Changes in the levels and compositions of N-glycans released from serum and plasma glycoproteins have been assessed in many diseases across many large clinical sample cohorts. Assays used for N-glycan profiling in these fluids currently require multiple processing steps and have limited throughput, thus diminishing their potential for use as standard clinical diagnostic assays. A novel slide-based N-glycan profiling method was evaluated for sensitivity and reproducibility using a pooled serum standard. Serum was spotted on to an amine-reactive slide, delipidated and desalted with a series of washes, sprayed with peptide N-glycosidase F and matrix, and analyzed by MALDI-FTICR or MALDI-Q-TOF mass spectrometry. Routinely, over 75 N-glycan species can be detected from one microliter of serum in less than 6.5 h. Additionally, endoglycosidase F3 was applied to this workflow to identify core-fucosylated N-glycans and displayed the adaptability of this method for the determination of structural information. This method was applied to a small pooled serum set from either obese or nonobese patients that had breast cancer or a benign lesion. This study confirms the reproducibility, sensitivity, and adaptability of a novel method for N-glycan profiling of serum and plasma for potential application to clinical diagnostics.

中文翻译：

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通过新颖的基于载玻片的成像质谱工作流程对血清和血浆进行快速 N-聚糖分析。

血清和血浆糖蛋白释放的 N-聚糖水平和成分的变化已在许多大型临床样本队列中的许多疾病中进行了评估。目前，用于这些液体中 N-聚糖分析的测定需要多个处理步骤，并且通量有限，因此削弱了它们作为标准临床诊断测定的潜力。使用混合血清标准品评估了一种基于载玻片的新型 N-聚糖分析方法的灵敏度和重现性。将血清点样到胺反应性载玻片上，通过一系列洗涤进行脱脂和脱盐，喷洒肽 N-糖苷酶 F 和基质，并通过 MALDI-FTICR 或 MALDI-Q-TOF 质谱进行分析。通常，在 6.5 小时内可以从 1 微升血清中检测到超过 75 种 N-聚糖。此外，糖苷内切酶 F3 应用于该工作流程来识别核心岩藻糖基化 N-聚糖，并展示了该方法对于确定结构信息的适应性。该方法适用于来自患有乳腺癌或良性病变的肥胖或非肥胖患者的少量混合血清组。这项研究证实了一种用于血清和血浆 N-聚糖分析的新方法的重现性、灵敏度和适应性，有可能应用于临床诊断。