Journal
ELECTROPHORESIS
Volume 38, Issue 13-14, Pages 1706-1712Publisher
WILEY
DOI: 10.1002/elps.201600472
Keywords
Computer simulation; Flow venting; Moving reaction boundary electrophoresis; Protein titration
Funding
- National Natural Science Foundation of China [21275099, 21305087, 21475086]
- National Key Development of Scientific Instruments [2011YQ030139]
- Shanghai Science and Technology Committee [14DZ0501200, 15142200300]
- Shanghai Jiao Tong University [YG2010ZD209, YG2014QN21]
- Recruitment Program of Global Youth Experts of China
- National High-tech R&D Program of China (863 Program) [2014AA020545]
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Moving reaction boundary titration (MRBT) has a potential application to immunoassay and protein content analysis with high selectivity. However, air bubbles often impair the accuracy of MRBT, and the leakage of electrolyte greatly decreases the safety and convenience of electrophoretic titration. Addressing these two issues a reliable MRBT device with modified electrolyte chamber of protein titration was designed. Multiphysics computer simulation was conducted for optimization according to two-phase flow. The single chamber was made of two perpendicular cylinders with different diameters. After placing electrophoretic tube, the resident air in the junction next to the gel could be eliminated by a simple fast electrolyte flow. Removing the electrophoretic tube automatically prevented electrolyte leakage at the junction due to the gravity-induced negative pressure within the chamber. Moreover, the numerical simulation and experiments showed that the improved MRBT device has following advantages: (i) easy and rapid setup of electrophoretic tube within 20 s; (ii) simple and quick bubble dissipates from the chamber of titration within 2 s; (iii) no electrolyte leakage from the two chambers: and (iv) accurate protein titration and safe instrumental operation. The developed technique and apparatus greatly improves the performance of the previous MRBT device, and providing a new route toward practical application.
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