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

doi:10.7503/cjcu20170742

Abstract

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

CLC Number:

O631.1⁺1

TrendMD:

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

Figures/Tables 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)

References 37

[1]	Christman K. L., Enriquez-Rios V. D., Maynard H. D., Soft Matter, 2006, 2(11), 928—939
[2]	Liu W., Li Y., Wang T., Li D., Fang L., Zhu S., Shen H., Zhang J., Sun H., Yang B., ACS Appl. Mater. Inter., 2013, 5(23), 12587—12593
[3]	Shen Y., Liu Y., Zhu G., Fang H., Huang Y., Jiang X., Wang Z.L., Nanoscale, 2013, 5(2), 527—531
[4]	Yao X., Peng R., Ding J., Advanced Materials, 2013, 25(37), 5257—5286
[5]	Shin D. S., Lee K. N., Janga K. H., Kim J. K., Chung W. J., Kim Y. K., Lee Y. S., Biosens. Bioelectron., 2003, 19(5), 485—494
[6]	Muguruma H., Takahashi H., Surf. Coat. Tech., 2010, 205(7), 2490—2494
[7]	Renault J. P., Bernard A., Bietsch A., Michel B., Bosshard H. R., Delamarche E., Kreiter M., Hecht B., Wild U. P., J. Phys. Chem. B, 2003, 107(3), 703—711
[8]	Alberto S. D. L., Campo A. D., Fernándezgarcía M., Rodríguezhernández J., Muñozbonilla A., ACS Appl. Mater. Inter., 2013, 5(9), 3943—3951
[9]	Chang-Yen D. A., Myszka D. G., Gale B. K., J. Microelectromech. Syst., 2006, 15(5), 1145—1151
[10]	Zhang A. J., Bai H., Li L., Chem. Rev., 2015, 115(18), 9801—9868
[11]	Bunz U .H. F., Adv. Mater., 2006, 18(8), 973—989
[12]	Sun W., Zhou Y. C., Chen Z. R., Acta Polym. Sin., 2012, (12), 1459—1464
	(孙巍, 周雨辰, 陈忠仁. 高分子学报, 2012, (12), 1459—1464)
[13]	Sun W., Chen Z .R., Chin. Polym. Bull., 2012, 8, 32—43
	(孙巍, 陈忠仁. 高分子通报, 2012, 8, 32—43)
[14]	Amirkhani M., Berger N., Abdelmohsen M., Zocholl F., Gonçalves M. R., Marti O. J., Polym. Sci. Pol. Phys., 2011, 49(20), 1430—1436
[15]	Lakshmi V., Raju A., Resmi V. G., Pancrecious J. ., Rajan T. P. D., Pavithran C., Phys. Chem. Chem. Phys., 2016, 18(10), 7367—7373
[16]	Sun W., Shao Z., Ji J., Polymer, 2010, 51(18), 4169—4175
[17]	Sun W., Zhou Y., Chen Z., Macromol. Res., 2012, 21(4), 414—418
[18]	Bu W. F., Li H. L., Sun H., Yin S. Y., Wu L. X., J. Am. Chem. Soc., 2005, 127(22), 8016—8017
[19]	Zhang Y., Wang C., Adv. Mater., 2007, 19(7), 913—916
[20]	Min E., Wong K. H., Stenzel M. H., Adv. Mater., 2008, 20(18), 3550—3556
[21]	Ju Y. L., Zhang Z. Z., Liu Y., Sun W., Chen Z.R., New Chem. Mater., 2014, 43(5), 89—92
	(鞠远来, 张震震, 刘玉, 孙巍, 陈忠仁. 化工新型材料, 2014, 43(5), 89—92)
[22]	Ding J., Zhang A., Bai H., Li L., Li J., Ma Z., Soft Matter, 2013, 9(2), 506—514
[23]	Zhang A., Du C., Bai H., Wang Y., Wang J., Li L., ACS Appl. Mater. Inter., 2014, 6(11), 89218927
[24]	Bormashenko E., Pogreb R., Stanevsky O., Bormashenko Y., Stein T., Gaisin V. Z., Cohen R., Gendelman O. V., Macromol. Mater. Eng., 2005, 290(2), 114—121
[25]	Muñozbonilla A., Ibarboure E., Papon E., Rodriguezhernandez J., Langmuir, 2009, 25(11), 6493—6499
[26]	Ma Y.Y., Liang J., Sun H., Wu L. X., Dang Y. Q., Wu Y. Q., Chem-Eur. J., 2012, 18(2), 526—531
[27]	Liang J., Ma Y., Sun H., Li W., Wu L., J. Colloid. Interf. Sci., 2013, 409(11), 80—87
[28]	Ma Y. Y., Zhou M. C., Walter S., Liang J., Chen Z. J., Wu L. X., Chem. Commun., 2014, 50(100), 15882—15885
[29]	Binks B .P., Curr. Opin. Colloid. In., 2002, 7(1/2), 21—41
[30]	Böker A., He J., Emrick T., Russell T.P., Soft Matter, 2007, 3(10), 1231—1248
[31]	Horozov T. S., Binks B. P., Angew. Chem. Int. Ed., 2006, 45(5), 773—776
[32]	Yang P. H., Sun W., Hu S., Chen Z. R., Prog. Chem., 2014, 26(7), 1107—1119
	(杨平辉, 孙巍, 胡思, 陈忠仁. 化学进展, 2014, 26(7), 1107—1119)
[33]	Sun W., Ji J., Shen J.C., Langmuir, 2008, 24(20), 11338—11341
[34]	Sun W., Zhou Y., Ju Y., Yang L., Xu T., Chen Z., Macromol. Chem. Phys., 2014, 215(1), 96—102
[35]	Yang P. H., Zhu J. F., Sun W., Zhou W. R., Chem. J. Chinese Universities, 2017, 38(4), 678—685
	(杨平辉, 朱嘉峰, 孙巍, 周婉容. 高等学校化学学报, 2017, 38(4), 678—685)
[36]	Hang J. J., Zhang Y. Q., Yang M. L., Sun W., Polym. Mater. Sci. Eng., 2017, 33(5), 142—146
	(黄俊杰, 张尹桥, 杨曼玲, 孙巍. 高分子材料科学与工程, 2017, 33(5), 142—146)
[37]	Wei Z., Wang C., Zou S., Liu H., Tong Z., Polymer, 2012, 53(6), 1229—1235