Multichannel flow electrophoresis and its applications in purification of proteins, enzymes, and antibodies.
Multichannel flow electrophoresis (MFE) is a novel preparative electrophoresis technique designed for continuous separation of proteins based on their different isoelectric points. A computer controlled MFE apparatus is developed and the effects of operation parameters such as electric field strength, buffering pH, and sample introducing flow rate on MFE separation efficiency are investigated. The application of MFE is first exemplified by continuous separation of BSA and HBB mixture. Then continuous purification of urokinase from urine extracts is conducted, in which 10 x 10(4) IU urokinase product whose specific activity is higher than 40,000 IU/mg is obtained per hour. Finally, MFE is applied in the continuous purification of anti-urokinase mouse IgG from mouse serum and yields 2.1 mg mouse IgG per hour with high purity demonstrated by SDS-PAGE. The preliminary results presented have confirmed the workability of MFE and pointed out its high application potential in biochemical process.
Liu
Z
Z
Department of Chemical Engineering, Tsinghua University, Beijing, China.
Feng
S
S
Ding
F
F
Yuan
N
N
D000906:Antibodies / Q000302:isolation & purification*; D004586:Electrophoresis / Q000379:methods*; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification*; D014568:Urokinase-Type Plasminogen Activator / Q000276:immunology / Q000302:isolation & purification* / Q000652:urine
D000906:Antibodies; D012710:Serum Albumin, Bovine; D014568:Urokinase-Type Plasminogen Activator