Microfluidic-based multiplex immunoassay system integrated with an array of QD-encoded microbeads
Microfluidic-based multiplex immunoassay system integrated with an array of QD-encoded microbeads
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DOI：
10.1016/j.snb.2014.11.115
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发表时间：
2015-03-31
影响因子：
8.4
通讯作者：
Koh, Won-Gun
中科院分区：
文献类型：
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作者：
Han, Sang Won;Jang, Eunji;Koh, Won-Gun
在这里，我们开发了一个多重免疫分析平台内的微流控装置，结合悬浮液和平面微阵列格式。对于悬浮液微阵列格式，QD嵌入的聚合物微珠的平均直径为24 μ m，使用Shirasu多孔玻璃（SPG）膜乳化技术制备。为了给微珠提供可分辨的光谱代码，使用具有两种不同颜色（450 nm用于蓝色和520 nm用于绿色）的QD，并且通过改变微珠内两种不同QD的发射强度的比率来获得不同的光谱代码。然后用探针抗体将QD编码的微珠的表面功能化用于免疫测定。通过在PDMS中制造的微孔阵列实现平面微阵列格式。每个微孔被设计成捕获单个微珠，并最终在微流体系统内产生微珠阵列。通过IgG和抗IgG之间和/或IgM和抗IgM之间的免疫结合测定证明了微流体装置内的微珠阵列用于多重免疫测定的可行性。此外，所得到的基于微珠的芯片上测定可用于夹心测定以检测前列腺特异性抗原（PSA）（一种模型癌症标志物），检测限为1 ng/mL。悬浮液和平面微阵列格式的组合使得单个微珠在物理上分离的区域内的空间定位成为可能，从而促进了与相应微珠相互作用的不同靶标的同时测定。(C)2014年爱思唯尔B。V.保留所有权利。
Here, we developed a multiplex immunoassay platform within microfluidic devices that combines suspension and the planar microarray format. For the suspension microarray format, QD-embedded polymeric microbeads with an average diameter of 24 mu m were prepared using the Shirasu Porous Glass (SPG) membrane emulsification technique. To furnish the microbeads with resolvable spectral codes, QDs with two different colors (450 nm for blue and 520 nm for green) were used and different spectral codes were obtained by changing the ratio of emission intensity of the two different QDs within the microbeads. The surfaces of the QD-encoded microbeads were then functionalized with probe antibodies for immunoassays. The planar microarray format was achieved by an array of microholes fabricated in PDMS. Each microhole was designed to trap a single microbead and eventually generated a microbeads array within the microfluidic system. The feasibility of the microbead array within microfluidic devices for use in a multiplex immunoassay was demonstrated by immunobinding assays between IgG and anti IgG and/or between IgM and anti-IgM. Furthermore, the resultant microbead-based on-chip assay could be used for a sandwich assay to detect prostate-specific antigen (PSA), a model cancer marker, with a detection limit of 1 ng/mL. The combination of suspension and the planar microarray format enabled the spatial location of individual microbeads within physically separated regions and thus facilitated the simultaneous determination of different targets that interacted with the corresponding microbeads. (C) 2014 Elsevier B. V. All rights reserved.