高压静电纺丝法制备分离纯化木瓜蛋白酶的亲和膜基材的应用研究

发布时间：2018-05-11 08:11

本文选题：亲和膜色谱 + 静电纺丝　；参考：《东华大学》2010年硕士论文

【摘要】： 近年来,亲和膜色谱技术在蛋白质的分离纯化中得到了快速发展,与柱亲和色谱相比,亲和色谱具有许多优点,如流速高、分离速度快、压降低和易于规模化生产等。利用高压静电纺丝技术制造的具有纳米或微米尺度的聚合物复合纤维膜是作为亲和膜色谱基材的良好原料。本课题就从选择理想亲和膜基材的角度出发,成功地利用高压静电纺丝技术制备出不同混合比例的尼龙6/壳聚糖纳米复合纤维膜。首先介绍了静电纺丝的发展、方法、成丝理论、成丝影响因素以及静电纺纳米纤维的广阔应用前景。通过实验为尼龙6/壳聚糖静电纺丝找到了合适的溶剂体系,即体积比为9/1的六氟异丙醇/甲酸混合体系,首次制备出了尼龙6/壳聚糖静电纺纳米纤维膜。研究了一些纺丝参数(纺丝电压、给液速率、纺丝距离),纺丝液浓度和尼龙6/壳聚糖不同比例配比对纺丝结果的影响。对不同比例的尼龙6/壳聚糖共混静电纺纳米纤维的形态、性能进行了研究。通过扫描电镜分析发现制备的静电纺纳米纤维直径分布在80-310 nm范围内;通过红外光谱分析和DSC分析,发现尼龙6和壳聚糖分子之间存在一定的相互作用;X射线衍射分析证明,在经过静电纺丝以后,纤维的物相结构发生了变化,静电纺丝纤维更趋向无定型态;通过对电纺纤维的接触角测试和力学性能显示,所得的复合纤维具有良好的亲水性能以及良好的机械性能。本研究的首要目的是制备出具有更高蛋白质吸附能力的新型亲和膜材料。本论文首次以静电纺尼龙6/壳聚糖为材料,以活性染料Cibacron Blue F3GA为配基制备了一种新型的分离木瓜蛋白酶的亲和膜基材,研究了其物理性能和色谱性能。实验结果表明,染料F3GA的键和量为102μmol/g,亲和膜对木瓜蛋白酶(2.0 mg/ml)的饱和吸附量达到93.46 mg/g,利用该亲和膜对木瓜蛋白酶原液的纯化倍数提高了5倍。本文综合运用批量法和动态法以亲和膜对木瓜蛋白酶的吸附过程进行研究,结果表明,亲和膜对木瓜蛋白酶的吸附行为满足Langmuir模型。综上,本研究所制备以静电纺尼龙6/壳聚糖纳米复合纤维膜为基材,以活性染料F3GA为配基的亲和膜体系,适合规模化分离纯化木瓜蛋白酶。
[Abstract]:In recent years, affinity membrane chromatography technology has been rapidly developed in the separation and purification of proteins. Compared with column affinity chromatography, affinity chromatography has many advantages, such as high flow rate, fast separation speed, low pressure and easy to scale production. Polymer composite fiber membrane with nanometer or micron scale produced by high voltage electrostatic spinning technology is a good raw material for affinity membrane chromatography. In this paper, the nylon 6 / chitosan nanocomposite fiber membrane with different mixing ratio was successfully prepared by high voltage electrostatic spinning from the point of view of choosing ideal affinity membrane substrate. In this paper, the development, method, theory of electrospinning, the influence factors of electrospinning and the wide application prospect of electrospun nanofibers are introduced. The proper solvent system for the electrospinning of nylon 6 / chitosan, that is, hexafluoroisopropanol / formic acid mixed system with volume ratio of 9 / 1, was found through experiments, and nylon 6 / chitosan nanofiber film was prepared for the first time. The effects of spinning parameters (spinning voltage, feed rate, spinning distance), concentration of spinning solution and ratio of nylon 6 / chitosan on spinning results were studied. The morphology and properties of nylon 6 / chitosan blended nanofibers were studied. The diameter of the electrospun nanofibers was found to be in the range of 80-310 nm by SEM analysis, and by IR and DSC analysis, it was found that there was a certain interaction between nylon 6 and chitosan molecules. After electrospinning, the phase structure of the fiber changes, and the electrostatic spinning fiber tends to be amorphous. The contact angle of the electrospun fiber is measured and the mechanical properties are shown. The obtained composite fiber has good hydrophilic and mechanical properties. The primary aim of this study is to prepare new affinity membrane materials with higher protein adsorption capacity. In this paper, a novel affinity membrane substrate for the separation of papain was prepared from electrospun nylon 6 / chitosan with reactive dye Cibacron Blue F3GA as ligand for the first time, and its physical and chromatographic properties were studied. The results showed that the bond sum of dye F3GA was 102 渭 mol / g, and the saturated adsorption capacity of the affinity membrane for papain was 93.46 mg / g, and the purification multiple of papain was increased by 5 times. The adsorption process of papain by affinity membrane was studied by batch method and dynamic method. The results showed that the adsorption behavior of papain on affinity membrane satisfied Langmuir model. In conclusion, an affinity membrane system was prepared using electrospun nylon 6 / chitosan nanocomposite fiber membrane as substrate and reactive dye F3GA as ligand, which was suitable for large-scale separation and purification of papain.
【学位授予单位】：东华大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：TS104.7

【相似文献】