摘要：蛋白质硝基化损伤可能导致蛋白质结构功能改变，诱导细胞损伤和凋亡，最终 引发疾病。硝基自由基(NO•)对蛋白质的损伤检测对于研究其致病机制具有重要的科学意 义。本论文拟结合纳米复合材料比表面及高、导电性优异、生物相容性好的优势，一步法 电还原构建灵敏度高、制备过程简单的还原态石墨烯(rGO)-纳米金(AuNPs)-氯化高铁血红 素(Hemin)修饰的电化学生物传感器( rGO/AuNPs /GCE)，利用 NaNO2/Hemin/H2O2 体系对 吸附牛血清白蛋白(BSA)产生硝基化损伤，利用差分脉冲伏安法(DPV)对硝基化 BSA 进行 了灵敏检测，并优化了循环伏安法沉积的圈数，沉积液的浓度比例等实验条件。实验结果 表明：使用 rGO/AuNPs /GCE 电极，在 0.1mol/L pH 7.0 的磷酸盐缓冲溶液(PBS)中，峰电 流与硝基化 BSA 在浓度范围 1×10-7~1×10-5mol/L 内呈现出良好的线性关系，检出限为 1.926×10-6mol/L (3S/N) 。66329

毕业论文关键词: 蛋白质硝基化；电化学传感器；石墨烯；纳米金

Detection of Protein Nitrification Damage Based on Electrochemical Biosensor

Abstract：Nitrolysis of proteins may lead to changes in protein structure and function, induce cell damage and apoptosis, and ultimately cause disease. Nitrocellulose (NO) attack on protein is of great scientific significance in the study of its pathogenic mechanism. In this paper, we propose a new method to reconstruct the gravimetric gravimetric (rGO) - gold nanoparticles (AuNPs) - based on the high surface area, high conductivity, excellent biocompatibility and good biocompatibility of the nanocomposites. The electrochemical biosensor (rGO/AuNPs/GCE) modified by Hemin was used to generate nitration damage by adsorbent bovine serum albumin (BSA) using NaNO2/Hemin/H2O2 system. Using differential pulse voltammetry (DPV) was used to detect nitrolylated BSA, and the experimental conditions  such

as the number of turns of cyclic voltammetry deposition and the concentration ratio of sediment were optimized. The results showed that the peak current was in the range of 1 × 10-7 ~ 1 × 10-5mol in the concentration range of nitric oxide (BSA) in 0.1mol / L phosphate buffer solution (PBS) at pH 7.0 with rGO/AuNPs/GCE electrode / L showed a good linear relationship, the detection limit of 1.926 × 10-6mol / L (3S / N).

Key words: Protein nitrocellulose；Electrochemical sensor；Graphene；Gold Nanopartic

目录

1 绪论 1

1.1  蛋白质 1

1.1.1  蛋白质翻译后修饰 1

1.1.2  蛋白质硝基化 3

1.1.3  牛血清白蛋白 5

1.2  生物传感器 5

1.2.1  生物传感器的分类 6

1.2.2  电化学生物传感器 7

1.3  纳米材料 7

1.3.1  氧化石墨烯 7

1.3.2  纳米金颗粒 9

1.5  基于纳米材料的电化学生物传感器 9

1.6  本论文的研究与思路 11

2 本论文的技术路线与流程 12

3 实验方法与材料 12

3.1  实验材料 12

3.1.1  主要试剂 12

3.1.2 主要仪器 13

3.2  实验步骤