3D打印便携式凝胶电泳装置用于蛋白质的快速检测

引用本文: 李莹莹, 王丁一, 农骐郢, 刘丽红, 张蒙, 梁勇, 胡立刚, 何滨, 江桂斌. 3D打印便携式凝胶电泳装置用于蛋白质的快速检测[J]. 色谱, 2020, 38(11): 1316-1322. doi: 10.3724/SP.J.1123.2020.02018 shu

Citation: LI Yingying, WANG Dingyi, NONG Qiying, LIU Lihong, ZHANG Meng, LIANG Yong, HU Ligang, HE Bin, JIANG Guibin. 3D printed portable gel electrophoresis device for rapid detection of proteins[J]. Chinese Journal of Chromatography, 2020, 38(11): 1316-1322. doi: 10.3724/SP.J.1123.2020.02018 shu

3D打印便携式凝胶电泳装置用于蛋白质的快速检测

李莹莹 ^1,2, ,
王丁一 ^2, ,
农骐郢 ^2, ,
刘丽红 ^2, ,
张蒙 ^1, ,
梁勇 ^1,3, ,
胡立刚 ^{1,2,
,} ,
何滨 ^2, ,
江桂斌 ^2,

1.
江汉大学环境与健康研究院, 湖北武汉 430056;
2.
中国科学院生态环境研究中心, 环境化学与生态毒理学国家重点实验室, 北京 100085;
3.
持久性有毒污染物环境与健康危害湖北省重点实验室, 湖北武汉 430056

通讯作者: 胡立刚, E-mail:lghu@rcees.ac.cn

基金项目:
国家自然科学基金（21577153，91743203）.

摘要: 随着现场分析对于快速、便携和经济型检测的需求，分析仪器的便携化和微型化备受关注。3D打印技术的不断发展，将会极大推动小型化、便携式实验设备的开发和研制。分析仪器的微型化有助于促进资源不足地区在医疗现场、食品安全和环境污染等方面的现场监测。目前，用于蛋白质分离的凝胶电泳装置多为实验室用小型化分析仪器，可用于现场快速分离蛋白质的小型化仪器尚未见报道。该研究设计加工了一款便携式凝胶电泳装置，用于蛋白质的快速分离检测。首先，通过3D打印加工的凝胶电泳装置可在实验室内方便、快捷、低成本的复制。其次，通过对预染蛋白质相对分子质量标准的分离测试，对该系统结构进行优化。优化后该凝胶电泳装置电泳槽的尺寸仅为15 mm×20 mm×17 mm，采用3D打印技术可在5 h内加工完成，耗费打印材料10 mL。正负极所用电泳缓冲液共需4 mL，所使用的25 V锂电池可实现100 h左右的工作时间。装置优化后可实现蛋白质的快速高效分离。随后，在5种常用蛋白质相对分子质量标准的分离中，该装置与商业化平板凝胶电泳分离效果相当，同时具备更快的分离速度。该研究在便携式凝胶电泳装置的开发及其在蛋白质快速分离方面取得了初步成果，但在分离完成后立即对蛋白质进行定量分析以及更多实际样品的应用方面还需要进一步研究。

关键词:

凝胶电泳
/ 3D打印
/ 蛋白质
/ 便携式
/ 分离

English

3D printed portable gel electrophoresis device for rapid detection of proteins

1.
Institute of Environment and Health, Jianghan University, Wuhan 430056, China;
2.
State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China;
3.
Hubei Key Laboratory of Environmental and Health Hazards of Persistent Toxic Substances, Wuhan 430056, China

Corresponding author: HU Ligang, lghu@rcees.ac.cn

Received Date: 26 February 2020
Fund Project:
National Natural Science Foundation of China (Nos. 21577153, 91743203).

Abstract: The growing demand for rapid, portable, and economical detection methods for environmental analysis has resulted in increasing demands on the portability and miniaturization of analytical instruments. The miniaturization of scientific instruments facilitates analysis in the field of medicine, food, and environment, especially for the under-resourced areas. The gel electrophoresis devices currently available for protein separation are primarily used in laboratories. Miniaturized instruments that can be used for on-site and rapid separation of protein have not yet been reported. In this study, a portable gel electrophoresis device for rapid separation and detection of proteins was developed and manufactured by 3D printing in a laboratory, which was economical, convenient, and quick. First, four kinds of portable gel electrophoresis devices that included three kinds of columnar gel and one slab gel electrophoresis device were designed with computer-aided design software SolidWorks 2017 (Dassault Systemes SE, France); the components including gel tubes, gel plates, and gel electrophoresis tanks were then printed using a 3D printer after optimization of the printing parameters. Then, the performance of the four kinds of gel electrophoresis devices was investigated using prestained protein molecular weight standards. The results showed that the single-channel slab gel electrophoresis design can quickly separate proteins with the best separation efficiency. Moreover, the effect of different separation gel lengths (5, 10, 15, and 20 mm) on protein separation was studied and it was found that 10% separation gels with a length of 5 mm could effectively separate prestained protein molecular weight standards (in the range of 15-250 kD) in 20 minutes. Next, the battery was optimized for the portable GE device and a 25 V lithium battery (70 mm×60 mm×40 mm) was used as the power supply, which could provide a constant voltage of 25 V for 100 hours during gel electrophoresis. Then, the One-Step Blue^TM reagent (Biotium, USA) was used to color the separation results of the five standard proteins (carbonic anhydrase, ovalbumin, bovine serum albumin, conalbumin, ribonuclease A), and the results were recorded by mobile phone. Finally, the proposed gel electrophoresis device was compared with the commercial device. The results showed that the two devices are comparable; however, the slab gel electrophoresis was faster, portable, and economical.
In summary, this research designed and manufactured a portable gel electrophoresis device using 3D printing technique, which can be used for on-site analysis and detection of proteins. The device presents the following advantages compared with the commercial devices:1) small and portable:the size of the electrophoresis tank of the device is only 15 mm×20 mm×17 mm and the 25 V lithium battery has a working time of approximately 100 hours; 2) low cost:it can be processed in 5 hours using 3D printing technology, with 10 mL of printing material while the total cost is less than 400 RMB; 3) fast separation:this device can quickly achieve protein separation compared with commercial devices and can further use multiple electrophoresis tanks in parallel to analyze more samples at the same time. Besides, this research also highlights the advantages of 3D printing for the development of miniaturized analytical equipment. Though this study has achieved preliminary results for rapid separation of proteins using gel electrophoresis devices, the quantitative analysis of proteins following protein detection and the application of more samples need further research. Meanwhile, the continued application of 3D printing technology will promote the development of miniaturized and portable experimental equipment.

Key words:

1. [1] Pan J Z, Fang P, Fang X X, et al. Sci Rep, 2018, 8(1):1791
2. [2] Guan Y F, Wang J W, Duan C F. Chinese Journal of Chromatography, 2009, 27(5):592 关亚风, 王建伟, 段春凤. 色谱, 2009, 27(5):592
3. [3] Badr I H A, Johnson R D, Madou M J, et al. Anal Chem, 2002, 74(21):5569
4. [4] Figeys D, Pinto D. Anal Chem, 2000, 72(9):330A
5. [5] Nachamkin I, Panaro N J, Li M, et al. J Clin Microbiol, 2001, 39(2):754
6. [6] Renzi R F, Stamps J, Horn B A, et al. Anal Chem, 2005, 77(2):435
7. [7] Skelley A M, Scherer J R, Aubrey A D, et al. Proc Natl Acad Sci U S A, 2005, 102(4):1041
8. [8] Liu R H, Lodes M J, Nguyen T, et al. Anal Chem, 2006, 78(12):4184
9. [9] Sista R, Hua Z, Thwar P, et al. Lab Chip, 2008, 8(12):2091
10. [10] Yuan Y X, Fan C, Pan J Z, et al. Chinese Journal of Chromatography, 2020, 38(2):183 袁颖欣, 樊晨, 潘建章, 等. 色谱, 2020, 38(2):183
11. [11] Lee B S, Lee Y U, Kim H S, et al. Lab Chip, 2011, 11(1):70
12. [12] Yin H F, Killeen K. J Sep Sci, 2007, 30(10):1427
13. [13] Li Y, Dvorak M, Nesterenko P N, et al. Anal Chim Acta, 2015, 896:166
14. [14] Cong W T, Zhou A, Liu Z G, et al. Anal Chem, 2015, 87(3):1462
15. [15] Hu L, Cheng T, He B, et al. Angew Chem Int Ed Engl, 2013, 52(18):4916
16. [16] Yan X T, He B, Wang D Y, et al. Talanta, 2018, 184:404
17. [17] Zhu G L, Yang Z H. Plant Physiology Communications, 1982(2):43 朱广廉, 杨中汉. 植物生理学通讯, 1982(2):43
18. [18] Shi J H, Zhao Y T, Wang J L, et al. Journal of the Fourth Military Medical University, 2000, 21(6):761 石继红, 赵永同, 王俊楼, 等. 第四军医大学学报, 2000, 21(6):761
19. [19] Zhao X, Yang L, Yu Z, et al. J Environ Sci, 2008, 20(2):224
20. [20] Wang D Y, He B, Yan X T, et al. Talanta, 2019, 197:145
21. [21] Gross B, Lockwood S Y, Spence D M. Anal Chem, 2017, 89(1):57
22. [22] Dixit C K, Kadimisetty K, Rusling J. TrAC-Trends Anal Chem, 2018, 106:37
23. [23] Gaal G, Mendes M, De Almeida T P, et al. Sens Actuator B-Chem, 2017, 242:35
24. [24] Su C K, Hsieh M H, Sun Y C. Anal Chim Acta, 2016, 914:110
25. [25] Lücking T H, Sambale F, Beutel S, et al. Eng Life Sci, 2015, 15(1):51
26. [26] Simon U, Dimartino S. J Chromatogr A, 2019, 1587:119
27. [27] Calderilla C, Maya F, Cerda V, et al. Talanta, 2019, 202:67
28. [28] Macdonald N P, Currivan S A, Tedone L, et al. Anal Chem, 2017, 89(4):2457
29. [29] Zhang C, Glaros T, Manicke N E. J Am Chem Soc, 2017, 139(32):10996
30. [30] Dou M, Sanchez J, Tavakoli H, et al. Anal Chim Acta, 2019, 1065:71
31. [31] Rong Z, Wang Q, Sun N X, et al. Anal Chim Acta, 2019, 1055:140
32. [32] Hu L G, Jiang G B. Environ Sci Technol, 2017, 51(7):3597
33. [33] Wang D, Huang X, Li J, et al. Chem Commun, 2018, 54(22):2723
34. [34] Au A K, Huynh W, Horowitz L F, et al. Angew Chem Int Edit, 2016, 55(12):3862
35. [35] Ho C M B, Ng S H, Li K H H, et al. Lab Chip, 2015, 15(18):3627
36. [36] Yazdi A A, Popma A, Wong W, et al. Microfluid Nanofluid, 2016, 20(3):50
37. [37] Zhang L, Wang W, Ju X J, et al. RSC Adv, 2015, 5(8):5638
38. [38] Lupton D. Soc Theory Health, 2015, 13(2):99

扫一扫看文章

计量

PDF下载量: 2
文章访问数: 85
HTML全文浏览量: 8

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

3D打印便携式凝胶电泳装置用于蛋白质的快速检测

通讯作者: 胡立刚, E-mail:lghu@rcees.ac.cn

English

3D printed portable gel electrophoresis device for rapid detection of proteins

Corresponding author: HU Ligang, lghu@rcees.ac.cn

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

3D打印便携式凝胶电泳装置用于蛋白质的快速检测

通讯作者: 胡立刚, E-mail:lghu@rcees.ac.cn

English

3D printed portable gel electrophoresis device for rapid detection of proteins

Corresponding author: HU Ligang, lghu@rcees.ac.cn

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs