本文關(guān)鍵詞：基于APTES和蛋白A聯(lián)合固定化技術(shù)的乙肝病毒免疫生物傳感器敏感膜的制作研究　出處：《浙江大學(xué)》2012年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： APTES硅烷化 定向固定 原子力顯微鏡(AFM) 免疫傳感器

【摘要】：穩(wěn)定、高效的生物微納免疫傳感器在食品領(lǐng)域、環(huán)境監(jiān)測領(lǐng)域及醫(yī)療診斷領(lǐng)域的廣泛應(yīng)用。生物傳感器敏感膜固定化技術(shù)是生物傳感器的關(guān)鍵核心技術(shù)。傳統(tǒng)敏感膜固定方法(吸附法、共價(jià)結(jié)合法、交聯(lián)法以及包埋法等)漸趨成熟但普遍存在固定生物分子失活,效價(jià)不高的問題。生物大分子定向固定可以提高生物材料固定的有效性,但是卻存在牢固性和可重用性差等問題。 針對上述問題,本文以乙肝免疫生物傳感器敏感膜固定化技術(shù)研究為切入點(diǎn),首先,提出了將傳統(tǒng)化學(xué)共價(jià)結(jié)合和生物大分子定向固定化方法結(jié)合的生物傳感器敏感膜固定化技術(shù),它既利用了化學(xué)共價(jià)固定法的固定牢固性和特異性,又利用了蛋白八定向固定的抗空間位阻性和有效性高的特性。文中的化學(xué)共價(jià)修飾采用了能夠有效控制修飾的APTES硅烷化固定方法,并實(shí)現(xiàn)了對于基底膜性質(zhì)的有效控制。其次,為進(jìn)一步提高傳感器靈敏度等方面的性能,我們對固定化界面進(jìn)行了納米材料的修飾,通過控制材料表面納米顆粒的大小、性質(zhì)及反應(yīng)條件,有效控制了納米材料的納米層面上的性質(zhì),進(jìn)而有效利用了修飾材料的納觀效應(yīng)。為驗(yàn)證上述固定化方法的有效性,我們通過接觸角(WC八)傅里葉變換紅外線光譜(FTIR)檢測等實(shí)驗(yàn)方法定性驗(yàn)證了敏感膜的修飾結(jié)果,并用原子力顯微鏡對修飾過程進(jìn)行了形貌學(xué)表征。 首先,通過對比分析研究在經(jīng)過或未經(jīng)過APTES硅烷化修飾的硅片上固定蛋白八的效果,證明了APTES硅烷化修飾的優(yōu)越性;其次,分析比較了通過APTES硅烷化納米顆粒修飾后僑聯(lián)蛋白A或不僑聯(lián)蛋白A的兩種方法,進(jìn)一步證明蛋白八對于本固定方法中抗體定向固定,降低空間位阻,提高傳感靈敏度的重要性。最后,通過雙抗體夾心ELISA方法定性驗(yàn)證了上述聯(lián)合固定化方法在微量固定抗體情況下的高活性。上述結(jié)果表明了通過將聯(lián)合固定化技術(shù)與界面納觀效應(yīng)有機(jī)結(jié)合可實(shí)現(xiàn)生物免疫傳感器敏感膜的良好固定。
[Abstract]:Stable and efficient biosensors for biosensors in the field of food. Biosensor sensitive membrane immobilization technology is the key technology of biosensor. Traditional sensitive membrane fixation methods (adsorption method, covalent binding method) are widely used in the field of environmental monitoring and medical diagnosis. Crosslinking method and embedding method, etc.) but the problem of immobilization of biomolecules is common, such as inactivation and low titer of biomolecules. Directional fixation of biomolecules can improve the efficiency of biomaterials fixation. However, there are problems such as poor fastness and reusability. In order to solve the above problems, this paper focuses on the study of immobilization of sensitive membrane of hepatitis B immunosensor. The biosensor sensitive membrane immobilization technology which combines the traditional chemical covalent binding method and the biological macromolecule directional immobilization method is proposed. It not only makes use of the fixation fastness and specificity of the chemical covalent fixation method. The chemical covalent modification is based on the APTES silanization method which can effectively control the modification. And realized the effective control of the properties of the substrate membrane. Secondly, in order to further improve the sensor sensitivity and other aspects of the performance, we modified the immobilized interface nanomaterials. By controlling the size, properties and reaction conditions of nanocrystalline particles on the surface of the materials, the properties of the nanomaterials on the nanometer level are effectively controlled. Furthermore, the nano effect of the modified materials is effectively utilized to verify the effectiveness of the above immobilization methods. The modification results of the sensitive films were qualitatively verified by means of FTIR (contact angle WC8) Fourier transform infrared spectroscopy (FTIR). The modification process was characterized by atomic force microscope (AFM). Firstly, by comparing and analyzing the effect of immobilization of protein 8 on the silicon wafer modified with or without APTES silanization, the superiority of APTES silanization modification was proved. Secondly, two methods modified by APTES silanized nanoparticles were analyzed and compared. It was further proved that protein 8 was immobilized against antibodies in this method. Reduce the space steric resistance, improve the importance of sensing sensitivity. Finally. The double antibody sandwich ELISA method was used to qualitatively verify the high activity of the combined immobilization method in the case of microimmobilized antibodies. The above results show that the joint immobilization technique and the interface nano effect are organic junctions. It can achieve good fixation of biosensor sensitive membrane.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R373;TP212.3

【共引文獻(xiàn)】

相關(guān)期刊論文 前1條

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相關(guān)碩士學(xué)位論文 前3條

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3 邱思遠(yuǎn);納米金阻止VEGF165與VEGFR2結(jié)合的原子力顯微鏡研究[D];暨南大學(xué);2010年

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本文編號(hào)：1383542