【作者】 杨清林；

【导师】 蔡绍皙；

【作者基本信息】 重庆大学 ， 生物医学工程， 2005， 硕士

【摘要】 具有生物膜结构和生物相容性的双分子层膜(BLMs)易于修饰,将适当的活性分子包埋进脂双分子层中应当能够赋予BLMs以相应的功能特征;胆固醇能调节BLMs的流动性和增加BLMs 的稳定性,所以胆固醇在BLMs 的含量对于基于有支撑的脂双分子层膜(supported BLM,s-BLM)的生物传感器十分重要。因此通过实验,研究了不同量的胆固醇对s-BLM 稳定性的影响,提出了一种新的形成有胆固醇修饰的s-BLM 的方法,并利用这种方法研究了一种生物膜电化学技术,基于表面稳定的由鸡蛋卵磷脂组成的BLMs,它对寡聚单链脱氧核糖核酸(ss-DNA),即胸腺嘧啶脱氧核糖核酸二十核苷(Oligo d(T20))具有良好的响应。本文就是在研究不同量的胆固醇修饰s-BLM 的基础之上,通过其对ss-DNA 响应的分析,希望可能构建一种基于整合有ss-DNA 的s-BLM 生物传感器装置。本文的主要研究工作和结论如下: (1)研究了不同量的胆固醇对s-BLM稳定性的影响: 本文利用循环伏安法(CV)对比研究了不同量的胆固醇修饰成膜,对膜的稳定性的影响;结果表明,胆固醇与磷脂的摩尔比为1 ∶1 时,s-BLM的稳定性最好。(2)研究了s-BLM对连接有四氯荧光素(tetrachloro-fluorescein,TET)的Oligo d(T20)的响应: 将连接有TET的ss-DNA加入电解质溶液中,观察s-BLM对连接有TET的Oligo d(T20)的响应,结果发现,s-BLM对连接有TET的Oligo d(T20)的响应在大约14分钟出现,首先峰电流突然升高,然后逐步下降,最后趋于平稳。(3)研究了s-BLM 对连接有嵌二萘(pyrene)的Oligo d(T20)的响应: 将连接有pyrene 的ss-DNA 加入电解质溶液中,观察s-BLM 对连接有pyrene的Oligo d(T20)的响应,结果发现,s-BLM 对连接有pyrene 的Oligo d(T20)的响应在大约9 分钟出现,首先峰电流突然升高,然后逐步下降,最后趋于平稳。从s-BLM 对连接有pyrene 和TET 的Oligo d(T20)的响应比较来看,前者的响应时间比后者要快,峰电流值的变化要大。另外,研究发现,ss-DNA 修饰s-BLM,形成整合有ss-DNA 的s-BLM,s-BLM稳定性显著提高。(4)构建基于s-BLM 的微阵列传感器模型: 提出一个基于s-BLM 的微型阵列生物传感器模型,将s-BLM 与微电子技术和微加工技术相结合,可以实现对多目标进行同时检测。更多还原

【Abstract】 Bilayer Lipid Membranes (BLMs), with the bilayer structure of biomembrance as well as good biological compatibility, is prone to be modified. BLMs can be endowed corresponding functions after embedding certain active molecules in BLMs. Cholesterol can adjust mobility of BLMs and enhance stability of BLMs. The proportion of cholesterol in BLMs plays important roles in biosensor based on supported BLMs. Thus, effects of different amount of cholesterol on stability of s-BLM were investigated. The dissertation advanced a novel formation method of s-BLM. By means of this new way we describes an electrochemical technique that exists the satisfying respond to single stranded deoxyribonucleic acids(ss-DNA) : thymidylic acid icosanucleotide(Oligo d(T20) ) based on surface stabilized s-BLM which is composed of egg phosphatidylcholine (PC). S-BLM modified with different amount of cholesterol and then their responses to ss-DNA were analyzed. We expected to establish biosensor system based on s-BLM integrated with ss-DNA. The main work and conclusions are as follows: (1) The stability influences of the different amount of cholesterol to s-BLM were studied. Influences of cholesterol to s-BLM were studied by comparing s-BLM modified with different amount cholesterol with Cyclic Voltammetry. The results indicated that s-BLM got the best stability of when the mol ratio of cholesterol and egg phosphatidylcholine is 1:1 in the BLM. (2) The responses of s-BLM to ss-DNA linked with tetrachloro-fluorescein (TET) were investigated. The responses of s-BLM to ss-DNA were observed in electrolyte solution with adding ss-DNA modified with TET. As a result,the response of s-BLM to ss-DNA turned up after 14 minutes. The peak current value appeared suddenly at first and then decreased gradually with leveled off in the end. (3) The responses of s-BLM to ss-DNA linked with pyrene were studied. Ss-DNA modified with pyrene was put into electrolyte solution and the responses of s-BLM to ss-DNA linked with pyrene were studied. The results showed the similar results to ss-DNA linked with TET. There were responses after 9 minutes. At first the peak current value suddenly come and then dropped gradually and finally with leveled off. Compared the responses of s-BLM to ss-DNA linked with TET to ss-DNA linked with pyrene, the response time of the former is shorter than the latter and the change extend of peak current value of the former is larger. In addition,the stability of s-BLM was improved greatly after modifying with ss-DNA. (4) The microarray biosensor based on s-BLM was built. The microarray biosensor model based on s-BLM was put forward. The microarray biosensor can examine multi-objects at the same time with combing micro-electron with micro-machining technology.更多还原