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琥珀酰化 Jeffamine ED-2003 涂层聚碳酸酯芯片,用于低成本分析微阵列。

Succinylated Jeffamine ED-2003 coated polycarbonate chips for low-cost analytical microarrays.

机构信息

Institute of Hydrochemistry, Chair of Analytical Chemistry and Water Chemistry, Technical University of Munich, Marchioninistrasse 17, 81377, Munich, Germany.

出版信息

Anal Bioanal Chem. 2019 Apr;411(10):1943-1955. doi: 10.1007/s00216-019-01594-8. Epub 2019 Feb 11.

Abstract

Analytical microarrays feature great capabilities for simultaneous detection and quantification of multiple analytes in a single measurement. In this work, we present a rapid and simple method for bulk preparation of microarrays on polycarbonate sheets. Succinylated Jeffamine® ED-2003 was screen printed on polycarbonate sheets to create a polyfunctional shielding layer by baking at 100 °C. After microdispension of capture probes (antibodies, oligonucleotides, or small molecules) in a microarray format, chips were assembled with a flow cell from double-sided tape. It was shown that the shielding layer was firmly coated and suppressed unspecific binding of proteins. Universal applicability was demonstrated by transferring established flow-based chemiluminescence microarray measurement principles from glass slides to polycarbonate chips without loss of analytical performance. Higher chemiluminescence signals could be generated by performing heterogeneous asymmetric recombinase polymerase amplification on polycarbonate chips. Similar results could be shown for sandwich microarray immunoassays. Beyond that, lower inter- and intra-assay variances could be measured for the analysis of Legionella pneumophila Serogroup 1, strain Bellingham-1. Even surface regeneration of indirect competitive immunoassays was possible, achieving a limit of detection of 0.35 ng L for enrofloxacin with polycarbonate microarray chips. Succinylated Jeffamine ED-2003 coated polycarbonate chips have great potential to replace microtiter plates by flow-based chemiluminescence microarrays for rapid analysis. Therefore, it helps analytical microarrays to advance into routine analysis and diagnostics. Graphical abstract ᅟ.

摘要

分析微阵列具有在单次测量中同时检测和定量多种分析物的强大功能。在这项工作中,我们提出了一种在聚碳酸酯片上快速简便地制备微阵列的方法。将琥珀酰化 Jeffamine® ED-2003 通过在 100°C 下烘烤而被印刷到聚碳酸酯片上,以形成多功能屏蔽层。在微阵列格式中分散捕获探针(抗体、寡核苷酸或小分子)之后,用双面胶带制成的流动池将芯片组装在一起。结果表明,该屏蔽层被牢固地涂覆,并抑制了蛋白质的非特异性结合。通过将基于流动的化学发光微阵列测量原理从载玻片转移到聚碳酸酯芯片上,而不损失分析性能,证明了其普遍适用性。通过在聚碳酸酯芯片上进行异质不对称重组酶聚合酶扩增,可以产生更高的化学发光信号。夹心微阵列免疫测定也可以得到类似的结果。除此之外,对于军团菌血清群 1 菌株 Bellingham-1 的分析,还可以测量到较低的组内和组间变异。甚至可以实现间接竞争免疫测定的表面再生,用聚碳酸酯微阵列芯片实现恩诺沙星的检测限为 0.35ng/L。用琥珀酰化 Jeffamine ED-2003 涂覆的聚碳酸酯芯片具有通过基于流动的化学发光微阵列快速分析来替代微滴定板的巨大潜力。因此,它有助于分析微阵列进入常规分析和诊断。

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