Design of magnetic polyethyleneimine-fluorescein isothiocyanate composites toward efficient enrichment of phosphopeptides

IF 4 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A Pub Date : 2025-01-13 DOI:10.1016/j.chroma.2025.465679

Jinfeng Zhang, Yaming Zhao, Zirui Wang, Wanshu Wang, Jiutong Ma, Qiong Jia

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Abstract

Phosphoproteins maintain the normal metabolic activity of the organisms. Direct phosphopeptides detection is difficult to be realized by mass spectroscopy (MS) due to the low ionization efficiency, low abundance of phosphopeptides and interferences of complicated biological fluids. In the present work, a magnetic composite material was prepared by combining polyethyleneimine (PEI) and fluorescein isothiocyanate (FITC) focusing on phosphopeptides enrichment. Fourier-transform infrared spectroscopy, thermogravimetry analysis, X-ray photoemission spectroscopy, X-ray diffraction and UV–vis spectrometry measurement results confirmed the successful synthesis of Fe₃O₄@PEI@FITC material. FITC was facilely connected with PEI, and the two motifs endowed the efficient enrichment capacity of phosphopeptides by simple regulation of pH. The Fe₃O₄@PEI@FITC material was applied to the detection of phosphopeptides from real samples, implying that it can serve as an applicable platform for phosphoproteomics analysis.

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磁性聚乙烯亚胺-异硫氰酸荧光素复合材料高效富集磷酸肽的设计。

磷蛋白维持生物体正常的代谢活动。由于质谱法电离效率低、多肽丰度低以及复杂生物流体的干扰，难以实现直接的磷酸肽检测。本文以聚乙烯亚胺（PEI）和异硫氰酸荧光素（FITC）为主要富集材料，制备了一种磁性复合材料。傅里叶变换红外光谱、热重分析、x射线光发射光谱、x射线衍射和紫外可见光谱测量结果证实了Fe3O4@PEI@FITC材料的成功合成。FITC与PEI有很好的连接，这两个基序通过简单的ph调节就赋予了磷酸肽的高效富集能力。Fe3O4@PEI@FITC材料被应用于实际样品中磷酸肽的检测，这意味着它可以作为磷酸蛋白质组学分析的适用平台。

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来源期刊

Journal of Chromatography A 化学-分析化学

CiteScore

7.90

自引率

14.60%

发文量

742

审稿时长

45 days

期刊介绍： The Journal of Chromatography A provides a forum for the publication of original research and critical reviews on all aspects of fundamental and applied separation science. The scope of the journal includes chromatography and related techniques, electromigration techniques (e.g. electrophoresis, electrochromatography), hyphenated and other multi-dimensional techniques, sample preparation, and detection methods such as mass spectrometry. Contributions consist mainly of research papers dealing with the theory of separation methods, instrumental developments and analytical and preparative applications of general interest.