Separation of metallothionein isoforms with capillary zone electrophoresis using an uncoated capillary column : Effects of pH, temperature, voltage, buffer concentration and buffer composition
Metallothionein (MT) isoforms, sulphydryl-rich proteins with a high affinity for both essential and toxic trace metals, are readily separated by capillary zone electrophoresis (CZE) with an uncoated polyimine-clad fused-silica capillary using Tris-borate buffer. The metallothionein samples investigated were rabbit liver MT, rabbit liver MT-1, rabbit liver MT-2 and horse kidney MT. The effects of temperature, buffer pH, buffer concentration, buffer composition and running voltage on the separation efficiency were investigated. An improved separation efficiency compared with published methods using uncoated fused-silica capillaries was obtained, allowing the separation of peaks which previously co-migrated. The running conditions used were voltage 11 kV, temperature 20 C and buffer 110 mM Tris-110 mM borate (pH 6.90). Under these conditions, horse kidney MT exhibited five putative isoform peaks and rabbit liver MT nine or ten peaks. Interestingly, the experiments indicated that rabbit liver MT contains one major component which is not found in either isoform MT-1 or MT-2, possibly indicating that the improved separation efficiency made it possible to detect a new group or structure. Based on this initial evaluation. CZE with an uncoated fusion-silica capillary using Tris-borate buffer appears to be a very useful method for the separation of metallothionein isoforms.
Bibliographic Reference: Article: Journal of Chromatography A Vol. 734 (1996), pp. 391-400
Record Number: 199611018 / Last updated on: 1996-09-30
Original language: en
Available languages: en