本文關(guān)鍵詞： 電化學(xué)發(fā)光 免疫傳感器 魯米諾 納米復(fù)合材料 腫瘤標(biāo)志物　出處：《上海師范大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：電化學(xué)發(fā)光技術(shù)是發(fā)光物質(zhì)在電極表面通過電子得失產(chǎn)生的化學(xué)發(fā)光的現(xiàn)象,該技術(shù)能夠?qū)㈦娔苤苯愚D(zhuǎn)化為光能。近年來,電化學(xué)發(fā)光技術(shù)受到越來越多研究人員的關(guān)注,在藥物分析、免疫分析、DNA分析、臨床分析等科學(xué)領(lǐng)域得到廣泛運(yùn)用。新型功能性納米復(fù)合材料由于其特殊的納米結(jié)構(gòu)、較強(qiáng)的吸附能力、大的比表面積等優(yōu)勢(shì)為電化學(xué)傳感器的研究和應(yīng)用提供新途徑。將其應(yīng)用到電化學(xué)發(fā)光免疫傳感器的構(gòu)建中,可大大提高生物分子(如酶、DNA等)的固載量、標(biāo)記生物分子、催化反應(yīng)、加快電子傳遞,增大電流信號(hào)等。本論文以將多壁碳納米管-鉑-魯米諾(MWCNTs-Pt-luminol)作為免疫平臺(tái),構(gòu)建超靈敏的電化學(xué)發(fā)光免疫傳感器檢測(cè)糖抗原19-9(CA19-9);以MoS2-PEI-Au和Au@BSA兩種功能性納米材料分別作為傳感平臺(tái)和標(biāo)記魯米諾(luminol),構(gòu)建靈敏的電化學(xué)發(fā)光免疫傳感器檢測(cè)甲胎蛋白(AFP);以luminol標(biāo)記的Pt@BSA納米粒子作為包被抗原的固載基質(zhì),利用介孔碳球-二茂鐵(MCS-Fc)納米材料并對(duì)傳感器界面進(jìn)行修飾,用來檢測(cè)糖抗原15-3(CA15-3)。本論文主要包括四個(gè)方面內(nèi)容:1.闡述了論文的背景和意義,主要介紹了電化學(xué)發(fā)光的機(jī)理和功能性納米材料在電化學(xué)發(fā)光傳感器中的應(yīng)用,電化學(xué)免疫傳感器在實(shí)際生活中的應(yīng)用等。2.合成MWCNTs-Pt-luminol納米復(fù)合材料,借助紫外可見光譜和透射電鏡進(jìn)行表征。將上述功能性材料進(jìn)行組裝構(gòu)建了超靈敏的電化學(xué)免疫傳感器,并用于檢測(cè)CA19-9。本方法最低檢出限為0.000046 U/mL,具有良好的重現(xiàn)性及穩(wěn)定性,在實(shí)際血清樣品測(cè)定中的結(jié)果令人滿意。3.膠體金功能化的MoS2納米片通過靜電作用合成MoS2-PEI-Au納米復(fù)合材料固定在電極表面作為傳感平臺(tái),Au@BSA提供了更大的比表面積用來標(biāo)記luminol和固定二抗(Ab2),利用抗原與抗體之間的特異性結(jié)合構(gòu)建了靈敏檢測(cè)甲胎蛋白的夾心型電化學(xué)發(fā)光免疫傳感器。在最佳的實(shí)驗(yàn)條件下,該傳感器對(duì)AFP檢測(cè)的線性范圍從0.0001-200.0ng/m L,檢測(cè)限為0.00001ng/m L。此外,該傳感器對(duì)實(shí)際樣品的檢測(cè)顯示出優(yōu)良的性能,為其他臨床生物標(biāo)志物的檢測(cè)提供了巨大的潛力。4.利用具有大的比表面積、良好的導(dǎo)電性以及催化活性的介孔碳球-二茂鐵(MCS-Fc)作為基底材料,可以催化H2O2分解,實(shí)現(xiàn)電化學(xué)發(fā)光信號(hào)的增強(qiáng)。luminol-Pt@BSA-Ab2作為信號(hào)探針,當(dāng)孵育CA15-3之后,從而成功構(gòu)建了一個(gè)信號(hào)放大的夾心型電化學(xué)發(fā)光免疫傳感器,并實(shí)現(xiàn)了對(duì)CA15-3的超靈敏檢測(cè)。該免疫傳感器具有較寬的線性范圍和較低的檢測(cè)線。在最佳的實(shí)驗(yàn)條件下,該傳感器對(duì)CA15-3檢測(cè)的線性范圍從0.0005-500.0 U/mL,檢測(cè)限為0.000025 U/m L。此外,該免疫傳感器的構(gòu)建思路為臨床上其他生物分子和蛋白質(zhì)的檢測(cè)提供了一定的參考價(jià)值。
[Abstract]:Electrochemiluminescence (ECL) is a phenomenon of chemiluminescence produced by electroluminescent materials passing through electrons on the electrode surface. This technology can convert electric energy directly into light energy. In recent years, more and more researchers have paid attention to ECL technology. New functional nanocomposites are widely used in the fields of drug analysis, immunoassay, DNA analysis, clinical analysis and so on. The advantages of large specific surface area provide a new way for the research and application of electrochemical sensors. Applying them to the construction of electrochemiluminescence immunosensors can greatly increase the immobilization capacity of biomolecules (such as enzyme DNA) and label biomolecules. In this paper, we take MWCNTs-Pt-luminolol as the immune platform, which is based on multiwalled carbon nanotubes (MWCNTs-Pt-luminolol). A super sensitive electrochemiluminescence immunosensor was constructed to detect sugar antigen 19-9 and CA19-9, and two functional nanomaterials, MoS2-PEI-Au and Au@BSA, were used as sensing platform and labeled luminol, respectively, to construct a sensitive electrochemiluminescence immunosensor for the detection of nail fetal eggs. Luminol labeled Pt@BSA nanoparticles were used as the immobilized matrix for encapsulating the antigen. The mesoporous carbon-ferrocene (MCS-Fc) nanomaterials were used to detect carbohydrate antigens 15-3CCA15-3 by modifying the sensor interface. This paper mainly includes four aspects: 1.The background and significance of the paper are described. The mechanism of electrochemiluminescence and the application of functional nano-materials in electroluminescent sensors, the application of electrochemical immunosensors in practical life, etc. 2. Synthesis of MWCNTs-Pt-luminol nanocomposites are introduced in this paper. Ultraviolet-visible spectroscopy and transmission electron microscope were used to characterize the electrochemical immunosensor. The ultrasensitive electrochemical immunosensor was assembled and used for the detection of CA19-9.The detection limit of the method was 0.000046 UmL, which showed good reproducibility and stability. The results obtained in the actual serum samples are satisfactory .3. Colloidal gold-functionalized MoS2 nanoparticles were synthesized by electrostatic interaction to fix MoS2-PEI-Au nanocomposites on the electrode surface as a sensing platform, providing a larger specific surface area to be used as a sensing platform. A sandwich electrochemiluminescence immunosensor for sensitive detection of alpha-fetoprotein was constructed by labeling luminol and immobilized second antibody Ab2H. Under the optimum experimental conditions, a sandwich electrochemiluminescence immunosensor was constructed for the detection of alpha-fetoprotein by specific binding between antigens and antibodies. The linear range of the sensor for AFP detection is 0.0001-200.0ngr / mL, and the detection limit is 0.00001ngr / mL. in addition, the sensor shows excellent performance for the detection of real samples. It provides a great potential for the detection of other clinical biomarkers. Using MCS-Fc with large specific surface area, good electrical conductivity and catalytic activity as substrate material, it can catalyze the decomposition of H _ 2O _ 2. The enhancement of electrochemiluminescence signal. Luminol-Ptr BSA-Ab2 was used as a signal probe. After incubating CA15-3, a sandwich electroluminescent immunosensor with amplified signal was successfully constructed. The sensitivity detection of CA15-3 is realized. The immunosensor has a wide linear range and a lower detection line. Under the optimum experimental conditions, the linear range of detection for CA15-3 is 0.0005-500.0 UmLand the detection limit is 0.000025 U / mL. The construction of the immunosensor provides a certain reference value for the detection of other biomolecules and proteins in clinic.
【學(xué)位授予單位】：上海師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：O657.1

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相關(guān)期刊論文 前2條

1 Zanzan Zhu;;An Overview of Carbon Nanotubes and Graphene for Biosensing Applications[J];Nano-Micro Letters;2017年03期

2 Lili Yu;Hui Wu;Beina Wu;Ziyi Wang;Hongmei Cao;Congying Fu;Nengqin Jia;;Magnetic Fe_3O_4-Reduced Graphene Oxide Nanocomposites-Based Electrochemical Biosensing[J];Nano-Micro Letters;2014年03期

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本文編號(hào)：1497623