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通过包埋在凝胶网络中来固定酶。

Enzyme immobilization by entrapment within a gel network.

作者信息

Sassolas Audrey, Hayat Akhtar, Marty Jean-Louis

机构信息

IMAGES EA, University of Perpignan, Perpignan Cedex, France.

出版信息

Methods Mol Biol. 2013;1051:229-39. doi: 10.1007/978-1-62703-550-7_15.

Abstract

This chapter provides a detailed description of the three immobilization methods based on the biomolecules entrapment into polymer matrices. The poly (vinyl alcohol) bearing styrylpyridinium groups (PVA-SbQ), a soluble pre-polymer bearing photo-cross-linkable groups, has widely been used to entrap enzymes, and several bioassays based on this immobilization matrix have been reported. Similarly, immobilization of enzymes via sol-gel has been described in this chapter. Sol-gel process is based on the ability to form solid metal or semi-metal oxides via the aqueous process of hydrolytically labile precursors. Enzymes can also be entrapped in an agarose gel. Contrary to synthetic polymers such as polyacrylamide, this matrix is biocompatible, non-toxic, provides natural microenvironment to the enzyme and also gives sufficient accessibility to electrons to shuttle between the enzyme and the electrode. The entrapment strategies are easy-to-perform, and permit to deposit enzyme, mediators, and additives in the same sensing layer. Moreover, the activity of the enzyme is preserved during the immobilization process, as biological element is not modified. Biosensors based on physically entrapped enzymes are often characterized by increased operational and storage stability.

摘要

本章详细描述了基于生物分子包埋于聚合物基质中的三种固定化方法。带有苯乙烯基吡啶鎓基团的聚乙烯醇(PVA-SbQ),一种带有可光交联基团的可溶性预聚物,已被广泛用于包埋酶,并且基于这种固定化基质的几种生物测定方法已有报道。同样,本章也描述了通过溶胶-凝胶法固定酶的方法。溶胶-凝胶法基于通过水解不稳定前体的水相过程形成固体金属或半金属氧化物的能力。酶也可以被包埋在琼脂糖凝胶中。与聚丙烯酰胺等合成聚合物不同,这种基质具有生物相容性、无毒,为酶提供天然微环境,并且还能为电子在酶和电极之间穿梭提供足够的可及性。包埋策略易于实施,并且允许在同一传感层中沉积酶、媒介物和添加剂。此外,在固定化过程中酶的活性得以保留,因为生物元件未被修饰。基于物理包埋酶的生物传感器通常具有更高的操作稳定性和储存稳定性。

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