本文選題：電化學(xué)免疫傳感器 + 腫瘤標(biāo)志物��； 參考：《西南大學(xué)》2015年碩士論文

【摘要】：在醫(yī)學(xué)免疫分析中,發(fā)展一些快速、靈敏和準(zhǔn)確的檢測(cè)方法是科研工作者研究的重點(diǎn)、熱點(diǎn)問(wèn)題,這在臨床分析中對(duì)于疾病的診斷具有非常重要的意義。電化學(xué)免疫傳感器是一種將電化學(xué)技術(shù)、傳感器技術(shù)和免疫分析技術(shù)三者相結(jié)合而發(fā)展來(lái)的,集靈敏度高、選擇性好、操作簡(jiǎn)便等優(yōu)點(diǎn)于一身的分析技術(shù)。近幾十年來(lái),發(fā)展新型功能化的納米材料、探索生物大分子的固定化技術(shù)和構(gòu)建信號(hào)增強(qiáng)技術(shù)等用于電化學(xué)免疫傳感器檢測(cè)生物分子的研究受到了廣泛的關(guān)注。本文主要從功能化納米材料的制備,傳感器仿生界面的構(gòu)建,信號(hào)放大技術(shù)的創(chuàng)新等方面來(lái)進(jìn)行研究,并制備了一系列高靈敏的電化學(xué)免疫傳感器。主要研究?jī)?nèi)容如下：1.基于納米金功能化的多壁碳納米管復(fù)合材料標(biāo)記抗體的電化學(xué)免疫傳感器的研究本研究工作構(gòu)建了一種基于納米金/(3-巰丙基)三甲氧基硅烷(Au/MPTS)修飾傳感器界面和納米金功能化的多壁碳納米管復(fù)合材料標(biāo)記生物分子用于信號(hào)探針的夾心型電化學(xué)免疫傳感器,用于肝癌腫瘤標(biāo)志物甲胎蛋白(AFP)的檢測(cè)。首先,納米金通過(guò)PDDA共價(jià)交聯(lián)在多壁碳納米管表面制備得到了納米金功能化的碳納米材料,接著進(jìn)一步把電活性物質(zhì)硫堇(Thi)標(biāo)記的抗體生物分子和辣根過(guò)氧化物酶(HRP)固定在其表面,制備得到了納米生物探針。納米金功能化的多壁碳納米管固載了大量的生物分子和電活性物質(zhì),從而可以有效的提高免疫傳感器的靈敏度。該納米生物探針不僅能識(shí)別目標(biāo)分析物,而且在HRP對(duì)底物H2O2的催化作用下,可以進(jìn)一步放大電化學(xué)反應(yīng)信號(hào)。另外,Au/MPTS修飾傳感器界面不僅可以用來(lái)固載生物分子、保持其生物活性,還能進(jìn)一步促進(jìn)電子的傳遞。采用夾心免疫反應(yīng)模式,實(shí)現(xiàn)對(duì)AFP的定量檢測(cè),其線性范圍為0.01-50.0 ng·mL-1,檢測(cè)下限為3 pg·mL-1該傳感器具有良好的選擇性、穩(wěn)定性和重現(xiàn)性。2.基于雙酶催化沉淀技術(shù)和碳納米角構(gòu)建的信號(hào)增強(qiáng)型電化學(xué)免疫傳感器的研究在本實(shí)驗(yàn)中,結(jié)合碳納米角標(biāo)記和雙酶催化產(chǎn)物沉淀法構(gòu)建了一種新穎的電化學(xué)免疫傳感器,實(shí)現(xiàn)了對(duì)AFP的高靈敏檢測(cè)。首先,制備金-石墨稀復(fù)合納米材料(Au-Gra)用于修飾玻碳電極,不僅能促進(jìn)氧化還原探針與電極表面的電子傳遞,也能增加電極的有效面積提高AFP抗體的固載量。其次,將功能化處理后的碳納米角用于標(biāo)記抗體和固載葡萄糖氧化酶(GOD)和HRP得到生物納米探針,碳納米角較大的比表面積增加了抗體和雙酶的固載量。實(shí)驗(yàn)采用夾心免疫反應(yīng)模式,利用4-萘酚作為沉淀底物,該探針不僅能識(shí)別目標(biāo)分析物,還能通過(guò)雙酶催化沉淀實(shí)現(xiàn)信號(hào)放大,提高了免疫傳感器的靈敏度,從而實(shí)現(xiàn)對(duì)腫瘤標(biāo)志物的高靈敏檢測(cè)。免疫分析過(guò)程采用電化學(xué)交流阻抗和循環(huán)伏安法進(jìn)行表征。免疫傳感器對(duì)目標(biāo)分析物AFP的線性響應(yīng)范圍為0.001-60 ng·mL-1,檢測(cè)下限為0.33pg-mL-13.基于核-殼結(jié)構(gòu)Au@Pd雙金屬納米粒子功能化石墨稀構(gòu)建的雙重信號(hào)放大型電化學(xué)免疫傳感器的研究該工作基于納米金功能化的多孔石墨稀復(fù)合材料(Au-PGO)作為抗體的固定化基質(zhì),利用Au@Pd雙金屬納米粒子修飾的石墨烯(Au@Pd-Gra)標(biāo)記生物分子制備信號(hào)探針,構(gòu)建了一種用于檢測(cè)胰腺癌腫瘤標(biāo)志物(CA19-9)的夾心型電化學(xué)免疫傳感器。一方面,Au-PGO不僅可以固載大量的抗體生物分子,還能有效增加電極的活性面積,提高電極和氧化還原探針之間的電子傳輸能力；另一方面,Au@Pd-Gra不僅本身具有過(guò)氧化物模擬酶性質(zhì),其較大的比表面積和優(yōu)良的生物相容性等特點(diǎn),還可以用來(lái)固載更多的生物分子并保持生物分子的活性。同時(shí),在Au@Pd-Gra和HRP的協(xié)同催化作用下,電化學(xué)信號(hào)實(shí)現(xiàn)了雙重信號(hào)放大,免疫傳感器的靈敏度得到了極大的提高。在優(yōu)化的實(shí)驗(yàn)條件下,免疫傳感器對(duì)目標(biāo)分析物的檢測(cè)范圍為0.015-150 U·mL-1,最低檢測(cè)限為0.006 U·mL-1。該方法操作簡(jiǎn)便、靈敏度高,在定量檢測(cè)的臨床分析中具有很大的應(yīng)用潛力。
[Abstract]:In the analysis of medical immunoassay, the development of rapid, sensitive and accurate detection methods is the focus of research workers and hot issues. This is of great significance for the diagnosis of disease in clinical analysis. Electrochemical immunosensor is a combination of electrochemical technology, sensor technology and immunoassay technology three. In recent decades, the development of new functionalized nanomaterials, the exploration of biomolecular immobilization technology and the construction of signal enhancement techniques have attracted wide attention in recent decades. Mainly from the preparation of functional nanomaterials, the construction of biomimetic interfaces and the innovation of signal amplification technology, a series of highly sensitive electrochemical immunosensor are prepared. The main contents are as follows: 1. electrochemistry immunity of multi walled carbon nanometers based on gold nanoparticles A sandwich type electrochemical immunosensor based on nano gold / (3- mercapto propyl) trimethoxy silane (Au/MPTS) modified sensor interface and nano gold functionalized multi walled carbon nanotube composite biomolecules used for signal probe was constructed for the study of tumor marker alpha fetoprotein (AFP), a tumor marker of liver cancer. First, nano gold was prepared by PDDA covalent crosslinking on the surface of multi walled carbon nanotubes to obtain nano gold functionalized carbon nanomaterials. Then, the antibody biologically and horseradish peroxidase (HRP) labeled by the electroactive active substance (Thi) was immobilized on the surface, and nanoscale probes were prepared. The multi walled carbon nanotubes hold a large number of biomolecules and electroactive substances, which can effectively improve the sensitivity of the immunosensor. The nanoscale probe can not only identify the target analyte, but also further amplify the electrochemical reaction signal under the catalysis of HRP to the substrate H2O2. In addition, the interface of the Au/MPTS modified sensor is not It can only be used to immobilizing biomolecules, maintaining their biological activity, and further promoting electron transfer. The quantitative detection of AFP by sandwich immunoreaction model is realized. The linear range is 0.01-50.0 ng. ML-1, the detection limit is 3 pg. ML-1, the sensor has good selectivity, stability and reproducibility.2. based on double enzyme catalytic precipitation. In this experiment, a novel electrochemical immunosensor was constructed with carbon nanometers and double enzyme catalyzed product precipitation in this experiment. The high sensitivity detection of AFP was realized. First, the preparation of gold graphite thin composite nanomaterial (Au-Gra) was used in modified glassy. Carbon electrode can not only promote the electron transfer of the redox probe and the surface of the electrode, but also increase the effective area of the electrode to increase the solid load of the AFP antibody. Secondly, the carbon nano angle after the functionalized treatment is used to mark the antibody and the immobilized glucose oxidase (GOD) and HRP to get the nano probe, and the specific surface area of the carbon nano angle is increased. Using the sandwich immunoreaction model and using 4- naphthol as the precipitation substrate, the probe can not only identify the target analyte, but also enlarge the sensitivity of the immune sensor by double enzyme catalyzed precipitation, so as to realize the high sensitivity detection of the tumor marker. Characterization with electrochemical impedance and cyclic voltammetry. The linear response range of the immune sensor to the target analyte AFP is 0.001-60 ng. ML-1, and the detection limit is 0.33pg-mL-13. based on the dual signal amplified electrochemical immunosensor based on the nuclear shell structure Au@Pd bimetallic nanoparticles. Porous graphite thin composite (Au-PGO) is used as a immobilized matrix for antibody, and Au@Pd bimetal nanoparticles modified graphene (Au@Pd-Gra) biomolecules are used to prepare signal probes. A sandwich electrochemical immunosensor for detecting tumor markers (CA19-9) of pancreatic cancer (CA19-9) is constructed. On one hand, Au-PG O can not only immobilizing a large number of antibody biologic molecules, but also effectively increasing the active area of the electrode and improving the electronic transmission ability between the electrode and the redox probe. On the other hand, Au@Pd-Gra not only has the properties of the peroxidase mimic enzyme, but also the larger surface area and the excellent biocompatibility and so on. At the same time, under the synergistic catalysis of Au@Pd-Gra and HRP, the electrochemical signal has double signal amplification and the sensitivity of the immune sensor has been greatly improved. Under the optimized experimental conditions, the detection range of the immune sensor to the target analysis is 0.015-150 U. ML-1. The minimum detection limit is 0.006 U mL-1.. This method is simple and sensitive, and has great potential in clinical analysis of quantitative detection.

【學(xué)位授予單位】：西南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：TP212;TB383.1

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 蔡新霞,李華清,饒能高,王利,崔大付;電化學(xué)生物傳感器[J];微納電子技術(shù);2003年Z1期

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本文編號(hào)：1789934