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用于葡萄糖微生物传感器的新型 SU-8 微加工膜中的酶固定。

A novel enzyme entrapment in SU-8 microfabricated films for glucose micro-biosensors.

机构信息

Cranfield University, Cranfield Health, Cranfield, Bedfordshire MK43 0AL, UK.

出版信息

Biosens Bioelectron. 2010 Dec 15;26(4):1582-7. doi: 10.1016/j.bios.2010.07.117. Epub 2010 Aug 5.

DOI:10.1016/j.bios.2010.07.117
PMID:20732802
Abstract

The present work investigates the utilisation of the widely used SU-8 photoresist as an immobilisation matrix for glucose oxidase (GOx) for the development of glucose micro-biosensors. The strong advantage of the proposed approach is the simultaneous enzyme entrapment during the microfabrication process within a single step, which is of high importance for the simplification of the BioMEMS procedures. Successful encapsulation of the enzyme GOx in "customised" SU-8 microfabricated structures was achieved through optimisation of the one-step microfabrication process. Although the process involved contact with organic solvents, UV-light exposure, heating for pre- and post-bake and enzyme entrapment in a hard and rigid epoxy resin matrix, the enzyme retained its activity after encapsulation in SU-8. Measurements of the immobilised enzyme's activity inside the SU-8 matrix were carried out using amperometric detection of hydrogen peroxide in a 3-electrode setup. Films without enzyme showed negligible variation in current upon the addition of glucose, as opposed to films with encapsulated enzyme which showed a very clear increase in current. Experiments using films of increased thickness or enzyme concentration, showed a higher response, thus proving that the enzyme remained active not only on the film's surface, but also inside the matrix as well. The proposed enzyme immobilisation in SU-8 films opens up new possibilities for combining BioMEMS with biosensors and organic electronics.

摘要

本工作研究了广泛使用的 SU-8 光刻胶作为葡萄糖氧化酶 (GOx) 的固定基质,用于开发葡萄糖微生物传感器。所提出的方法的一个主要优势是可以在单个步骤中同时进行酶包埋,这对于简化 BioMEMS 工艺非常重要。通过优化一步微加工工艺,成功地将酶 GOx 包埋在“定制”的 SU-8 微加工结构中。尽管该过程涉及与有机溶剂接触、UV 光暴露、预烘和后烘加热以及在坚硬刚性环氧树脂基质中包埋酶,但在 SU-8 中包埋后,酶仍保持其活性。使用三电极设置通过安培检测法在 SU-8 基质内测量固定化酶的活性。与包埋有酶的薄膜相比,不含酶的薄膜在添加葡萄糖时电流几乎没有变化,而包埋有酶的薄膜电流明显增加。使用增加厚度或酶浓度的薄膜进行的实验显示出更高的响应,这证明酶不仅在薄膜表面保持活性,而且在基质内部也保持活性。在 SU-8 薄膜中固定化酶为将 BioMEMS 与生物传感器和有机电子学结合起来开辟了新的可能性。

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