本文關(guān)鍵詞： 毛細(xì)管電泳 蛋白質(zhì)分離 毛細(xì)管涂層制備　出處：《青島大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

【摘要】：毛細(xì)管電泳是一種以高壓電場(chǎng)力作為動(dòng)力,在毛細(xì)管內(nèi)部進(jìn)行分離的新型的分離技術(shù)。毛細(xì)管電泳技術(shù)是一種高效率,高靈敏度,經(jīng)濟(jì)快速的生物大分子分離分析工具,對(duì)蛋白質(zhì)的分離分析更有很大的優(yōu)勢(shì),但是,分析過程中蛋白質(zhì)會(huì)吸附在石英毛細(xì)管內(nèi)壁使分離分析不夠準(zhǔn)確,為了調(diào)整電滲流和抑制涂層對(duì)蛋白質(zhì)的吸附,因此,需要對(duì)熔融石英管內(nèi)壁進(jìn)行涂層處理,越來越多的毛細(xì)管電泳涂層涌現(xiàn)出來,也是目前解決這個(gè)難題最主要和常用的方法,常用的毛細(xì)管涂層方法主要有化學(xué)鍵合涂層和物理吸附涂層,這兩種涂層方式都各自的用途與優(yōu)點(diǎn),但是兩種方法都有一定的局限性和缺點(diǎn),其中物理方法制備步驟簡潔,但是由于涂層不穩(wěn)定,分離效果很差,影響分離效果,而化學(xué)鍵合涂層穩(wěn)定性高,但過程復(fù)雜。制備出質(zhì)量與性能更加優(yōu)異的毛細(xì)管涂層實(shí)現(xiàn)蛋白質(zhì)更好的分離分析效果是該領(lǐng)域研究人員的又一需要突破的重點(diǎn)。本論文采用層層自組裝技術(shù)(LBL)將大眾使用的毒性大、污染性強(qiáng)的硅烷化試劑換做安全環(huán)保的水溶性感光高分子重氮樹脂,重氮樹脂因其自身的結(jié)構(gòu)與特征使其具有獨(dú)特的光熱交聯(lián)反應(yīng),與毛細(xì)管涂層新材料反映后可以形成穩(wěn)定穩(wěn)固的共價(jià)鍵,涂層的性能更加穩(wěn)定,對(duì)混合蛋白的分離具有很好的效果。本論文主要工作如下:1.使用感光高分子材料重氮樹脂(DR)作為偶聯(lián)劑,通過自組裝和共價(jià)連接樹枝狀高分子(CD-Dendrimer)制備毛細(xì)管涂層,重氮樹脂與樹枝狀高分子的首先通過離子鍵固定在毛細(xì)管內(nèi)壁,通過紫外光線照射后,離子鍵轉(zhuǎn)化為共價(jià)鍵,從而穩(wěn)定的固定在毛細(xì)管內(nèi)壁,用該種涂層來分離幾種混合蛋白質(zhì),呈現(xiàn)出良好的分離效果。2.基于層層自組裝(LBL)技術(shù),制備出重氮樹脂與氧化石墨烯(GO)的毛細(xì)管涂層,在毛細(xì)管內(nèi)壁兩種分子交替涂層多次,最后在紫外光的照射下,原本重氮樹脂與氧化石墨烯之間相互作用的較弱的離子鍵轉(zhuǎn)化為更加牢固的共價(jià)鍵,從而制備出多層的結(jié)構(gòu)可控的共價(jià)鍵合涂層,用來分離幾種蛋白質(zhì),分離效果理想,重現(xiàn)性與穩(wěn)定性也很好。3.選取親水性物質(zhì)β-環(huán)糊精(β-CD)做為分離蛋白材料,與重氮樹脂首先通過離子鍵在毛細(xì)管內(nèi)表面形成復(fù)合膜,起到涂覆毛細(xì)管的作用,然后將其放置于紫外線下照射,通過重氮樹脂獨(dú)特的光學(xué)反應(yīng),是離子鍵轉(zhuǎn)化為穩(wěn)定的共價(jià)鍵,毛細(xì)管涂層更加穩(wěn)定,不易脫落,抑制蛋白的吸附作用明顯,將制備好的共價(jià)鍵合涂層DR/β-CD毛細(xì)管涂層用于分離混合蛋白,效果很好。本論文所研究和制備的毛細(xì)管涂層是一種新的偶聯(lián)劑與新材料的結(jié)合,制備出的一種新型毛細(xì)管涂層,不僅利用了常用的兩種涂層技術(shù)的優(yōu)點(diǎn),也在一些局限性方面做了很多改善,操作非常簡單快速,涂層也很穩(wěn)定。用以分離混合蛋白來測(cè)試其性能,分離結(jié)果非常的好,其他測(cè)試結(jié)果也良好,說明我們所制備的毛細(xì)管涂層是穩(wěn)定有效的,為毛細(xì)管的進(jìn)一步發(fā)展提供了更多可能。
[Abstract]:Capillary electrophoresis is a kind of high voltage electric field force as power, new separation technology in the separation capillary inside. Capillary electrophoresis is a kind of high efficiency, high sensitivity, rapid separation and analysis tools of economic biological macromolecules, for protein separation and analysis of advantages, but more substantial, analysis in the process of protein adsorption on quartz the inner wall of the capillary separation analysis is not accurate enough, in order to adjust the electroosmotic flow and adsorption, inhibiting coating on protein therefore need coating of fused quartz tube, capillary tube electrophoresis coating increasingly emerged, is now to solve the problem of the most important and commonly used method, method of capillary coating are mainly used for chemical bonding physical adsorption coating and coating, coating the two way of their uses and advantages, but the two methods have certain limitations and The preparation method of physical weakness, the steps are simple, but because the coating is not stable, the separation effect is poor, affect the separation effect, and chemical bonding coating with high stability, but the process is complex. Preparation of capillary coating more excellent quality and achieve better performance of protein separation effect analysis is the research in this field and a the need to break the key. This paper adopted the self-assembly technology (LBL) will be the mass use of high toxicity and pollution of silanes and strong for safety and environmental protection of water soluble photosensitive polymer diazoresin diazoresin, due to the structure and characteristics of its own. It has unique thermal crosslinking reaction, and can reflect the new material after the capillary coating to form covalent bonds to stabilize, the performance of the coating is more stable, the separation of mixed protein has a very good effect. The main work of this thesis are as follows: 1. using photosensitive polymer Material diazoresin (DR) as a coupling agent by connecting dendrimer self-assembly and covalent (CD-Dendrimer) preparation of capillary coating, diazoresin and dendrimers through ionic bond is fixed on the inner wall of the capillary, through ultraviolet light irradiation, ionic bonds into covalent bonds, and fixed in a stable the inner wall of the capillary, the coating to separate the mixture of proteins, showed a good separation effect of.2. self-assembly (LBL) technique, based on the preparation of diazoresin and oxidation of Shi Moxi (GO) the capillary coating on the inner wall of the capillary, many two molecules alternate coatings, finally under UV irradiation, ionic bond the weak interaction between the original diazoresin and graphene oxide into covalent bonds more firmly, thereby preparing a covalent bond with controllable structure of multilayer composite coating, used for the separation of proteins, The ideal separation effect, reproducibility and stability is also very good.3. selection of hydrophilic substances and beta cyclodextrin (beta -CD) as protein material, resin and diazo through ionic bond in the capillary inner surface to form a composite film, coated capillary action, and then placed in UV irradiation, through optical the reaction of diazo resin is unique, ionic bonds into stable covalent bond, capillary coating is more stable, not easy to fall off, the adsorption inhibition of protein significantly, the covalent bond of prepared composite coating DR/ beta -CD capillary coating for the separation of protein mixture, the effect is very good. This thesis studies and capillary coating system the preparation is a new kind of coupling agent and the new combination of material, a new capillary coating prepared, not only the use of the advantages of the two kinds of coating technology, also some limitations made many improvements, non operation It is simple and fast. The coating is also very stable. It is used to separate the mixed protein to test its performance. The separation result is very good. The other test results are also good. It shows that the capillary coating prepared by us is stable and effective, which provides more possibilities for the further development of capillary.

【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：O658.9

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