Functionalized Polymer Microgel Particles Enable Customizable Production of Label-Free Sensor Arrays.
Functionalized Polymer Microgel Particles Enable Customizable Production of Label-Free Sensor Arrays.
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DOI：
10.1021/acs.analchem.5b01669
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发表时间：
2015-08-04
影响因子：
7.4
通讯作者：
Miller BL
中科院分区：
文献类型：
--
作者：
Lifson MA;Carter JA;Miller BL
将探针分子固定到表面是许多分析装置（包括标记和无标记微阵列）生产中的关键步骤。提高探针沉积密度和均匀性的新方法有可能显着提高检测的最终极限和再现性。过去，基于水凝胶的材料已被用来提供 3D 蛋白质友好的表面，用于沉积抗体和核酸。然而，这些方法在水凝胶支架聚合过程中容易受到变化的影响，并且为逐个抗体调整沉积参数提供了有限的机会。在这项工作中，开发了一种多功能水凝胶纳米颗粒沉积方法，用于生产无标记微阵列，并在抗体-抗原结合的背景下进行了测试。使用抗生物素蛋白/生物素系统将聚（N-异丙基丙烯酰胺）纳米颗粒（PNIPAM）与抗体缀合，并使用非接触式印刷系统沉积到表面上。干燥后，这些凝胶点形成高度< 10 nm 的均匀薄层。通过动态光散射、扫描电子显微镜和原子力显微镜对缀合物进行了表征。我们通过阵列成像反射法 (AIR)（一种无标记蛋白质微阵列方法）在肿瘤坏死因子-α (TNF-α) 检测中测试了这种格式。这种探针分子沉积方法通常可用于生产用于无标记检测的微阵列。
Probe molecule immobilization onto surfaces is a critical step in the production of many analytical devices, including labeled and label-free microarrays. New methods to increase the density and uniformity of probe deposition have the potential to significantly enhance the ultimate limits of detection and reproducibility. Hydrogel-based materials have been employed in the past to provide a 3D protein-friendly surface for deposition of antibodies and nucleic acids. However, these methods are susceptible to variation during polymerization of the hydrogel scaffold, and provide limited opportunities for tuning deposition parameters on an antibody-by-antibody basis. In this work, a versatile hydrogel nanoparticle deposition method was developed for the production of label-free microarrays, and tested in the context of antibody-antigen binding. Poly (N-isopropylacrylamide) nanoparticles (PNIPAM) were conjugated to antibodies using an avidin/biotin system and deposited onto surfaces using a noncontact printing system. After drying, these gel spots formed uniform and thin layers < 10 nm in height. The conjugates were characterized with dynamic light scattering, scanning electron microscopy, and atomic force microscopy. We tested this format in the context of tumor necrosis factor-alpha (TNF-α) detection via Arrayed Imaging Reflectometry (AIR), a label-free protein microarray method. This method of probe molecule deposition should be generally useful in the production of microarrays for label-free detection.