本文選題：再生纖維素膜　切入點：固定化金屬離子親和膜　出處：《西北大學(xué)》2015年碩士論文

【摘要】：固定金屬親和膜色譜具有可以在較短時間內(nèi)處理大量的實際樣品,低壓降,容易擴(kuò)大化生產(chǎn)等優(yōu)點,因此在蛋白等生物大分子的分離純化方面有潛在應(yīng)用價值。但是與傳統(tǒng)的以微球裝柱的柱色譜相比,膜色譜比表面積較低,因而對蛋白的吸附容量較低,限制其分離效率的提高。為提高蛋白的吸附量,本文采用超支化方法對膜進(jìn)行表面改性,通過在表面引入更多的功能基團(tuán),增加吸附位點來提高蛋白的吸附量,制備了一種對于金屬離子和蛋白都有較高吸附量的超支化固定金屬離子親和膜。同時研究了該改性膜對蛋白的吸附性能,并將其用于含組氨酸標(biāo)簽蛋白的分離純化。首先通過兩步反應(yīng)在膜表面引入帶有氨基的宮能團(tuán),接下來將丙烯酸甲酯和乙二胺逐步加入,通過邁克加成反應(yīng)和酰胺化反應(yīng)在膜表面形成超支化結(jié)構(gòu),最后依次與環(huán)氧氯丙烷和亞氨基二乙酸反應(yīng),再螯合金屬離子制備了超支化固定金屬離子親和膜。用FTIR和XPS表征了膜表面的化學(xué)組成。溶菌酶被作為模型蛋白來探究蛋白的吸附性能,該改性膜對于溶菌酶的最大吸附量為162 mg/cm3,高于文獻(xiàn)報道值。同時探究了溶液pH,離子強(qiáng)度,疏水作用和吸附時間對吸附量的影響。將該改性膜用于蛋清中溶菌酶的純化,回收率可達(dá)到94.3%,比未超支化的固定金屬離子親和膜的回收率提高了50%。最后用本文制備的改性膜從大腸桿菌粗提液中分離和富集含六聚組氨酸標(biāo)簽的天蠶抗菌肽B-人表皮生長因子(CB-EGF)融合蛋白,獲得的蛋白純度較高。所有這些結(jié)果證明本文合成的超支化固化金屬離子親和膜在His-tagged蛋白的分離純化方面有潛在的應(yīng)用價值。
[Abstract]:Stationary metal affinity membrane chromatography has the advantages of being able to deal with a large number of actual samples in a relatively short time, falling at low pressure, easy to expand production, etc. Therefore, it has potential application value in the separation and purification of biological macromolecules such as protein, but compared with the traditional column chromatography with microspheres, the specific surface area of membrane chromatography is lower, so the adsorption capacity of protein is lower. In order to improve the adsorption capacity of protein, the hyperbranched method was used to modify the surface of the membrane. By introducing more functional groups on the surface and increasing the adsorption sites, the adsorption capacity of the protein was increased. A hyperbranched fixed metal ion affinity membrane with high adsorption capacity for both metal ions and proteins was prepared. It was used for the separation and purification of histidine labelled proteins. Firstly, a hysteric group with amino groups was introduced on the membrane surface through two steps reaction, and then methyl acrylate and ethylenediamine were added step by step. The hyperbranched structure was formed on the membrane surface by Michael addition reaction and amidation reaction, and then reacted with epichlorohydrin and iminodiacetic acid. The hyperbranched immobilized metal ion affinity membrane was prepared by chelating metal ions. The chemical composition of the membrane surface was characterized by FTIR and XPS. Lysozyme was used as a model protein to investigate the adsorption properties of the membrane. The maximum adsorption capacity of the modified membrane for lysozyme was 162 mg / cm 3, which was higher than that reported in the literature. The effects of pH, ionic strength, hydrophobic action and adsorption time on the adsorption capacity of the modified membrane were investigated. The modified membrane was used to purify lysozyme from egg white. The recovery rate was 94. 3%, which was 50% higher than that of unhyperbranched immobilized metal ion affinity membrane. Finally, the modified membrane prepared in this paper was used to separate and enrich the antimicrobial peptide B- of Bombyx mori containing Hexahistidine label from the crude extract of Escherichia coli. Human epidermal growth factor CB-EGF fusion protein, All these results show that the hyperbranched solidified metal ion affinity membrane synthesized in this paper has potential application value in the separation and purification of His-tagged protein.
【學(xué)位授予單位】：西北大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：TQ051.893

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 柴紅,錢驊,陳歡林;親和膜分離應(yīng)用研究進(jìn)展[J];膜科學(xué)與技術(shù);2003年04期

2 聶華麗;陳天翔;朱利民;;尼龍親和膜的制備條件優(yōu)化及其表征[J];膜科學(xué)與技術(shù);2011年02期

3 鮑時翔,鄭青,姚汝華,蘇志國;親和膜吸附γ-免疫球蛋白的研究[J];華南理工大學(xué)學(xué)報(自然科學(xué)版);1998年01期

4 郭為,商振華,于億年,周康,周良模;大孔纖維素親和膜在生化制劑純化中的應(yīng)用[J];中國藥學(xué)雜志;1998年12期

5 李靜,陳歡林,柴紅;金屬螯合親和膜吸附分離與純化溶菌酶的研究(Ⅱ)──不同金屬螯合親和膜對溶菌酶的吸附性能[J];高等學(xué)�；瘜W(xué)學(xué)報;1999年08期

6 商振華,周冬梅,郭為,于億年,周良模,潘明臣,周康;聚酰胺親和膜用于去除內(nèi)毒素的研究 Ⅰ，

本文編號：1688635