Protein separation with aqueous-aqueous polymer systems by two types of counter-current chromatographs.
Two different types of counter-current chromatographs, the cross-axis coil planet centrifuge (X-axis CPC) and horizontal flow-through coil planet centrifuge (horizontal CPC), were evaluated for protein separation with an aqueous-aqueous two-phase polymer system. The sample solution, containing 10-200 mg each of cytochrome c, myoglobin, ovalbumin and hemoglobin in 2 ml of each phase was eluted with the lower phase. In both instruments, the effects of the flow-rate, revolution speed, and parameter beta (helical diameters of the multilayer coil) on the protein separation were investigated. The best results were obtained from the X-axis CPC operated at 750 rpm and a flow-rate of 2.0 ml/min using a multilayer coil with a small helical diameter (beta = 0.25-0.60). Four protein samples were well resolved in less than 5 h.
Shibusawa
Y
Y
Laboratory of Biophysical Chemistry, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892.
Ito
Y
Y
D000818:Animals; D002417:Cattle; D002645:Chickens; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D003377:Countercurrent Distribution; D003574:Cytochrome c Group / Q000032:analysis; D006454:Hemoglobins / Q000032:analysis; D006736:Horses; D006863:Hydrogen-Ion Concentration; D009211:Myoglobin / Q000032:analysis; D010047:Ovalbumin / Q000032:analysis; D011108:Polymers / Q000032:analysis*; D011506:Proteins / Q000032:analysis*
D003574:Cytochrome c Group; D006454:Hemoglobins; D009211:Myoglobin; D011108:Polymers; D011506:Proteins; D010047:Ovalbumin