本文關(guān)鍵詞： 電化學免疫傳感器 石墨烯量子點 包銅去鐵蛋白 磁性碳納米管 中空介孔二氧化硅　出處：《山東農(nóng)業(yè)大學》2015年碩士論文　論文類型：學位論文

【摘要】：禽白血病是由不同的RNA逆轉(zhuǎn)錄病毒引起的可傳播性的家禽腫瘤性疾病。近幾年來,由禽白血病引起的家禽死亡及淘汰率逐年上升,造成的經(jīng)濟損失逐年增加。除此之外,由禽白血病感染之后造成的免疫抑制和對疫苗的應答能力下降等間接損失更是無法估量。本文研究了石墨烯量子點、包銅去鐵蛋白、磁性碳納米管和中空介孔二氧化硅等納米材料及其復合材料的電化學性能,并結(jié)合分子識別、直接氧化還原、納米材料的類酶催化等方法,構(gòu)建了幾種不同類型的電化學免疫傳感器,實現(xiàn)了對禽白血病病毒的超靈敏檢測。主要分為以下三部分:(1)本研究用石墨烯量子點和包銅去鐵蛋白作為信號放大物質(zhì),制作了一種新穎的檢測禽白血病病毒的電化學免疫傳感器。分別以石墨烯量子點修飾電極連接一抗,復合四氧化三鐵磁珠連接二抗,實現(xiàn)三明治式的電極組裝。然后用包銅去鐵蛋白結(jié)合Fe3O4@GQDs納米粒子,將電信號活性物質(zhì)-銅納米粒子引入了傳感器體系中。當改變環(huán)境的pH值,電信號物質(zhì)被釋放到溶液中,可以通過微分脈沖伏安法對釋放出來的電信號物質(zhì)進行檢測。由于石墨烯量子點有巨大的比表面積和大量的含氧官能團,可以結(jié)合更多的抗體,從而放大體系的電化學信號。去鐵蛋白提供了容納信號物質(zhì)的巨大空腔,使得檢測信號進一步被放大。實驗中,通過對比不同標記的信號探針,證實了傳感器的信號放大作用。禽白血病病毒濃度在102.08到104.50 TCID50/mL范圍內(nèi)呈很好的線性關(guān)系,在信噪比為3時,檢測限為115 TCID50/mL。實驗結(jié)果顯示該傳感器具有高的靈敏度,好的重復性和穩(wěn)定性。(2)本研究首次以中空介孔二氧化硅和去鐵蛋白聯(lián)合構(gòu)建了一種受pH控制的新型免疫傳感器,實現(xiàn)了對禽白血病病毒的高靈敏檢測。首先,制備了環(huán)糊精功能化的石墨烯,中空介孔二氧化硅、包銅去鐵蛋白和二抗的復合物(HMS-apoferritin-Cu/Ab2-BSA),利用TEM對CD-GS、HMS和HMS-apoferritin-Cu/Ab2-BSA探針進行了表征。然后,以CD-GS作為基底、HMS-apoferritin-Cu/Ab2-BSA作為探針構(gòu)建電化學免疫傳感器。利用CV對修飾電極進行了表征,研究了CD-GS的電化學性能,探討了富集時間和富集電位對響應信號的影響。最后利用DPV實現(xiàn)了對ALVs-J的高靈敏檢測,并探討了免疫傳感器的選擇性、重現(xiàn)性和穩(wěn)定性。(3)基于一種多重還原銀的信號放大方法,構(gòu)建了一種三明治式的電化學免疫傳感器,實現(xiàn)了對禽白血病病毒的檢測。首先,用CD-GS修飾的裸電極連接一抗,然后用牛血清白蛋白進行活性位點的封閉,以防止非特異性的吸附;經(jīng)過三明治式的電極組裝,CIMC-Au-HAs/Ab2-BSA探針被引入到免疫傳感器中,再把組裝好的電極放入含銀離子的溶液中進行銀的還原,還原出來的銀通過微分脈沖伏安法進行檢測。為了說明磁性碳納米管優(yōu)越的性能,納米金的催化還原能力和腐殖酸的優(yōu)良吸附能力,我們對不同的納米探針進行了比較。禽白血病病毒濃度在102.05到104.50 TCID50/mL范圍內(nèi)呈很好的線性關(guān)系,在信噪比為3時,檢測限為110 TCID50/mL。實驗結(jié)果顯示該傳感器具有高的靈敏度,潛在的臨床應用能力。
[Abstract]:Avian leukosis is caused by different RNA retrovirus can spread the avian tumor diseases. In recent years, caused by avian leukosis poultry deaths and elimination rate increased year by year, the economic losses caused by increased year by year. In addition, caused by avian leukosis virus infection after immune suppression and vaccine response ability the decline of indirect losses is incalculable. This paper studied graphene quantum dots, copper coated iron protein, the electrochemical properties of carbon nanotubes and hollow mesoporous silica nanoparticles and their composite materials, and combined with molecular recognition, direct redox, nano materials enzymatic method, constructed electrochemical immunoassay several different types of sensors, to achieve the ultra sensitive detection of avian leukosis virus. Mainly divided into the following three parts: (1) this study used as graphene quantum dots and copper coated iron protein The signal amplification material, electrochemical immunosensor fabricated a novel avian leukosis virus detection. With graphene quantum dot modified electrode is connected with a resistance, connecting the two anti magnetic composite ferroferric oxide electrode, realize the sandwich assembly. Then use the copper to iron binding protein of Fe3O4@GQDs nanoparticles, the electrical signal of active substances - copper nanoparticles the introduction of the sensor system. When changing the pH value of the environment, physical signals are released into the solution, by differential pulse voltammetry to detect signal substances released. Because graphene quantum dots have huge surface area and a large number of oxygen-containing functional groups, can be combined with more antibodies, thereby amplifying the electrochemical signal system. To provide a huge iron protein cavity containing signal substances, so the detection signal is further amplified. In the experiment, by comparing with standard Remember that a signal probe, the sensor signal amplification. Avian leukosis virus concentration showed a good linear relationship in the range of TCID50/mL 102.08 to 104.50, when the signal-to-noise ratio is 3, the detection limit was 115 TCID50/mL.. The experimental results show that the sensor has high sensitivity, good repeatability and stability. (2) this is the first study to hollow mesoporous silica and iron protein and constructed a new immune sensor controlled by pH, the realization of avian leukosis virus sensitive detection. Firstly, cyclodextrin functionalized graphene prepared hollow mesoporous silica, copper clad iron protein and complex two - the use of CD-GS (HMS-apoferritin-Cu/Ab2-BSA), TEM, HMS and HMS-apoferritin-Cu/Ab2-BSA probe were investigated. Then, using CD-GS as substrate, HMS-apoferritin-Cu/Ab2-BSA as a probe to construct electrochemical immunosensor. The modified electrode was characterized by CV, the electrochemical properties of CD-GS were investigated and discussed the enrichment time and accumulation potential of the response signal. Finally realizes the high sensitive detection of ALVs-J by DPV, and discusses the selective immune sensor, reproducibility and stability. (3) a multiple signal reduction of silver amplification method based on the construction of a sandwich type electrochemical immunosensor, realized the detection of avian leukosis virus. First of all, the bare electrode modified with CD-GS connection of a resistance, then the active site of bovine serum albumin closed, to prevent nonspecific adsorption; after the electrode sandwich assembly, CIMC-Au-HAs/Ab2-BSA probe is introduced to the immune sensor, then restore the assembled electrode into the solution containing silver ion in silver, the reduction of silver by differential pulse voltammetry for detection. The magnetic properties of carbon nanotubes superior catalytic reduction of gold nanoparticles, excellent adsorption ability and humic acid, we compare the different nano probe. Avian leukosis virus concentration showed a good linear relationship in the range of TCID50/mL 102.05 to 104.50, when the signal-to-noise ratio is 3, the detection limit is 110 TCID50/mL. the experimental results show that the sensor has high sensitivity, clinical application potential.

【學位授予單位】：山東農(nóng)業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S852.65;TB383.1

【共引文獻】

相關(guān)博士學位論文 前2條

1 周亞平;基于壓電電化學和色譜電化學的生化分析[D];湖南師范大學;2014年

2 杭柏林;J亞群禽白血病病毒JS09GY3株感染特性及感染雞法氏囊轉(zhuǎn)錄組學分析[D];揚州大學;2014年

相關(guān)碩士學位論文 前2條

1 辛敬凱;B亞群禽白血病病毒單克隆抗體的制備[D];山東農(nóng)業(yè)大學;2014年

2 單文倩;基于壓電生物傳感的腫瘤細胞檢測及納米顆粒細胞毒性研究[D];湖南大學;2014年

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本文編號：1546300