Beijing Key Laboratory of Environmentally Harmful Chemical Analysis, College of Chemistry, Beijing University of Chemical Technology, Beijing 100029, China.
Beijing National Laboratory for Molecular Sciences (BNLMS), Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Analytical Instrumentation Center, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Jan 1;1232:123920. doi: 10.1016/j.jchromb.2023.123920. Epub 2023 Nov 15.
Protein glycosylation research is currently focused on the development of various functionalized materials that can effectively enrich the levels of glycopeptides in samples. However, most of these materials possess limited glycopeptide-specific recognition sites because of large steric hindrance, unsuitable mass transfer kinetics, and relatively low surface areas. Herein, a highly hydrophilic two-dimensional (2-D) metal-organic framework (MOF) nanosheet modified with glutathione (GSH) and l-cysteine (l-Cys) (denoted as Zr-Fc MOF@Au@GC) has been synthesized for efficient glycopeptide enrichment. Using this composite material, 39 and 44 glycopeptides from horseradish peroxidase (HRP) and human serum immunoglobulin G (IgG) digests were detected, respectively, which represents a higher efficiency for glycopeptide enrichment from model glycoprotein digests than has been previously reported. The material Zr-Fc MOF@Au@GC exhibited ultra-high sensitivity (0.1 fmol/µL), excellent selectivity (weight ratio of HRP tryptic digest to bovine serum albumin (BSA) tryptic digest = 1:2000), good binding capacity (200 mg/g), satisfactory reusability, and long-term storage capacity. In addition, 655 glycopeptides corresponding to 366 glycoproteins were identified from human serum samples. To the best of our knowledge, this is the largest number of glycoproteins detected in human serum samples to date. These results indicated that Zr-Fc MOF@Au@GC has the potential to be used for the enrichment of glycopeptides in biological samples and the analysis of protein glycosylation.
蛋白质糖基化研究目前集中于开发各种功能化材料，这些材料可以有效地提高样品中糖肽的水平。然而，由于较大的空间位阻、不合适的传质动力学和相对较低的表面积，这些材料大多具有有限的糖肽特异性识别位点。在此，合成了一种高度亲水性的二维（2-D）金属有机骨架（MOF）纳米片，该纳米片经谷胱甘肽（GSH）和半胱氨酸（l-Cys）修饰（记为 Zr-Fc MOF@Au@GC），用于高效糖肽富集。使用这种复合材料，分别从辣根过氧化物酶（HRP）和人血清免疫球蛋白 G（IgG）消化物中检测到 39 个和 44 个糖肽，这代表了从模型糖蛋白消化物中进行糖肽富集的效率高于之前的报道。Zr-Fc MOF@Au@GC 材料表现出超高的灵敏度（0.1 fmol/µL）、优异的选择性（HRP 酶解物与牛血清白蛋白（BSA）酶解物的重量比 = 1:2000）、良好的结合能力（200 mg/g）、令人满意的可重复使用性和长期储存能力。此外，从人血清样品中鉴定出 655 个糖肽对应 366 种糖蛋白。据我们所知，这是迄今为止在人血清样品中检测到的糖蛋白数量最多的一次。这些结果表明，Zr-Fc MOF@Au@GC 有可能用于生物样品中糖肽的富集和蛋白质糖基化的分析。
