利用反相乳液-水滴模板法制备功能性生物基图案化结构

doi:10.7503/cjcu20170742

高等学校化学学报 ›› 2018, Vol. 39 ›› Issue (6): 1311.doi: 10.7503/cjcu20170742

利用反相乳液-水滴模板法制备功能性生物基图案化结构

丁凌云, 鞠远来, 孙巍(), 陈吉

宁波大学材料科学与化学工程学院, 宁波市特种高分子材料制备与应用技术重点实验室, 宁波 315211

收稿日期:2017-11-17 出版日期:2018-06-10 发布日期:2018-04-12
基金资助:
国家自然科学基金(批准号: 21104036)、浙江省重点科技创新团队自设项目(批准号: 2011R50001-04)、宁波市自然科学基金(批准号: 2015A610057)和宁波大学王宽诚幸福基金资助.

Preparation of Functional Patterned Protein Arrays via the Combination of Inverse Emulsion and Breath Figure Method^†

DING Lingyun, JU Yuanlai, SUN Wei^*(), CHEN Ji

Department of Materials Science and Engineering, Key Laboratory of Specialty Polymers, Faculty of Materials Science and Chemical Engineering, Ningbo University, Ningbo 315211, China

Received:2017-11-17 Online:2018-06-10 Published:2018-04-12
Contact: SUN Wei E-mail:sunwei@nbu.edu.cn
Supported by:
† Supported by the National Natural Science Foundation of China(No.21104036), the Program for Zhejiang Leading S&T Innovation Team, China(No.2011R50001-04), the Natural Science Foundation of Ningbo, China(No.2015A610057) and the K. C. Wong Magna Fund in Ningbo University, China.

摘要/Abstract

摘要：

通过将传统水滴模板法中的单相成膜液改造为反相乳液体系, 引进乳液水滴来加载蛋白质组分, 实现了对亲水组分直接实施水滴模板法从而获得其阵列组装结构. 利用壳聚糖纳米粒子作为皮克林反相乳液的乳化剂制备了稳定的反相乳液体系, 并进一步得到聚乳酸/壳聚糖/牛血清白蛋白的杂化蜂窝状多孔薄膜, 考察了壳聚糖粒子对于乳液稳定和所制备多孔阵列结构形貌的影响, 研究了加入蛋白质浓度和乳液中的水相/油相比例对于所成膜孔形貌的影响, 利用荧光标记蛋白质跟踪确认基于蜂窝状多孔阵列结构上的图案化蛋白质阵列组装形貌.

关键词: 壳聚糖粒子, 反相乳液, 水滴模板法, 蛋白质阵列结构, 蜂窝状多孔表面

Abstract:

We established a modified breath figure(BF) method using inverse emulsion as casting solution to achieve directed assembly of water-soluble components with the BF arrays. It is a one-step technique with great ease of fabricating regular arrays of biomolecules via self-assembly. To be specific, we constructed a hybrid structured film made from polylactic acid(PLA)/chitosan(CS)/bovine serum albumin(BSA) mixing emulsion. CS particles were synthesized to serve as stabilizers for the inverse emulsion of water-in-oil type and BSA was selected as model protein to be carried within the emulsion water droplets. It has been proved that the CS particles could not only effectively contribute to stabilize the inverse emulsion, but also be able to improve the regularity of the obtained BF arrays. Furthermore, the influencing factors of BSA concentration and water-to-oil ratio were carefully tuned to study their effect on the obtained surface morphology. The results showed that the increase of the BSA concentration and the water-to-oil ratio would both compromise the regular arrangement of the surface pore arrays. But if the concentration and ratio were both increased to a high level, the regular BF arrays would reappear. We tagged the BSA with fluorescent probe to track the assembled morphology of the BSA within the porous polymer matrix. And 3D ring-like arrays of the fluorescent protein were observed under confocal microscope, indicating the success of obtaining the protein arrays via a one-step technique.

Key words: Chitosan particle, Inverse emulsion, Breath figure method, Protein array, Honeycomb-structured porous surface

中图分类号:

O631.1⁺1

TrendMD:

丁凌云, 鞠远来, 孙巍, 陈吉. 利用反相乳液-水滴模板法制备功能性生物基图案化结构. 高等学校化学学报, 2018, 39(6): 1311.

DING Lingyun, JU Yuanlai, SUN Wei, CHEN Ji. Preparation of Functional Patterned Protein Arrays via the Combination of Inverse Emulsion and Breath Figure Method^†. Chem. J. Chinese Universities, 2018, 39(6): 1311.

图/表 6

Fig.1 Size distribution of CS particles determined by the DLS method(A) and TEM image of CS particles(B)

Fig.2 Optical micrographs of the honeycomb-patterned film prepared from the inverse solutions at different volume ratios of water phase without(A—D) or with chitosan particles(A'—D')Volume ratio of water/oil: (A, A') 1%; (B, B') 2%; (C, C') 3%; (D, D') 5%.

Fig.3 SEM images of the top-view(A) and cross-view(B) of porous films fabricated from inverse emulsion containing chitosan particles with the water/oil ratio of 1%

Fig.4 Optical micrographs of the honeycomb-patterned films prepared from the inverse emulsions at different volume ratios of water phase with different BSA concentrationsBSA concentration/(mg·mL-1): (A1—E1) 2.5; (A2—E2) 5.0; (A3—E3) 7.5; (A4—E4) 10.0. Volume ratio of water/oil: (A1—A4) 1%; (B1—B4) 2%; (C1—C4) 3%; (D1—D4) 4%; (E1—E4) 5%.

Fig.5 Top-view(A, C) and cross-sectional(B, D) SEM images of the honeycomb-patterned film prepared from the inverse emulsions at volume ratios of 1%(protein concentration of 5 mg/mL)(A,B) and 5%(protein concentration of 10 mg/mL)(C,D)

Fig.6 Top sectional(A) and 3D reconstructed(B) CLSM images of the honeycomb-patterned film prepared from the inverse solutions(2% BSA aqueous solution with BSA concentration of 10 mg/mL)

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利用反相乳液-水滴模板法制备功能性生物基图案化结构

Preparation of Functional Patterned Protein Arrays via the Combination of Inverse Emulsion and Breath Figure Method^†

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利用反相乳液-水滴模板法制备功能性生物基图案化结构

Preparation of Functional Patterned Protein Arrays via the Combination of Inverse Emulsion and Breath Figure Method†

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Preparation of Functional Patterned Protein Arrays via the Combination of Inverse Emulsion and Breath Figure Method^†