Optimized continuous flow electrophoresis

Continuous flow electrophoresis (CFE) was optimized by employing (i) electrophoretic regimes with stacking properties, to eliminate electrohydrodynamic dispersion, (ii) quasi‐mixed zones to prevent precipitation of the stacked analytes, (iii) sheath liquid streams at the electrode compartment membranes to prevent penetration of the electrode reaction products into the separation chamber, (iv) proper engineering of the separation chamber to provide efficient dissipation of Joule heat, and (v) counterflow at the collection outlets to eliminate the problems of dead volumes and uneven collection of separated species. Data on direct temperature measurements in the separation chamber at various levels of the dissipated electric power are presented. Preparative runs of amyloglucosidase in the isoelectric focusing (IEF) mode and rat liver organelles in the isotochophoresis (ITP) mode demonstrate the high performance of the optimized CFE system.