Preparative isoelectric focusing and Joule effect: a purification cell that contains a heat exchanger.
Reproducibility in protein purification by preparative isoelectric focusing depends greatly on temperature control during the separation process. A preparative apparatus is described, including a heat exchanger between compartments with isoelectric membranes. The selectivity of the isoelectric membranes was optimized as a function of isoelectric points of the separated proteins. At 2500 V and 60 W, 0.3 g of horse heart myoglobin from 0.2 g of whale skeletal muscle myoglobin could be separated in 1 h. At a total load of 2 g protein, 97% of bovine hemoglobin (2% initial concentration) was purified from bovine serum albumin (0.15%).
Institute of Chemical Engineering, Swiss Federal Institute of Technology, Lausanne, Switzerland.
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Araque
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Jaugey
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Javet
D004563:Electrochemistry; D004867:Equipment Design; D007525:Isoelectric Focusing / Q000295:instrumentation*; D011506:Proteins / Q000302:isolation & purification*; D015203:Reproducibility of Results; D013696:Temperature; D013816:Thermodynamics
D011506:Proteins