Electrochemical reduction of disulfide bonds in proteinaceous substances and electrochemical cell for carrying out such reduction

摘要

The invention provides a method of cleaving disulfide bonds in proteinaceous substances by means of electrochemical reduction, said method comprising: €¢ providing a liquid sample containing one or more proteinaceous substances that comprises at least one disulfide bond; €¢ providing an electrochemical cell comprising a working electrode and an auxiliary electrode; €¢ introducing the liquid sample into the electrochemical cell; €¢ subjecting the liquid sample in the electrochemical cell to a reducing potential to reduce disulfide bonds in the one or more proteinaceous substances, thereby producing an electrochemically reduced liquid sample containing proteinaceous substances with a cleaved disulfide bond; and €¢ further processing the electrochemically reduced liquid sample; wherein the electrochemical cell comprises a working electrode that contains titanium. The invention further relates to an electrochemical flow cell for processing a sample fluid, the cell comprising: €¢ a body having a flow path, the flow path having an inlet and an outlet; €¢ a working electrode in fluid communication with the flow path; and €¢ an auxiliary electrode; wherein the working electrode comprises a component or layer that contains at least 20 wt.% titanium in the form of elemental titanium, titanium-containing substances and/or titanium-containing alloys; and wherein the outlet of the cell is connected to an electrochemical detection (ECD) device, a NMR spectrometer or a mass spectrometer (MS) in case the working electrode is a ruthenium-plated titanium electrode. The use of a working electrode containing titanium enables fast, stable and substantially complete reduction of all disulfide bonds in proteinaceous substances.