Pradita Tiara, Chen Yi-Ju, Mernie Elias Gizaw, Bendulo Sharine Noelle, Chen Yu-Ju
Institute of Chemistry, Academia Sinica, Taipei 115, Taiwan.
Sustainable Chemical Science and Technology, Taiwan International Graduate Program, Academia Sinica, Taipei 115, Taiwan.
Nanomaterials (Basel). 2021 Aug 24;11(9):2159. doi: 10.3390/nano11092159.
Due to their unique glycan composition and linkage, protein glycosylation plays significant roles in cellular function and is associated with various diseases. For comprehensive characterization of their extreme structural complexity occurring in >50% of human proteins, time-consuming multi-step enrichment of glycopeptides is required. Here we report zwitterionic n-dodecylphosphocholine-functionalized magnetic nanoparticles (ZIC-cHILIC@MNPs) as a highly efficient affinity nanoprobe for large-scale enrichment of glycopeptides. We demonstrate that ZIC-cHILIC@MNPs possess excellent affinity, with 80-91% specificity for glycopeptide enrichment, especially for sialylated glycopeptide (90%) from biofluid specimens. This strategy provides rapidity (~10 min) and high sensitivity (<1 μL serum) for the whole enrichment process in patient serum, likely due to the rapid separation using magnetic nanoparticles, fast reaction, and high performance of the affinity nanoprobe at nanoscale. Using this strategy, we achieved personalized profiles of patients with hepatitis B virus (HBV, = 3) and hepatocellular carcinoma (HCC, = 3) at the depth of >3000 glycopeptides, especially for the large-scale identification of under-explored sialylated glycopeptides. The glycoproteomics atlas also revealed the differential pattern of sialylated glycopeptides between HBV and HCC groups. The ZIC-cHILIC@MNPs could be a generic tool for advancing the glycoproteome analysis, and contribute to the screening of glycoprotein biomarkers.
由于其独特的聚糖组成和连接方式，蛋白质糖基化在细胞功能中发挥着重要作用，并与多种疾病相关。为了全面表征超过50%的人类蛋白质中存在的极其复杂的结构，需要对糖肽进行耗时的多步富集。在此，我们报告了两性离子正十二烷基磷酸胆碱功能化的磁性纳米颗粒（ZIC-cHILIC@MNPs），作为一种用于大规模富集糖肽的高效亲和纳米探针。我们证明ZIC-cHILIC@MNPs具有出色的亲和力，对糖肽富集的特异性为80 - 91%，特别是对生物流体样本中的唾液酸化糖肽（90%）。该策略为患者血清中的整个富集过程提供了快速性（约10分钟）和高灵敏度（<1 μL血清），这可能是由于使用磁性纳米颗粒进行快速分离、快速反应以及亲和纳米探针在纳米尺度上的高性能。使用该策略，我们在超过3000个糖肽的深度上实现了乙型肝炎病毒（HBV，n = 3）和肝细胞癌（HCC，n = 3）患者的个性化图谱，特别是用于大规模鉴定未充分探索的唾液酸化糖肽。糖蛋白质组图谱还揭示了HBV和HCC组之间唾液酸化糖肽的差异模式。ZIC-cHILIC@MNPs可能是推进糖蛋白质组分析的通用工具，并有助于糖蛋白生物标志物的筛选。
