State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, College of Chemistry, Nankai University, Tianjin, 300071, China.
State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, College of Chemistry, Nankai University, Tianjin, 300071, China.
Anal Chim Acta. 2020 May 1;1109:53-60. doi: 10.1016/j.aca.2020.02.066. Epub 2020 Mar 3.
Protein glycosylation is an important post-translational modification and glycoproteins are associated with many crucial metabolic progresses of life. In order to detect glycoproteins sensitively, we propose a gold nanoparticles (GNPs) enumeration method based on boronate affinity sandwich system, which is constructed between the boronic acid polymer functionalized magnetic nanoparticles (FeO@MPS@VPBA NPs) and 4-mercaptophenylboronic acid modified GNPs (GNPs-MPBA) by the targeted glycoproteins as the linker. Therefore, the sandwich complex is formed, resulting in the decrease of GNPs-MPBA counts in the solution. Based on the dark-field microscope (DFM) imaging technique, the sensitive GNPs enumeration assay is developed for glycoproteins quantitation. Immunoglobulin (IgG), as one of the important glycoproteins, is introduced to evaluate the proposed method. A low detection limit of 1.22 ng mL for IgG analysis is obtained. The result indicates that the proposed GNPs enumeration method offers a simple, effective, label-free and highly sensitive strategy without signal amplification. It also possesses great potential for various target molecules determination at the single-particle level in the future.
蛋白质糖基化是一种重要的翻译后修饰，糖蛋白与生命的许多关键代谢过程有关。为了灵敏地检测糖蛋白，我们提出了一种基于硼酸亲和三明治体系的金纳米粒子（GNPs）计数方法，该体系由硼酸盐聚合物功能化磁性纳米粒子（FeO@MPS@VPBA NPs）和 4-巯基苯硼酸修饰的 GNPs（GNPs-MPBA）通过作为连接物的靶向糖蛋白构建而成。因此，形成了三明治复合物，导致溶液中 GNPs-MPBA 的数量减少。基于暗场显微镜（DFM）成像技术，开发了用于糖蛋白定量的灵敏 GNPs 计数测定法。免疫球蛋白（IgG）作为一种重要的糖蛋白，被引入来评估所提出的方法。对于 IgG 分析，检测限低至 1.22ng/mL。结果表明，所提出的 GNPs 计数方法提供了一种简单、有效、无标记且高度灵敏的策略，无需信号放大。它还有望在未来用于在单颗粒水平上测定各种靶分子。
