Upconversion Nanoparticles-Encoded Hydrogel Microbeads-Based Multiplexed Protein Detection.
Upconversion Nanoparticles-Encoded Hydrogel Microbeads-Based Multiplexed Protein Detection.
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DOI：
10.1007/s40820-017-0184-y
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发表时间：
2018
影响因子：
26.6
通讯作者：
Zhang Y
中科院分区：
文献类型：
--
作者：
Shikha S;Zheng X;Zhang Y
重复编码的微珠在不同领域的多重应用中是有需求的。与基于有机染料的市售Luminex的xMAP技术相比，上转换纳米颗粒（UCNP）是更好的替代品，因为它们具有大的反斯托克斯位移，光稳定性，零背景和单波长激发。在这里，我们开发了一种新的多重检测系统，使用UCNPs编码聚（乙二醇）二丙烯酸酯（PEGDA）微珠以及标记报告抗体。然而，为了制备UCNP编码的微珠，目前使用的基于溶胀的包封导致不均匀性，这对于基于荧光的多路复用是不期望的。因此，我们利用液滴微流控技术来获得均匀大小、形状和内部UCNP分布的编码微珠。此外，PEGDA微珠缺乏在其表面上用于探针抗体缀合的功能。迄今报道的PEGDA微珠表面官能化的方法（丙烯酸掺入，聚多巴胺涂层）淬灭UCNP的荧光。在这里，PEGDA微珠表面涂覆有二氧化硅，然后进行羧基修饰，而不损害UCNP的荧光强度。在这项研究中，液滴微流体辅助UCNPs编码的微珠的形状，大小和荧光均匀的制备。通过在包封之前以不同比例混合发射红色和绿色的UCNP来产生多种颜色代码。发射蓝色的UCNP用于标记报告抗体。探针抗体被共价固定在红色UCNPs编码的微珠上，用于特异性捕获作为模型蛋白的人血清白蛋白（HSA）。通过将抗hCRP抗体固定在绿色UCNP上，该系统还被证明用于人C-反应蛋白（hCRP）和HSA蛋白的多重检测。本文的在线版本（10.1007/s40820-017-0184-y）包含补充材料，可供授权用户使用。
Fluorescently encoded microbeads are in demand for multiplexed applications in different fields. Compared to organic dye-based commercially available Luminex’s xMAP technology, upconversion nanoparticles (UCNPs) are better alternatives due to their large anti-Stokes shift, photostability, nil background, and single wavelength excitation. Here, we developed a new multiplexed detection system using UCNPs for encoding poly(ethylene glycol) diacrylate (PEGDA) microbeads as well as for labeling reporter antibody. However, to prepare UCNPs-encoded microbeads, currently used swelling-based encapsulation leads to non-uniformity, which is undesirable for fluorescence-based multiplexing. Hence, we utilized droplet microfluidics to obtain encoded microbeads of uniform size, shape, and UCNPs distribution inside. Additionally, PEGDA microbeads lack functionality for probe antibodies conjugation on their surface. Methods to functionalize the surface of PEGDA microbeads (acrylic acid incorporation, polydopamine coating) reported thus far quench the fluorescence of UCNPs. Here, PEGDA microbeads surface was coated with silica followed by carboxyl modification without compromising the fluorescence intensity of UCNPs. In this study, droplet microfluidics-assisted UCNPs-encoded microbeads of uniform shape, size, and fluorescence were prepared. Multiple color codes were generated by mixing UCNPs emitting red and green colors at different ratios prior to encapsulation. UCNPs emitting blue color were used to label the reporter antibody. Probe antibodies were covalently immobilized on red UCNPs-encoded microbeads for specific capture of human serum albumin (HSA) as a model protein. The system was also demonstrated for multiplexed detection of both human C-reactive protein (hCRP) and HSA protein by immobilizing anti-hCRP antibodies on green UCNPs. The online version of this article (10.1007/s40820-017-0184-y) contains supplementary material, which is available to authorized users.
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