A simple and highly stable free-flow electrophoresis device with thermoelectric cooling system.
Complex assembly, inconvenient operations, poor control of Joule heating and leakage of solution are still fundamental issues greatly hindering application of free-flow electrophoresis (FFE) for preparative purpose in bio-separation. To address these issues, a novel FFE device was developed based on our previous work. Firstly, a new mechanical structure was designed for compact assembly of separation chamber, fast removal of air bubble, and good anti-leakage performance. Secondly, a highly efficient thermoelectric cooling system was used for dispersing Joule heating for the first time. The systemic experiments revealed the three merits: (i) 3min assembly without any liquid leakage, 80 times faster than pervious FFE device designed by us or commercial device (4h); (ii) 5s removing of air bubble in chamber, 1000-fold faster than a normal one (2h or more) and (iii) good control of Joule heating by the cooling system. These merits endowed the device high stable thermo- and hydro-dynamic flow for long-term separation even under high electric field of 63V/cm. Finally, the developed device was used for up to 8h continuous separation of 5mg/mL fuchsin acid and purification of three model proteins of phycocyanin, myoglobin and cytochrome C, demonstrating the applicability of FFE. The developed FFE device has evident significance to the studies on stem cell, cell or organelle proteomics, and protein complex as well as micro- or nano-particles.
School of Life Science and Biotechnology, Shanghai 200240, China; Institute of Refrigeration and Cryogenics, School of Mechanical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China.
Jian
J
Yan
Cheng-Gang
CG
Guo
Xiao-Ping
XP
Liu
Fan-Zhi
FZ
Kong
Qiao-Yi
QY
Shen
Cheng-Zhang
CZ
Yang
Jun
J
Li
Cheng-Xi
CX
Cao
Xin-Qiao
XQ
Jin
D003080:Cold Temperature; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation*; D004867:Equipment Design
Free-flow electrophoresis; Joule heating; Separation chamber; Thermoelectric cooler