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题名	均一聚（二乙烯基苯）多孔微球的制备及在多肽分离纯化中的应用
作者	王静
学位类别	硕士
答辩日期	2009-06-02
授予单位	中国科学院过程工程研究所
授予地点	过程工程研究所
导师	马光辉
关键词	聚（二乙烯基苯）微球 聚合物填料 高效液相色谱 多肽 分离纯化
其他题名	Preparation of Uniform Porous Polydivinylbenzene Microspheres and Their Applications in Separation and Purification of Peptides
学位专业	生物化工
中文摘要	在本研究中，我们通过微孔膜乳化-悬浮聚合法制备了尺寸均一、粒径和孔径均可控、且成本低、可规模化生产的聚（二乙烯基苯）（Polydivinylbenzene, PDVB）微球，并将其用于多肽的分离纯化。论文分为三个部分： 第一部分采用微孔膜乳化-悬浮聚合法，在实验室前期工作的基础上，通过对影响微球粒径大小及分布的因素进一步考察，制备出了粒径均一、比表面积高、孔径范围适合多肽分离的PDVB微球。 第二部分为通过对微球机械强度的考察和匀浆剂、顶替液的选择，确定了适合PDVB微球的装柱条件，制备了性能优良的300 mm×10 mm I.D.不锈钢半制备柱，并将其分别用于苯同系物、酚类、多肽类混合物的分离，达到了良好的分离效果，相邻各峰间分离度均大于1.5，谱图峰形对称，酚类、多肽的柱效分别达13000 m-1和21000 m-1。 第三部分使用PDVB微球装填的半制备柱，通过一步反相液相色谱分离，对4种合成多肽分别进行了成功的纯化。其中，粒径为30 μm的PDVB介质可将15肽纯度由38.1%提高到78.6%；30 μm和25 μm的PDVB介质分别可将18肽纯度由35.7%提高到98.47%和99.85%，对应的回收率分别为24.44%和64.14%，色谱柱分辨率随介质粒径减小提高显著；25 μm的PDVB介质可将16肽纯度由54.0%提高到96.83%；强疏水性的24肽在C18柱上无法洗脱，在自制PDVB半制备柱上洗脱的峰形与SOURCE商品柱相似，优于C4商品柱。
英文摘要	In this work, uniform size and pore controllable, low cost and large-scale-productive polydivinylbenzene (PDVB）microspheres were prepared by membrane emulsification technology, then were used in purification of peptides. This work was carried out from the following 3 parts. In the first part, PDVB microspheres with uniform size, high specific surface area and pore-size suitable for peptides purification were prepared by membrane emulsification technology after optimizing the factors affecting particle size and particle distribution. In the second part, high performance semi-preparative stainless steel columns (300 mm×10 mm I.D.) were obtained by optimization of pressure and solvents. Then the columns were used to separate the mixture of benzene and its derivatives, phenols and peptides, and all showed high performance. Resolution of adjacent peaks was higher than 1.5, spectrum peak shape was symmetrical and column efficiency of phenols and peptides reached 13000 m-1 and 21000 m-1, respectively. In the third par, four solid phase synthetic peptides were successfully purified respectively by one-step RP-HPLC using the semi-preparative columns. Purity of 15-peptide was improved from 38.1% to 78.6% with 30 μm PDVB microspheres; purities of 18-peptide was improved from 35.7% to 99.85% and 98.47% with 25 μm and 30 μm PDVB microspheres respectively, and the corresponding recoveries were 64.14% and 24.44%, which showed resolution of columns increased remarkably as particle sizes of packings decreased; purity of 16-peptide was improved from 54.0% to 96.83% with 25 μm PDVB microspheres; 24-peptide can’t be eluted with C18 columns due to its strong hydrophobic property, but the shape of elution peak with PDVB semi-preparative column was similar with commercial SOURCE column and better than commercial C4 column.
语种	中文
公开日期	2013-09-16
页码	95
内容类型	学位论文
源URL	[http://ir.ipe.ac.cn/handle/122111/1295]
专题	过程工程研究所_研究所（批量导入）
推荐引用方式 GB/T 7714	王静. 均一聚（二乙烯基苯）多孔微球的制备及在多肽分离纯化中的应用[D]. 过程工程研究所. 中国科学院过程工程研究所. 2009.

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