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过程工程学报 ›› 2015, Vol. 15 ›› Issue (3): 482-488.DOI: 10.12034/j.issn.1009-606X.215149

• 生化工程专栏 • 上一篇    下一篇

聚二乙烯基苯微球介质制备及其在奥曲肽层析分离中的应用

张晨旭 李强1 黄永东 赵岚 张荣月 巩方玲 宋翠 苏志国 袁其朋 马光辉   

  1. 北京化工大学生命科学与技术学院 中国科学院过程工程研究所生化工程国家重点实验室 中国科学院过程工程研究所生化工程国家重点实验室 中国科学院过程工程研究所生化工程国家重点实验室 中国科学院过程工程研究所生化工程国家重点实验室 中国科学院过程工程研究所生化工程国家重点实验室 中国科学院过程工程研究所生化工程国家重点实验室 中国科学院过程工程研究所生化工程国家重点实验室 北京化工大学生命科学与技术学院 中国科学院过程工程研究所生化工程国家重点实验室
  • 收稿日期:2015-03-04 修回日期:2015-04-14 出版日期:2015-06-20 发布日期:2015-06-20
  • 通讯作者: 张晨旭

Chromatographic Separation of Octreotide with Prepared Polydivinylbenzene Microspheres

ZHANG Chen-xu LI Qiang HUANG Yong-dong, ZHAO Lan ZHANG Rong-yue GONG Fang-ling, SONG Cui SU Zhi-guo YUAN Qi-peng MA Guang-hui   

  1. College of Life Science and Technology, Beijing University of Chemical Technology National Key Lab of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences State Key laboratory of Biochemical Engineering, Institute of Process Engineering, CAS State Key Lab.Biochem.Eng.,Institute of Process Engineering, Chinese Academy of Sciences State Key Lab.Biochem.Eng.,Institute of Process Engineering, Chinese Academy of Sciences State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences State Key Lab. Biochem. Eng., Inst. Process Eng., Chinese Academy of Sciences School of Life Science and Technology, Beijing University of Chemical Technology State Key Lab. Biochem. Eng., Inst. Process Eng., Chinese Academy of Sciences
  • Received:2015-03-04 Revised:2015-04-14 Online:2015-06-20 Published:2015-06-20
  • Contact: ZHANG Chen-xu

摘要: 以二乙烯基苯为单体,采用微孔膜乳化法制备了尺寸均一的水包油乳液,在75℃下悬浮聚合20 h,制得均一的聚二乙烯基苯微球填料,粒径为14.2 mm,粒径分布系数Rspan=0.71,孔径为65 nm,比表面积为160 m2/g. 通过高压匀浆法将合成的微球介质高效装填不锈钢液相色谱柱(f4.6 mm′250 mm),使柱效达11000塔板数. 利用装填的色谱柱进行多肽药物奥曲肽的分离纯化,奥曲肽从固相合成粗品中快速有效分离,纯度由42.89%提高到99.99%.

关键词: 奥曲肽, 反相介质, 聚二乙烯基苯, 微球, 层析, 纯化

Abstract: Polydivinylbenzene (pDVB) microspheres with the particle size of 14.2 mm were prepared with divinylbenzene as monomer by membrane emulsification and suspension polymerization at 75℃ for 20 h. The prepared pDVB microspheres had uniform particle size (Rspan=0.71), mean pore size of 65 nm and specific surface area of 160 m2/g. The pDVB microspheres were packed into f4.6 mm′250 mm stainless steel column by slurry packing method, and tested in separation of peptide drug by high performance liquid chromatography. Afterword, a very fast separation process of octreotide from synthetic peptide crude mixtures on pDVB reversed phase was carried out. Fourier-transformed infrared resonance (FT-IR) spectroscopy, and reversed-phase HPLC combined with electrospray ionisation mass spectrometry (ESI-MS) were used for analysis of resultant peptides. Under the optimized conditions, the octreotide was purified from 42.89% to 99.99%.

Key words: octreotide, reversed phase media, polydivinylbenzene, microspheres, chromatography, purification

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