發(fā)布時(shí)間：2018-08-13 12:12

【摘要】：人絨毛膜促性腺激素(h CG)是一種由胎盤絨毛膜囊泡的滋養(yǎng)層細(xì)胞分泌的糖蛋白激素,屬于肽類激素,由α亞基、β亞基和244中氨基酸組成,分子量為36.7 k Da。hCG存在于人體尿液和血液中。女性在懷孕期間,hCG在體內(nèi)含量變化與妊娠期的一些疾病有關(guān),比如卵巢癌、絨毛上皮癌等,所以構(gòu)建一種能夠快速方便的檢測hCG的方法是非常有意義而且很急迫的。電化學(xué)免疫傳感器具有很多優(yōu)點(diǎn),比如高靈敏度、成本低、易操作、處理步驟簡單以及檢測迅速等,是一種理想的方法。電極的修飾是影響構(gòu)建的電化學(xué)免疫傳感器的電化學(xué)響應(yīng)的關(guān)鍵步驟,同時(shí)影響生物分子的固定。由于納米材料具有比表面積大、生物相容性好、電子傳遞速率快、較高的催化活性以及較強(qiáng)的吸附能力等諸多優(yōu)點(diǎn),是修飾電極的理想材料。本論文利用具有不同形貌和不同結(jié)構(gòu)的納米材料修飾電極,包括多壁碳納米管、納米多孔Pd和納米金,構(gòu)建電化學(xué)免疫傳感器,提高電化學(xué)免疫傳感器的靈敏度,改善電化學(xué)免疫傳感器的重現(xiàn)性、穩(wěn)定性以及特異性,以滿足臨床上對一些生物分子的檢測。1.基于多壁碳納米管-室溫離子液體的納米復(fù)合物和納米多孔Pd構(gòu)建電化學(xué)免疫傳感器對h CG進(jìn)行檢測多壁碳納米管(MWCNTs)和室溫離子液體(RTIL)-1-丁基-3-甲基咪唑六氟磷酸鹽(BMIMPF6)組成的復(fù)合物膜作為電極的第一層修飾材料,這種復(fù)合物材料結(jié)合了MWCNTs和RTIL的優(yōu)點(diǎn)。同時(shí)討論其協(xié)同作用機(jī)理;其次,通過腐蝕得到的納米多孔Pd(NP-Pd)作為電極的第二層修飾材料,NP-Pd的結(jié)構(gòu)通過EDS、XRD、SEM、TEM和BET進(jìn)行表征。由于NP-Pd具有較高的比表面積和優(yōu)異的導(dǎo)電性,hCG的固定量和電子轉(zhuǎn)移速率都得到很大的提高。在優(yōu)化的實(shí)驗(yàn)條件下,免疫傳感器的線性范圍為0.05-50 ng m L-1,檢測限為3.2 pg mL-1。構(gòu)建的免疫傳感器具有較高的靈敏度、良好的重現(xiàn)性和穩(wěn)定性。此外,將構(gòu)建的免疫傳感器用于實(shí)際樣品進(jìn)行檢測,結(jié)果令人滿意。2.利用納米多孔Pd做標(biāo)記構(gòu)建 三明治‖型電化學(xué)免疫傳感器對hCG進(jìn)行檢測通過腐蝕的方法得到NP-Pd,由于NP-Pd具有優(yōu)異的導(dǎo)電性、良好的生物相容性以及強(qiáng)的電化學(xué)催化活性,利用NP-Pd標(biāo)記辣根過氧化酶標(biāo)記的抗體(HRP-Ab2)(NP-Pd-HRP-Ab2)構(gòu)建 三明治‖型電化學(xué)免疫傳感器。納米金(GNPs)、普魯士藍(lán)(PB)和GNPs作為電極修飾基質(zhì),不僅用于固定抗體(Ab1),而且對信號放大具有很大的作用。在優(yōu)化的實(shí)驗(yàn)條件下,免疫傳感器的線性范圍為0.05-200 ng mL-1,檢測限為9.2 pg mL-1。構(gòu)建的免疫傳感器具有較高的靈敏度、良好的重現(xiàn)性和穩(wěn)定性。此外,構(gòu)建的免疫傳感器用于實(shí)際樣品進(jìn)行檢測,結(jié)果令人滿意。
[Abstract]:Human chorionic gonadotropin (h CG) is a glycoprotein hormone secreted by trophoblastic cells of placental chorionic vesicles. The molecular weight of 36.7 k Da.hCG is present in human urine and blood. The changes of hCG in women during pregnancy are related to some diseases during pregnancy, such as ovarian cancer and choriocarcinoma, so it is very meaningful and urgent to construct a rapid and convenient method to detect hCG. Electrochemical immunosensor has many advantages, such as high sensitivity, low cost, easy operation, simple processing and rapid detection. Electrode modification is a key step to affect the electrochemical response of the constructed electrochemical immunosensor, as well as the immobilization of biomolecules. Nanomaterials are ideal materials for modified electrodes because of their large specific surface area, good biocompatibility, fast electron transfer rate, high catalytic activity and strong adsorption ability. In this paper, electrochemical immunosensors were constructed by using nano-materials with different morphologies and structures, including multi-walled carbon nanotubes, nano-porous PD and nano-gold, to improve the sensitivity of electrochemical immunosensors. To improve the reproducibility, stability and specificity of electrochemical immunosensors for clinical detection of some biomolecules. Electrochemical immunosensor based on nanocomposites of multi-walled carbon nanotubes and room temperature ionic liquids and nano-porous PD for the detection of hCG (MWCNTs) and room temperature ionic liquids (RTIL) -1- Ding Ji -3-methylimidazolium hexafluorophosphoric acid The composite membrane composed of salt (BMIMPF6) was used as the first layer modification material of the electrode. This composite material combines the advantages of MWCNTs and RTIL. At the same time, the synergistic mechanism of NP-Pd was discussed. Secondly, the structure of nano-porous PD (NP-Pd) as the second layer modified material was characterized by BET and BET. The fixed amount and electron transfer rate of NP-Pd have been greatly improved because of its high specific surface area and excellent conductivity. Under the optimized experimental conditions, the linear range of the immunosensor was 0.05-50 ng mL ~ (-1) and the detection limit was 3.2 PG mL ~ (-1). The constructed immunosensor has high sensitivity, good reproducibility and stability. In addition, the constructed immunosensor was used to detect the actual samples with satisfactory results of. 2. 2. NP-Pd was prepared by using nano-porous PD as marker to construct a sandwich type electrochemical immunosensor for the detection of hCG. NP-Pd was obtained by corrosion method. Because of the excellent conductivity, good biocompatibility and strong electrochemical catalytic activity of NP-Pd, NP-Pd was obtained from the electrochemical immunosensor. A sandwich type electrochemical immunosensor was constructed using NP-Pd labeled horseradish peroxidase labeled antibody (HRP-Ab2) (NP-Pd-HRP-Ab2). Nanocrystalline gold (GNPs), Prussian blue (PB) and GNPs were used as electrode modified matrix not only for immobilization of antibody (Ab1) but also for signal amplification. Under the optimized experimental conditions, the linear range of the immunosensor was 0.05-200 ng mL -1 and the detection limit was 9.2 PG mL -1. The constructed immunosensor has high sensitivity, good reproducibility and stability. In addition, the constructed immunosensor was used to detect the actual samples with satisfactory results.
【學(xué)位授予單位】：濟(jì)南大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：TB383.1;TP212

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