基于无机纳米片构筑第三代电化学生物传感器的研究
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摘要
基于酶在电极上直接电子转移的第三代电化学生物传感器是当前生物传感器的研究热点,而纳米材料所具有的独特物理化学性质为第三代电化学生物传感器的构建提供了一条崭新的途径。纳米片是一类新型的纳米材料,它具有各向异性、聚电解质等性质,可形成稳定高浓度的胶体溶液。本论文采用纳米片(磷酸锆纳米片、水滑石纳米片和氧化钛纳米片)固定肌红蛋白(Mb)或葡萄糖氧化酶(GOD),实现酶的直接电化学,构筑第三代电化学生物传感器。研究内容包括以下三部分:
1.采用磷酸锆纳米片(ZrPNS)作为固定Mb的载体,固定后的Mb活性中心没有泄漏。利用ZrPNS固定Mb修饰玻碳电极,电化学研究发现,ZrPNS促进了Mb与电极间快速有效的直接电子转移。修饰电极具有良好的热稳定性,对底物H_2O_2、O_2、NaNO_2和三氯乙酸具有较好的催化行为。同时,修饰电极也具备较好的抗干扰性、重现性、操作稳定性和储存稳定性。
2.采用水滑石纳米片(LDHNS)作为固定GOD的载体,LDHNS固定的GOD保持了良好的生物活性。利用LDHNS固定GOD修饰玻碳电极,电化学研究发现,LDHNS能够显著促进GOD的直接电化学反应,而对比实验发现,剥层前体层状水滑石固定的GOD在电极上仅能实现十分微弱的直接电化学行为。
3.采用氧化钛纳米片(TNS)利用层层组装法固定Mb,TNS固定化的Mb仍保持着良好的生物活性。在玻碳电极表面构筑TNS与Mb的层层组装膜,膜中的Mb实现了有效稳定的直接电子转移。层层组装膜中的Mb对底物O_2和H_2O_2具有良好的催化效应,催化效率较高。修饰电极具有良好的重现性和储存稳定性。
Much attention has been devoted to the development of the third-generation electrochemical biosensor based on direct electrochemistry of enzymes, and nanomaterials provide a new way to construct the third-generation biosensor. Nanosheets, a novel class of nanoscale materials, possess anisotropic morphology, polyelectrolytic and colloidal nature. It is significative that nanosheets are used to improve the direct electrochemistry between enzymes and electrodes.
In this thesis, the nanosheets (zirconium phosphate nanosheets, layered double hydroxide nanosheets and titania nanosheets) were used to immobilize myoglobin (Mb) or glucose oxidase (GOD) and investigate the electrochemical properties. The main contents of the thesis are given as follows:
1. Zirconium phosphate nanosheets (ZrPNS) were firstly applied to the immobilization of Mb as a support matrix. Mb almost retained bioactivity in the film. In ZrPNS film on the glassy carbon electrode, Mb realized its direct electrochemistry and displayed good thermal stability. The catalytic abilities of the protein toward the reduction of H_2O_2, O_2, NaNO_2 and trichloroacetic acid were also studied and a third-generation biosensor was subsequently fabricated. The biosensor also exhibited acceptable anti-interferant ability, reproducibility and stability.
2. Layered double hydroxide nanosheets (LDHNS) were firstly applied to the immobilization of GOD as a support matrix. GOD almost retained bioactivity in the film. The electrochemical results revealed that LDHNS could improve greatly the direct electrochemistry of GOD, while layered double hydroxide nanoparticles (LDHNP) can only achieve the direct electrochemistry of GOD in a very small extent.
3. Titania nanosheets (TNS) were applied to the immobilization of Mb via layer-by-layer (LBL) technology. Mb retained good bioactivity in the LBL film and realized effective direct electrochemistry behavior. The biosensor displayed high catalytic efficiency towards O_2 and H_2O_2, and possessed good reproducibility and stability.
1.采用磷酸锆纳米片(ZrPNS)作为固定Mb的载体,固定后的Mb活性中心没有泄漏。利用ZrPNS固定Mb修饰玻碳电极,电化学研究发现,ZrPNS促进了Mb与电极间快速有效的直接电子转移。修饰电极具有良好的热稳定性,对底物H_2O_2、O_2、NaNO_2和三氯乙酸具有较好的催化行为。同时,修饰电极也具备较好的抗干扰性、重现性、操作稳定性和储存稳定性。
2.采用水滑石纳米片(LDHNS)作为固定GOD的载体,LDHNS固定的GOD保持了良好的生物活性。利用LDHNS固定GOD修饰玻碳电极,电化学研究发现,LDHNS能够显著促进GOD的直接电化学反应,而对比实验发现,剥层前体层状水滑石固定的GOD在电极上仅能实现十分微弱的直接电化学行为。
3.采用氧化钛纳米片(TNS)利用层层组装法固定Mb,TNS固定化的Mb仍保持着良好的生物活性。在玻碳电极表面构筑TNS与Mb的层层组装膜,膜中的Mb实现了有效稳定的直接电子转移。层层组装膜中的Mb对底物O_2和H_2O_2具有良好的催化效应,催化效率较高。修饰电极具有良好的重现性和储存稳定性。
Much attention has been devoted to the development of the third-generation electrochemical biosensor based on direct electrochemistry of enzymes, and nanomaterials provide a new way to construct the third-generation biosensor. Nanosheets, a novel class of nanoscale materials, possess anisotropic morphology, polyelectrolytic and colloidal nature. It is significative that nanosheets are used to improve the direct electrochemistry between enzymes and electrodes.
In this thesis, the nanosheets (zirconium phosphate nanosheets, layered double hydroxide nanosheets and titania nanosheets) were used to immobilize myoglobin (Mb) or glucose oxidase (GOD) and investigate the electrochemical properties. The main contents of the thesis are given as follows:
1. Zirconium phosphate nanosheets (ZrPNS) were firstly applied to the immobilization of Mb as a support matrix. Mb almost retained bioactivity in the film. In ZrPNS film on the glassy carbon electrode, Mb realized its direct electrochemistry and displayed good thermal stability. The catalytic abilities of the protein toward the reduction of H_2O_2, O_2, NaNO_2 and trichloroacetic acid were also studied and a third-generation biosensor was subsequently fabricated. The biosensor also exhibited acceptable anti-interferant ability, reproducibility and stability.
2. Layered double hydroxide nanosheets (LDHNS) were firstly applied to the immobilization of GOD as a support matrix. GOD almost retained bioactivity in the film. The electrochemical results revealed that LDHNS could improve greatly the direct electrochemistry of GOD, while layered double hydroxide nanoparticles (LDHNP) can only achieve the direct electrochemistry of GOD in a very small extent.
3. Titania nanosheets (TNS) were applied to the immobilization of Mb via layer-by-layer (LBL) technology. Mb retained good bioactivity in the LBL film and realized effective direct electrochemistry behavior. The biosensor displayed high catalytic efficiency towards O_2 and H_2O_2, and possessed good reproducibility and stability.
引文
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