本文選題：醋酸纖維素　切入點(diǎn)：親和膜色譜　出處：《北京化工大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：隨著社會(huì)對(duì)生物活性大分子的需求越來越高,很多領(lǐng)域如醫(yī)藥、生物等對(duì)分離技術(shù)的純度要求也越來越高。膜色譜技術(shù)被認(rèn)為是一種高效節(jié)能環(huán)保的分離技術(shù),且具有易裝填和放大的優(yōu)點(diǎn),工業(yè)化前景很好。本研究旨在制備一種親和膜色譜,通過在膜上接枝適當(dāng)?shù)呐浠杂H和吸附相應(yīng)的生物大分子,以達(dá)到分離純化該物質(zhì)的目的。首先,本文探索了相轉(zhuǎn)化法制備醋酸纖維素膜色譜的可行性。通過優(yōu)化濃度、溶劑配比、添加劑、蒸氣浴濕度與溫度等條件,醋酸纖維素膜的純水通量從283 Lm-2h-1提高到1598Lm-2h-1,相同方法制得的醋酸纖維素接枝p-環(huán)糊精膜純水通量為639 Lm-2h-1,但該接枝膜填裝膜色譜時(shí)最多僅可添加3層,且隨著壓力增大膜內(nèi)部孔道被完全壓塌,因此認(rèn)為相轉(zhuǎn)化法制得的醋酸纖維素膜不適合作為親和膜色譜的底膜使用。其次,本文嘗試用靜電紡絲法制備醋酸纖維素膜,該膜具有較高的孔隙率和機(jī)械性能,通過優(yōu)化濃度、溶劑配比、接收距離、流速、電壓等條件制備出的醋酸纖維素膜纖維直徑在264nm到5.121μm之間,且具有良好形態(tài)。醋酸纖維素膜去乙�；蠼又-環(huán)糊精所制得的接枝物膜可裝填25層膜色譜且連續(xù)使用24h,葛根素粗品經(jīng)過該膜色譜處理后,葛根素純度由42.52%提高到84.1%,由此可證明靜電紡絲法制得的醋酸纖維素膜可作為膜色譜底膜使用。最后,本文制備了可特異性識(shí)別蛋白SH3結(jié)構(gòu)域的小分子多肽LPPLPLPPLP,將靜電紡絲法制得的醋酸纖維素膜經(jīng)去乙酰化、氧化處理后接枝小分子多肽LPPLPLPPLP,接枝率可達(dá)15.1μg/mg,把BSA做雜質(zhì)與磷脂酶A2混合成磷脂酶粗品,考察膜色譜分離性能。結(jié)果表明,將接枝小分子多肽LPPLPLPPLP的親和膜填裝成20層膜色譜,粗品溶液經(jīng)過該膜色譜的處理,可有效將磷脂酶A2固定在膜色譜上,飽和吸附率為7.2μg/mg,再通過收集膜色譜洗脫液并濃縮,由此達(dá)到純化磷脂酶A2的目的。
[Abstract]:With the increasing demand for bioactive macromolecules in society, many fields, such as medicine, biology and so on, require higher purity of separation technology. Membrane chromatography technology is considered to be a kind of high efficiency, energy saving and environmental protection separation technology. It has the advantages of easy loading and amplification, and has a good prospect of industrialization. The aim of this study was to prepare an affinity membrane chromatography, and to adsorb the corresponding biomolecules by grafting appropriate ligand on the membrane. First of all, the feasibility of preparing cellulose acetate membrane chromatography by phase inversion method was explored. By optimizing concentration, solvent ratio, additive, vapor bath humidity and temperature, etc. The pure water flux of cellulose acetate membrane was increased from 283Lm-2h-1 to 1598Lm-2h-1.The pure water flux of cellulose acetate grafted with p-cyclodextrin was 639Lm-2h-1, but only three layers could be added in the filling membrane chromatography. With the increase of pressure, the inner pore of the membrane is completely collapsed, so it is considered that the cellulose acetate membrane prepared by phase transformation method is not suitable for use as the bottom membrane of affinity membrane chromatography. Secondly, this paper attempts to prepare cellulose acetate membrane by electrostatic spinning method. The film has high porosity and mechanical properties. The fiber diameter of cellulose acetate membrane is between 264 nm and 5.121 渭 m by optimizing concentration, solvent ratio, receiving distance, flow rate, voltage and so on. After deacetylation of cellulose acetate membrane, the graft film prepared by grafting pcyclodextrin can be loaded with 25 layers of membrane chromatography and used continuously for 24 hours. The crude product of puerarin can be treated by the membrane chromatography. The purity of puerarin increased from 42.52% to 84.1, which proved that the cellulose acetate membrane prepared by electrospinning can be used as the bottom film of membrane chromatography. In this paper, a small molecular peptide, LPPLPLPPLP, which can specifically recognize the domain of protein SH3, was prepared. The cellulose acetate membrane prepared by electrospinning was deacetylated. After oxidation treatment, the graft rate of LPPLPLPPLP was 15.1 渭 g / mg. BSA was mixed with phospholipase A2 to form phospholipase crude product. The results showed that the affinity membrane of LPPLPLPPLP was filled into a 20-layer membrane chromatography. The crude solution was treated by the membrane chromatography and phospholipase A2 was effectively immobilized on the membrane chromatography. The saturated adsorption rate was 7.2 渭 g / mg. then the phospholipase A2 was purified by collecting the eluent and concentrating it.
【學(xué)位授予單位】：北京化工大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：TQ051.893;O652.63

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本文編號(hào)：1599835