Simultaneous evaluation of multiple microarray surface chemistries through real-time interferometric imaging
Simultaneous evaluation of multiple microarray surface chemistries through real-time interferometric imaging
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DOI：
10.1007/s00216-019-02276-1
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发表时间：
2020-01-04
影响因子：
4.3
通讯作者：
Chiari, Marcella
中科院分区：
文献类型：
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作者：
Chiodi, Elisa;Sola, Laura;Chiari, Marcella
表面化学是微阵列生物传感器开发的一个重要方面。功能化表面的固定能力确实是结合反应最终产率的限制因素。在这项工作中，我们能够同时比较局部固定在不同聚合物上和同一固体支持物上的蛋白质配体的功能，从而展示了一种新的多重分析方法。我们的目标是在一次实验中研究一组反应性聚合物的固定效率以及所束缚分子的亲和力。通过在 Si/SiO2 芯片上点样许多反应性聚合物并随后立即将分子探针沉积在这些点上，证明了这一想法。作为概念验证，我们重点关注哪种聚合物可以更好地固定模型蛋白（α-乳清蛋白）和肽（LAC-1）。我们成功地证明该方案适用于蛋白质和肽，并且具有良好的效率。通过使用干涉反射成像传感器（IRIS）进行实时结合测量，局部功能化被证明与经典的平面涂层解决方案相当。最终结果凸显了该方法的多重功能：首先，它可以同时表征数十种聚合物。其次，它消除了与涂层表面相关的限制，即只有具有相同官能团的分子才能连接到相同的固体支持物上。通过应用该方案，可以同时研究多种类型的分子，并且可以单独优化每个探针的固定。
Surface chemistry is a crucial aspect for microarray modality biosensor development. The immobilization capability of the functionalized surface is indeed a limiting factor for the final yield of the binding reaction. In this work, we were able to simultaneously compare the functionality of protein ligands that were locally immobilized on different polymers, while on the same solid support, therefore demonstrating a new way of multiplexing. Our goal was to investigate, in a single experiment, both the immobilization efficiency of a group of reactive polymers and the resulting affinity of the tethered molecules. This idea was demonstrated by spotting many reactive polymers on a Si/SiO2 chip and depositing the molecular probes on the spots immediately after. As a proof of concept, we focused on which polymers would better immobilize a model protein (alpha-Lactalbumin) and a peptide (LAC-1). We successfully showed that this protocol is applicable to proteins and peptides with a good efficiency. By means of real-time binding measurements performed with the interferometric reflectance imaging sensor (IRIS), local functionalization proved to be comparable to the classical flat coating solution. The final outcome highlights the multiplexing power of this method: first, it allows to characterize dozens of polymers at once. Secondly, it removes the limitation, related to coated surfaces, that only molecules with the same functional groups can be tethered to the same solid support. By applying this protocol, many types of molecules can be studied simultaneously and immobilization for each probe can be individually optimized.