复乳法制备大孔琼脂糖分离介质与疫苗结合性能研究

摘要大孔分离介质在分离纯化特别是超大生物分子分离纯化领域具有十分重要的应用意义。受材料性质和制备技术所限制，大孔分离介质以聚合物材料最为常见，针对琼脂糖的大孔分离介质较为少见。通过复乳法制备大孔琼脂糖微球，研究制备条件对微球形貌与成孔情况以及粒径与分布等的影响，发现该微球具备微米级超大孔结构，且制备条件对孔道结构的影响呈现一定规律，琼脂糖溶液浓度、内/外油相与水相比例、初乳转速和成球转速等都对成孔性至关重要。激光共聚焦显微镜实验结果表明，衍生后的大孔琼脂糖分离介质对乙肝表面抗原具有良好的结合能力，使其能够完全进入微球内部。与传统琼脂糖介质相比，大孔琼脂糖介质能够更快速、充分结合大分子疫苗，在超大分子分离纯化领域应用前景广阔。

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	黄兰
	黄永东
	赵岚
	朱凯
	吴学星
	周丹妮
	苏志国
	马光辉
	王启宝

关键词：大孔琼脂糖微球；复乳法；致孔；离子交换介质；乙肝表面抗原

Abstract： The macroporous chromatographic media are of great importance in the field of bioseparation, especially for large biomolecules. Limited by both of the material properties and preparation technology, the macroporous chromatographic media were mostly prepared from synthetic polymers, and native polymers, e.g., agarose, were rarely used. In this paper, macroporous agarose microspheres were prepared by double emulsification method. The effects of preparation conditions on microsphere morphology and pore formation, as well as particle size and distribution were studied. It was found that the microspheres have micro-scale super-large pore structure and the pore structure shows certain regularity due to the influence of preparation conditions. The concentration of agarose solution, the ratio of internal/external oil phase to water, the speed of colostrum and the speed of ball formation are all important for pore formation. Confocal laser scanning microscopy results showed that the macroporous agarose microspheres after being functionalized with ion-exchange groups had a good binding ability to Hepatitis B Surface antigen (HBsAg) and this antigen can completely enter the inside of the microspheres. Compared with conventional agarose-based chromatographic media, macroporous agarose-based media combined macromolecular vaccine more quickly and fully, and had wide prospects of application in the field of purification of large biomolecules.

Keywords： macroporous agarose-based microspheres； double emulsification method； pore-forming； ion-exchange chromatographic media； HBsAg

Received 2019-02-28;

Fund:国家重点研发计划资助项目（2018YFC0311101；2016YFF0202304）；国家自然科学基金资助项目（21476241）；中国科学院重点部署项目（KFZD-SW-218）；北京市自然科学基金资助项目（2172054）。

Corresponding Authors: 黄永东,E-mail:ydhuang@ipe.ac.cn;王启宝,E-mail:wqb@cumtb.edu.cn。 Email: ydhuang@ipe.ac.cn;wqb@cumtb.edu.cn

About author: 黄兰(1994-),女,硕士研究生,从事化学工艺方向研究。

引用本文:

黄兰, 黄永东, 赵岚, 朱凯, 吴学星, 周丹妮, 苏志国, 马光辉, 王启宝.复乳法制备大孔琼脂糖分离介质与疫苗结合性能研究[J]. 化学工业与工程, 2019,36(4): 70-79

Huang Lan, Huang Yongdong, Zhao Lan, Zhu Kai, Wu Xuexing, Zhou Danni, Su Zhiguo, Ma Guanghui, Wang Qibao.Preparation of Macroporous Agarose-Based Chromatographic Media Using a Double Emulsification Method and Its Binding with HBsAg[J]. Chemcial Industry and Engineering, 2019,36(4): 70-79

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