4] Structural characterization of hemoglobin variants using capillary electrophoresis and fast atom bombardment mass spectrometry

Abstract

Publisher Summary The combination of capillary zone electrophoresis (CZE) with fast atom bombardment mass spectrometry (FAB/MS) analysis according to the FAB mapping procedure couples a potentially fast high-resolution separation method with one of the most powerful techniques for structural characterization. Capillary electrophoresis has the ability to handle very small volume samples (picoliter range) and attomole (10 -18 mol)-level detection limits reaching separation efficiencies of the order of 10 6 theoretical plates. The CZE of proteins with UV detection have often resulted in broad, tailing peaks because of the absorption of proteins on active sites of the negatively charged capillary wall. However, the introduction of coated capillaries made CZE a fast and sensitive method for the detection of even similarly charged variants; the resolution obtained using this technique is comparable to the well-established electrophoretic and chromatographic procedures and requires smaller samples. Its speed and simplicity with respect to the given techniques and the possibility of automation and of development on a micropreparative scale mean the whole procedure is a significant alternative to conventional systems.