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互穿聚合物网络捕捉可供酶测定用的 FRET 敏感聚合物。

Interpenetrating Polymer Network Capturing FRET-Sensitive Polymers Available for an Enzyme Assay.

机构信息

Area for Molecular Function, Division of Material Science, Graduate School of Science and Engineering, Saitama University, Sakura, Saitama 338-8570, Japan.

Medical Innovation Research Unit (MiU), Advanced Institute of Innovative Technology (AIIT), Saitama University, Sakura, Saitama 338-8570, Japan.

出版信息

Biomacromolecules. 2024 Aug 12;25(8):5222-5232. doi: 10.1021/acs.biomac.4c00622. Epub 2024 Aug 1.

Abstract

Fluorogenic glycomonomers have been used for biological evaluations, and water-soluble and Förster resonance energy transfer (FRET)-sensitive glycopolymers have also been reported. A FRET-sensitive polymer was conveniently prepared from a fluorogenic donor monomer and a fluorogenic acceptor monomer by means of simple radical polymerization in high yield. Continuous fluorospectroscopic monitoring of the polymer in the presence of an enzyme was performed, and the results showed the possible application of the FRET-sensitive glycopolymer for practical use. In addition to the use of aqueous solution phase, the water-soluble and FRET-sensitive glycopolymer was completely captured into an interpenetrating polymer network (IPN) by means of radical polymerization with a combination of acrylamide and bis-acrylamide as used for the cross-linking reagent system. The IPN including the FRET-sensitive glycopolymer was allowed to react with amylases in an aqueous buffer solution at 37 °C, and the enzymatic reaction was continuously and conveniently monitored by means of fluorometric spectroscopy.

摘要

荧光糖单体已被用于生物学评估,并且还报道了水溶性和Förster 共振能量转移(FRET)敏感糖聚合物。通过简单的自由基聚合,可以高产率地将荧光供体单体和荧光受体单体方便地制备成 FRET 敏感聚合物。在存在酶的情况下,通过连续荧光光谱监测聚合物,结果表明 FRET 敏感糖聚合物可能适用于实际用途。除了使用水相之外,水溶性和 FRET 敏感糖聚合物还通过使用丙烯酰胺和双丙烯酰胺作为交联试剂体系的组合的自由基聚合完全捕获到互穿聚合物网络(IPN)中。包含 FRET 敏感糖聚合物的 IPN 在 37°C 的水性缓冲溶液中与淀粉酶反应,并且通过荧光光谱法连续且方便地监测酶反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fc9b/11323839/b942306ac003/bm4c00622_0001.jpg

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