本文選題：半乳糖氧化酶 + 石墨烯　； 參考：《湘潭大學(xué)》2017年碩士論文

【摘要】：由于第三代酶生物傳感器在醫(yī)療和環(huán)境方面的良好的應(yīng)用前景,與第三代生物傳感器的相關(guān)課題引起了科研人員的巨大興趣。21世紀(jì)開始的階段,材料科學(xué)方面取得的進步,給第三代生物傳感器帶來了新的機遇。納米材料以及導(dǎo)電聚合物,改進了第三代生物傳感器的一些相關(guān)性質(zhì)。其中,石墨烯的導(dǎo)電性比以往的納米材料有所提高,比表面積較大,適合生物分子的固定。有助于生物分子活性的保留。本文制作了三種性能良好的的第三代生物傳感器,利用功能化石墨烯等材料固定血藍蛋白,半乳糖氧化酶,對所制備的生物傳感器進行了研究。主要內(nèi)容為:使聚乙二醇乙二酸脂(Poly(ethylene succinate),PES)與氧化石墨烯(GO)結(jié)合。對氧化石墨烯(GO)在-0.60 V的電壓下進行電還原處理,獲得還原氧化石墨烯與PES復(fù)合材料,首次利用這種RGO/PES復(fù)合膜固載了血藍蛋白(hemocyanin,HC),形成了生物復(fù)合膜RGO/PES/HC,我們用掃描電子顯微鏡(SEM)表征這種生物復(fù)合膜的表面形貌,電化學(xué)交流阻抗(EIS)以及循環(huán)伏安法(CV)等電化學(xué)測試方法也被用于測試本工作制備的RGO/PES/HC修飾電極的電化學(xué)相關(guān)性質(zhì)。獲得了在不同pH條件和掃速的情況下的循環(huán)伏安圖,通過分析循環(huán)伏安圖來研究納米膜復(fù)合膜的電子傳遞過程。同時,用CV和微分脈沖伏安法(DPV)來研究本工作中制備的RGO/PES/HC修飾電極對雙酚A(Bisphenol A,BPA)的催化情況。實驗結(jié)果表明,制作的生物傳感器對BPA的檢測的線性范圍為8.0×10-8M到5.5×10-5M,檢出限為3.4×10-8 M,并利用本工作制備的生物傳感器對塑料材料產(chǎn)品中的雙酚A進行了實樣檢測,回收率在可接受范圍之內(nèi),結(jié)果仍然是讓人滿意的。利用原位電化學(xué)聚合的方法,合成了導(dǎo)電聚合物聚苯胺(polyaniline,PANI)/RGO復(fù)合膜,同時在原位生成聚苯胺的過程中,利用靜電作用的原理,將半乳糖氧化酶(GAO)固定在RGO/PANI納米復(fù)合材料內(nèi),利用交流阻抗(EIS)和循環(huán)伏安法(CV)研究了不同修飾電極的電化學(xué)性質(zhì)。并且利用安培法,研究了復(fù)合膜對于半乳糖的催化行為,通過計算得到的檢測限為9.0×10-6 M。同時對實驗條件進行了優(yōu)化,我們發(fā)現(xiàn)在PH=7.0,電位為-0.50 V時,獲得最好的檢測效果。在測試所制備的生物傳感器的抗干擾能力方面,檢測了常見干擾物的干擾情況,情況令人滿意,將制備好的修飾電極在冰箱中保存15天后,仍然有92%的響應(yīng)電流。對穩(wěn)定性和重現(xiàn)性進行測試,發(fā)現(xiàn)本工作構(gòu)建的酶電化學(xué)生物傳感器的穩(wěn)定性和重現(xiàn)性良好。利用聚甲基丙烯酸甲酯(polymethyl methacrylate,PMMA)功能化氧化石墨烯(GO),將半乳糖氧化酶(Galactose oxidase,GAO)固定在GO/PMMA納米復(fù)合材料之內(nèi),用SEM研究了GO/PMMA/GAO生物復(fù)合膜的表面形貌,進一步利用CV研究了生物復(fù)合膜的電化學(xué)行為,并在不同的pH和掃速下進行了循環(huán)伏安測試,研究了電極表面的電化學(xué)行為,同時用時間電流法(i-t)研究了傳感器對半乳糖和二羥基丙酮(1,3-Dihydroxyacetone,DHA)的催化行為,其中對半乳糖催化的線性范圍為7.0×10-6 M到2.4×10-4 M,對二羥基丙酮的線性檢測范圍為8.0×10-6 M到4.5×10-4 M檢出限7.0×10-6 M,并且,通過重現(xiàn)性實驗說明GO/PMMA/GAO生物傳感器具有良好的重現(xiàn)性。
[Abstract]:Due to the good application prospect of the third generation enzyme biosensor in the medical and environmental aspects, the related topics of the third generation biosensors have aroused the great interest of the researchers in the.21 century. The progress in material science has brought new opportunities to the third generation biosensors. Some of the related properties of the third generation biosensors are improved. Among them, the conductivity of graphene is higher than that of the previous nanomaterials, and the specific surface area is larger. It is suitable for the immobilization of biomolecules. It is helpful for the retention of biological molecules. In this paper, three kinds of third generation biosensors with good performance have been made, using functional fossils and so on. The materials are immobilized on hemocyanin and galactose oxidase, and the biosensors are studied. The main contents are: combining Poly (ethylene succinate) and PES with graphene oxide (GO). The electrochemical reduction of graphene oxide (GO) under the electric pressure of -0.60 V is carried out, and the reduction of graphene oxide and PES is obtained. Materials, the hemocyanin (HC) was immobilized by the RGO/PES composite membrane for the first time, and the biological composite membrane RGO/PES/HC was formed. We used scanning electron microscopy (SEM) to characterize the surface morphology of the composite membrane. Electrochemical impedance (EIS) and cyclic voltammetry (CV) methods were also used to test the preparation of this work. The electrochemical properties of the RGO/PES/HC modified electrode have been obtained. The cyclic voltammograms are obtained at different pH conditions and scavenging conditions. By analyzing the cyclic voltammograms, the electron transfer process of the nanocomposite membranes is studied. At the same time, the RGO/PES/HC modified electrodes prepared in this work are studied by CV and differential pulse voltammetry (DPV) to the bisphenol A (Bis). The experimental results show that the linear range of the detection of BPA by biosensor is 8 * 10-8M to 5.5 x 10-5M, the detection limit is 3.4 * 10-8 M, and the biological sensor prepared by this work is used to detect the bisphenol A in the plastic material, and the recovery rate is within acceptable range, and the result is still in the phenol. It is satisfactory. The polyaniline (PANI) /RGO composite membrane was synthesized by in situ electrochemical polymerization. In the process of in situ formation of polyaniline, the galactose oxidase (GAO) was immobilized in RGO/PANI nanocomposites by electrostatic action, and the AC impedance (EIS) and cyclic voltammetry (CV) method were used. CV) the electrochemical properties of different modified electrodes were studied. The catalytic behavior of the composite membrane for galactose was studied by amperometric method. The detection limit of 9 x 10-6 M. was calculated and the experimental conditions were optimized. We found that the best detection effect was obtained at PH=7.0, when the potential was -0.50 V. With respect to the anti-interference ability of the sensor, the interference of common interferers is detected. The situation is satisfactory. The prepared modified electrode still has 92% response current for 15 days in the refrigerator. The stability and reproducibility are tested. It is found that the stability and reproducibility of the enzyme electrochemistry biosensor constructed in this work are good. With polymethyl methacrylate (PMMA) functionalized graphene oxide (GO), galactose oxidase (Galactose oxidase, GAO) was immobilized in GO/PMMA nanocomposite. The surface morphology of GO/PMMA/GAO biological composite membrane was studied by SEM, and the electrochemical behavior of the biological composite membrane was studied by CV. At the same pH and sweep speed, cyclic voltammetry was carried out to study the electrochemical behavior of the electrode surface. At the same time, the catalytic behavior of the sensor to galactose and two hydroxyacetone (1,3-Dihydroxyacetone, DHA) was studied by time current method (I-T). The linear range of the catalysis of galactose was 7 x 10-6 M to 2.4 x 10-4 M, to the line of two hydroxy acetone. The detection limit is 8 * 10-6 M to 4.5 * 10-4 M detection limit 7 * 10-6 M, and reproducibility test shows that GO/PMMA/GAO biosensor has good reproducibility.
【學(xué)位授予單位】：湘潭大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O657.1;TB33

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相關(guān)期刊論文 前2條

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本文編號：1978449