheng, Shreenidhi R. Subramaniam, Jeffrey S.S. Kwok

引用次数: 0

Abstract

Detection of β-2 transferrin in body fluid could help identify cerebrospinal fluid (CSF) leakage. The most common method, isoelectric focusing, was qualitative and could not provide detailed N-glycan structural information. We presented an alternative method using top-down liquid chromatography-time of flight mass spectrometry (LC-TOF MS). After immunoaffinity enrichment, fluid transferrin glycoforms were analyzed by a high-resolution LC-TOF MS, and the N-glycan structure predicted by accurate mass. The performance was validated with imprecision at 15%, with a cut-off of 0.04 for β-2 transferrin to tetrasialotransferrin ratio to confirm the presence of CSF in fluid samples.

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利用自上而下液相色谱质谱法对血清和脑脊液转铁蛋白糖形进行半定量分析

检测体液中的β-2转铁蛋白有助于鉴别脑脊液（CSF）渗漏。最常用的方法是等电聚焦法，这种方法只能定性，不能提供详细的 N-聚糖结构信息。我们提出了一种使用自上而下液相色谱-飞行时间质谱法（LC-TOF MS）的替代方法。免疫亲和富集后，用高分辨率液相色谱-飞行时间质谱分析流体转铁蛋白糖形，并通过精确质量预测 N-聚糖结构。该方法的性能得到了验证，不精确度为 15%，β-2 转铁蛋白与四硅转铁蛋白比值的临界值为 0.04，可用于确认液体样本中是否存在 CSF。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Chromatography B 医学-分析化学

CiteScore

5.60

自引率

3.30%

发文量

306

审稿时长

44 days

期刊介绍： The Journal of Chromatography B publishes papers on developments in separation science relevant to biology and biomedical research including both fundamental advances and applications. Analytical techniques which may be considered include the various facets of chromatography, electrophoresis and related methods, affinity and immunoaffinity-based methodologies, hyphenated and other multi-dimensional techniques, and microanalytical approaches. The journal also considers articles reporting developments in sample preparation, detection techniques including mass spectrometry, and data handling and analysis. Developments related to preparative separations for the isolation and purification of components of biological systems may be published, including chromatographic and electrophoretic methods, affinity separations, field flow fractionation and other preparative approaches. Applications to the analysis of biological systems and samples will be considered when the analytical science contains a significant element of novelty, e.g. a new approach to the separation of a compound, novel combination of analytical techniques, or significantly improved analytical performance.