色谱

色谱 ›› 2021, Vol. 39 ›› Issue (3): 205-210.DOI: 10.3724/SP.J.1123.2020.11006

• 研究快报 • 上一篇    下一篇

两性离子双功能化超亲水金属有机框架纳米复合材料的制备及其用于糖肽的选择性富集

李大鹏, 谢光珊, 谢佩斯, 朱林, 蔡宗苇*()   

  1. 香港浸会大学, 环境与生物分析国家重点实验室, 香港 999077
  • 收稿日期:2020-11-06 出版日期:2021-03-08 发布日期:2021-02-03
  • 通讯作者: 蔡宗苇
  • 作者简介:*Tel:(852)34117070,E-mail: zwcai@hkbu.edu.hk.
  • 基金资助:
    国家自然科学基金重点计划(2017YFC1600505);香港、澳门和台湾地区科技创新合作国家重点研究开发计划(2017YFE0191000)

Synthesis of zwitterionic dual-functional metal-organic framework nanocomposite with ultra-hydrophilicity for selective enrichment of glycopeptides

LI Dapeng, XIE Guangshan, XIE Peisi, ZHU Lin, CAI Zongwei*()   

  1. State Key Laboratory of Environmental and Biological Analysis, Hong Kong Baptist University, Hong Kong 999077, China
  • Received:2020-11-06 Online:2021-03-08 Published:2021-02-03
  • Contact: CAI Zongwei
  • Supported by:
    Key Program of the National Natural Science Foundation of China(2017YFC1600505);National Key Research and Development Program-Cooperation on Scientific and Technological Innovation in Hong Kong, Macao and Taiwan(2017YFE0191000)

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摘要：

蛋白糖基化是生物体中普遍发生且重要的生物学过程,其参与多种分子生物学的功能和途径,是临床诊断重要的生物标志物。但是,糖肽因其丰度低、离子化效率低、糖链异质性等难点,使糖蛋白分析一直面临巨大的挑战。因此,研究合成了一种新型的两性离子双功能化纳米金(AuGC)修饰的超亲水性沸石咪唑骨架(ZIF-8)纳米复合材料(AuGC/ZIF-8),并建立了亲水相互作用色谱(HILIC)和基质辅助激光解吸电离-飞行时间质谱(MALDI-TOF MS)联用选择性富集糖肽的分析方法。谷胱甘肽和半胱氨酸双功能化的协同作用,使MOF具有超亲水性和低空间位阻,为糖肽选择性富集提供了更多的亲和位点。研究以辣根过氧化物酶(HRP)为模式糖蛋白,通过AuGC/ZIF-8富集后,MALDI-TOF MS分析。结果表明,AuGC/ZIF-8对HRP糖肽的富集能力高达250 μg/mg,且在与牛血清白蛋白(BSA)混合溶液中(HRP-BSA (1∶200,质量比))显示出对HRP糖肽的高选择性,以及极低含量下(0.3 ng/μL)的高灵敏度。因此,在复杂生物样品糖蛋白的富集分离中具有很大的应用潜力。

关键词: 金属有机框架, 亲水相互作用色谱, 基质辅助激光解吸电离-飞行时间质谱, 选择性富集, 翻译后修饰, 糖肽

Abstract:

Protein glycosylation is a ubiquitous and important biological process involved in various molecular functions and biological pathways. It also yields important biomarkers for clinical diagnoses. However, glycopeptide analysis is challenging due to low abundance, low ionization efficiency, and glycan heterogeneity. In the present study, a method based on hydrophilic interaction liquid chromatography (HILIC) was developed for the selective enrichment of glycopeptides using a novel metal-organic framework (MOF) nanocomposite (AuGC/ZIF-8). Dual functionalization with glutathione and cysteine has resulted in an ultra-hydrophilic MOF, with synergistic effects and lower steric hindrance, providing more affinity sites for the glycopeptide enrichment. Horseradish peroxidase (HRP) was used as a model glycoprotein, and AuGC/ZIF-8 was used to enrich glycopeptides prior to analysis by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). AuGC/ZIF-8 displayed outstanding performance at enriching HRP glycopeptides, with high enrichment capacity (250 μg/mg), high selectivity in mixtures containing bovine serum albumin (BSA) (HRP-BSA (1∶200, mass ratio)), and high sensitivity at very low content (0.3 ng/μL). Thus this MOF holds promise for in-depth, comprehensive glycoproteomic and related analysis.

Key words: metal-organic framework (MOF), hydrophilic interaction liquid chromatography (HILIC), matrix-assisted laser desorption ionization time-of-flight-mass spectrometry (MALDI-TOF MS), selective enrichment, post-translational modification (PTM), glycopeptide

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