Extraction, Detection, and Profiling of Serum Biomarkers Using Designed Fe3O4@SiO2@HA Core-Shell Particles
Extraction, Detection, and Profiling of Serum Biomarkers Using Designed Fe3O4@SiO2@HA Core-Shell Particles
复制标题
使用设计的 Fe3O4@SiO2@HA 核壳颗粒提取、检测和分析血清生物标志物
DOI：
10.1007/s12274-017-1591-6
复制
发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Qian Kun
中科院分区：
文献类型：
--
作者：
Rejeeth C;Pang X;Zhang R;Xu W;Sun X;Liu B;Lou J;Wan J;Gu H;Yan W;Qian Kun
蛋白质形式的血清生物标志物（如CD44）已被证明对疾病诊断和预后具有很高的临床敏感性和特异性。由于血清中样品的高复杂性和低分子丰度，生物标志物的检测和分析依赖于材料和设备的高效提取，主要使用抗体-抗原识别的免疫测定。无抗体的方法很有前途，需要开发用于血清的实际应用，以解决基于抗体的技术在稳健性、费用和吞吐量方面的局限性。在这项工作中，我们展示了一种利用透明质酸（HA）修饰的材料/设备通过配体-蛋白相互作用提取、检测和分析血清生物标志物的新方法。我们通过逐层组装构建了不同大小的Fe3O4@SiO2@HA颗粒，并首次将ha功能化颗粒应用于血清CD44的质谱快速提取和序列鉴定。我们还首次通过电化学传感验证了HA-CD44结合，使用ha修饰电极在标准溶液和稀释的血清样品中进行检测，检测限为~ 0.6 ng/mL，线性响应范围为1 ng/mL至10 μg/mL。此外，我们进行了ha结合血清蛋白质组的分析，为未来数据库的构建提供了新的初步基准，我们研究了用于捕获血清蛋白质的颗粒的表面化学性质。我们的工作不仅开发了CD44提取/检测和ha结合蛋白质组鉴定的平台技术，而且还指导了基于配体亲和的方法的设计，用于血清生物标志物的无抗体分析，以用于诊断应用。
Serum biomarkers in the form of proteins (e.g. cluster of differentiation-44 (CD44)) have been demonstrated to have high clinical sensitivity and specificity for disease diagnosis and prognosis. Owing to the high sample complexity and low molecular abundance in serum, the detection and profiling of biomarkers rely on efficient extraction by materials and devices, mostly using immunoassays via antibody-antigen recognition. Antibody-free approaches are promising and need to be developed for real-case applications in serum to address the limitations of antibody-based techniques in terms of robustness, expense, and throughput. In this work, we demonstrated a novel approach using hyaluronic acid (HA)-modified materials/devices for the extraction, detection, and profiling of serum biomarkers via ligand-protein interactions. We constructed Fe3O4@SiO2@HA particles with different sizes through layer-by-layer assembly and for the first time applied HA-functionalized particles in the facile extraction and sequence identification of CD44 in serum by mass spectrometry. We also first validated HA-CD44 binding through electrochemical sensing using HA-modified electrodes in both standard solutions and diluted serum samples, achieving a detection limit of ∼0.6 ng/mL and a linear response range from 1 ng/mL to 10 μg/mL. Furthermore, we performed profiling of HA-binding serum proteome, providing a new preliminary benchmark for the construction of future databases, and we investigated selected surface chemistries of particles for the capture of proteins in serum. Our work not only resulted in the development of a platform technology for CD44 extraction/detection and HA-binding proteome identification, but also guided the design of ligand affinity-based approaches for antibody-free analysis of serum biomarkers towards diagnostic applications.