血红蛋白仿生膜的电化学传感研究
发布时间：2018-10-21 18:30
【摘要】： 血红蛋白(Hemoglobin, Hb)是脊椎动物红细胞内的呼吸蛋白,由珠蛋白和血红素组成,是血液中分子氧的载体。研究血红蛋白的电化学行为,不仅可以探索生物大分子之间的长程电子传递的机理,进一步认识生物大分子的结构与功能之间的关系,而且有助于我们了解生命体系内的物质代谢和能量交换的过程及生命过程的氧化还原机理,从而更有效地开发和研制具有高催化性能的电化学生物传感器。 本论文以实现血红蛋白的直接电子传递为目的,分别选取具有良好生物相容性的碳纳米材料、高分子材料等仿生材料作为血红蛋白的固定材料,并采用适当的方法将血红蛋白固定于电极表面,基于这些材料与血红蛋白之间的特殊相互作用,实现了血红蛋白与基底电极之间的直接电子传递。这些固定材料不但为血红蛋白保持自然结构和生物活性提供了适宜的微环境,而且显著提高了血红蛋白与电极间的电子传递速率。基于此制备的化学修饰电极能够催化还原过氧化氢(H_2O_2)、一氧化氮(NO)等小分子。本论文的主要工作有: (1)首次将乙炔黑与离子液体1-丁基-3-甲基咪唑六氟磷酸盐(BMIMPF_6)组合制得复合膜,并成功将Hb固定在电极表面,为Hb的电化学研究建立了新的平台。用紫外-可见光谱法和电化学交流阻抗对Hb/AB-BMIMPF_6复合膜进行了表征,结果表明Hb在AB-BMIMPF_6复合膜中保持其生物活性,同时该膜促进了Hb与电极间电子的传递。该修饰电极对H_2O_2、NO的还原具有良好的催化作用。 (2)采用一种新型合成的高分子材料-纤维素季铵盐,利用静电吸附作用,采用层层自组装法将荷负电的血红蛋白包埋于荷正电的纤维素季铵盐内,并固定在电极表面,从而实现了血红蛋白的直接电化学。研究表明,固定在纤维素季铵盐膜内的血红蛋白没有变性,能催化还原H_2O_2。基于此,我们构建了一种无媒介体的新型过氧化氢传感器。 (3)结合亚甲基蓝和纳米活性炭微球的优异性能,制备出一种复合膜,并成功实现Hb的包埋固定。Hb在此复合膜内表现出优良的直接电化学行为。原子力显微镜、电化学交流阻抗、紫外-可见光谱研究表明,该复合膜与Hb之间存在着静电作用,它改变Hb的结构取向,因而更利于电子传递。固定于亚甲基蓝-纳米活性炭微球复合膜内的Hb也表现出了良好的生物催化活性,能催化还原H_2O_2。 (4)采用β-环糊精和离子液体1-丁基-3-甲基咪唑四氟硼酸盐(BMIMBF_4)制得修饰膜,利用它们优异的生物相容性包埋固定血红蛋白,实现了血红蛋白在电极表面的直接电子传递。紫外-可见光谱实验表明,固定于β-环糊精-BMIMBF_4复合膜内的血红蛋白仍然能维持其原始构象和生物活性。该复合膜修饰的玻碳电极能有效地催化H_2O_2还原,同时响应快、稳定性高。
[Abstract]:Hemoglobin (Hemoglobin, Hb) is a respiratory protein in erythrocytes of vertebrates. It is composed of globin and heme, and is the carrier of molecular oxygen in blood. The study of the electrochemical behavior of hemoglobin can not only explore the mechanism of long-range electron transport among biomolecules, but also further understand the relationship between the structure and function of biological macromolecules. It also helps us to understand the process of substance metabolism and energy exchange and the mechanism of redox in the life process, so that the electrochemical biosensor with high catalytic performance can be developed and developed more effectively. In order to realize the direct electron transfer of hemoglobin, carbon nanomaterials with good biocompatibility and biomimetic materials such as polymer materials were selected as the immobilized materials of hemoglobin. The direct electron transfer between hemoglobin and substrate electrode was realized based on the special interaction between hemoglobin and hemoglobin. These immobilization materials not only provide a suitable microenvironment for hemoglobin to maintain its natural structure and biological activity, but also significantly increase the electron transfer rate between hemoglobin and electrode. The chemically modified electrode can catalyze the reduction of hydrogen peroxide (H_2O_2), nitric oxide (NO) and other small molecules. The main work of this thesis is as follows: (1) the composite membrane was prepared by the combination of acetylene black and ionic liquid 1 Ding Ji 3 methyl imidazolium hexafluorophosphate (BMIMPF_6) for the first time, and Hb was successfully immobilized on the electrode surface. A new platform for electrochemical research of Hb is established. The Hb/AB-BMIMPF_6 composite film was characterized by UV-Vis spectroscopy and electrochemical impedance spectroscopy. The results showed that Hb kept its biological activity in the AB-BMIMPF_6 composite film and promoted the electron transfer between the Hb and the electrode. The modified electrode has a good catalytic effect on the reduction of H _ 2O _ 2no. (2) A new synthetic polymer material, cellulose quaternary ammonium salt, is used for electrostatic adsorption. The hemoglobin with negative charge was encapsulated in the positively charged quaternary ammonium cellulose salt and immobilized on the surface of the electrode by means of layer by layer self-assembly method, thus realizing the direct electrochemistry of hemoglobin. The results showed that hemoglobin immobilized in cellulose quaternary ammonium salt film had no denaturation and could catalyze the reduction of H _ 2O _ 2. Based on this, we constructed a novel hydrogen peroxide sensor without medium. (3) A composite membrane was prepared by combining the excellent properties of methylene blue and nano-activated carbon microspheres. The encapsulation and fixation of Hb were successfully realized. Hb showed excellent direct electrochemical behavior in the composite film. Atomic force microscopy (AFM) electrochemical impedance spectroscopy (EIS) and UV-Vis spectroscopy show that there is electrostatic interaction between the composite film and Hb which changes the structural orientation of Hb and is more favorable for electron transfer. The Hb immobilized in the composite membrane of methylene blue and nano-activated carbon also showed good biocatalytic activity. The modified membrane was prepared from 尾 -cyclodextrin and ionic liquid 1-Ding Ji -3-methyl imidazolium tetrafluoroborate (BMIMBF_4), and hemoglobin was immobilized by their excellent biocompatibility. The direct electron transfer of hemoglobin on the electrode surface was realized. UV-Vis spectra showed that hemoglobin immobilized in 尾 -cyclodextrin-BMIMBF_4 composite membrane could still maintain its original conformation and biological activity. The modified glassy carbon electrode can effectively catalyze the reduction of H_2O_2 with fast response and high stability.
【学位授予单位】：湘潭大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R341
本文编号：2285983
[Abstract]:Hemoglobin (Hemoglobin, Hb) is a respiratory protein in erythrocytes of vertebrates. It is composed of globin and heme, and is the carrier of molecular oxygen in blood. The study of the electrochemical behavior of hemoglobin can not only explore the mechanism of long-range electron transport among biomolecules, but also further understand the relationship between the structure and function of biological macromolecules. It also helps us to understand the process of substance metabolism and energy exchange and the mechanism of redox in the life process, so that the electrochemical biosensor with high catalytic performance can be developed and developed more effectively. In order to realize the direct electron transfer of hemoglobin, carbon nanomaterials with good biocompatibility and biomimetic materials such as polymer materials were selected as the immobilized materials of hemoglobin. The direct electron transfer between hemoglobin and substrate electrode was realized based on the special interaction between hemoglobin and hemoglobin. These immobilization materials not only provide a suitable microenvironment for hemoglobin to maintain its natural structure and biological activity, but also significantly increase the electron transfer rate between hemoglobin and electrode. The chemically modified electrode can catalyze the reduction of hydrogen peroxide (H_2O_2), nitric oxide (NO) and other small molecules. The main work of this thesis is as follows: (1) the composite membrane was prepared by the combination of acetylene black and ionic liquid 1 Ding Ji 3 methyl imidazolium hexafluorophosphate (BMIMPF_6) for the first time, and Hb was successfully immobilized on the electrode surface. A new platform for electrochemical research of Hb is established. The Hb/AB-BMIMPF_6 composite film was characterized by UV-Vis spectroscopy and electrochemical impedance spectroscopy. The results showed that Hb kept its biological activity in the AB-BMIMPF_6 composite film and promoted the electron transfer between the Hb and the electrode. The modified electrode has a good catalytic effect on the reduction of H _ 2O _ 2no. (2) A new synthetic polymer material, cellulose quaternary ammonium salt, is used for electrostatic adsorption. The hemoglobin with negative charge was encapsulated in the positively charged quaternary ammonium cellulose salt and immobilized on the surface of the electrode by means of layer by layer self-assembly method, thus realizing the direct electrochemistry of hemoglobin. The results showed that hemoglobin immobilized in cellulose quaternary ammonium salt film had no denaturation and could catalyze the reduction of H _ 2O _ 2. Based on this, we constructed a novel hydrogen peroxide sensor without medium. (3) A composite membrane was prepared by combining the excellent properties of methylene blue and nano-activated carbon microspheres. The encapsulation and fixation of Hb were successfully realized. Hb showed excellent direct electrochemical behavior in the composite film. Atomic force microscopy (AFM) electrochemical impedance spectroscopy (EIS) and UV-Vis spectroscopy show that there is electrostatic interaction between the composite film and Hb which changes the structural orientation of Hb and is more favorable for electron transfer. The Hb immobilized in the composite membrane of methylene blue and nano-activated carbon also showed good biocatalytic activity. The modified membrane was prepared from 尾 -cyclodextrin and ionic liquid 1-Ding Ji -3-methyl imidazolium tetrafluoroborate (BMIMBF_4), and hemoglobin was immobilized by their excellent biocompatibility. The direct electron transfer of hemoglobin on the electrode surface was realized. UV-Vis spectra showed that hemoglobin immobilized in 尾 -cyclodextrin-BMIMBF_4 composite membrane could still maintain its original conformation and biological activity. The modified glassy carbon electrode can effectively catalyze the reduction of H_2O_2 with fast response and high stability.
【学位授予单位】：湘潭大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R341
【参考文献】
相关期刊论文 前8条
1 张玉忠;苏邵;郝和群;蔡跃娟;倪永红;;血红蛋白在DHP/PAM-CdS膜修饰金电极上的直接电化学及其与利巴韦林相互作用的研究[J];安徽师范大学学报(自然科学版);2007年03期
2 马豫峰,蔡继业,杨培慧,陈勇;壳聚糖自组装复合修饰血红蛋白的研究[J];高分子材料科学与工程;2005年01期
3 董文宾,胡献丽,郑丹,杨兆艳;生物传感器在水质分析监测中的应用[J];工业水处理;2005年03期
4 穆冬燕;麦芽糊精修饰测酚酪氨酸酶膜电极的研制[J];环境工程;2004年05期
5 李平,刘梅川,张成林,程欲晓,张莉,金利通;聚乙烯吡咯烷酮/硫化镉量子点修饰电极的制备及其对血红蛋白的测定研究[J];化学学报;2005年12期
6 武五爱;郭满栋;尹志芬;尉景瑞;李婧芳;李君;张宇;;血红蛋白在纳米孔径氧化铝膜修饰电极上的直接电化学[J];化学学报;2009年08期
7 ;Direct electrochemistry and enzymatic activity of hemoglobin in positively charged colloid Au nanoparticles and hemoglobin layer-by-layer self-assembly films[J];Science in China(Series B:Chemistry);2007年05期
8 王新莹;蔡文怡;姜立萍;朱俊杰;;银-磷酸钛纳米复合膜的构建及其电化学生物传感[J];无机化学学报;2008年12期
，本文编号：2285983
本文链接：https://www.wllwen.com/yixuelunwen/shiyanyixue/2285983.html
最近更新
教材专著
