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Antec introduced the ROXY as an electrochemical reactor up-front MS. The electrochemically-assisted reduction of disulphide bonds followed by on-line mass spectrometric (MS) detection is presented.
Antec introduced the ROXY as an electrochemical reactor up-front MS. The electrochemically-assisted reduction of disulphide bonds followed by on-line mass spectrometric (MS) detection is presented. The method is based on square-wave potential pulses applied on a new type of proprietary working electrode made from titanium alloy. The method does not use any chemical agents and is purely instrumental, resulting in a fully automated platform for fast assessment and characterization of S–S bonds in proteins and peptides.
Figure 1: A schematic of a disulphide bond reduction: Replacing interfering chemicals (for example, DTT, TCEP) by an electrochemical reactor cell used on-line with LC–MS.
An electrochemical reactor offers an advantage over chemical reactions. By changing the applied potential, the extent of disulphide bond cleavage is controlled. More negative potentials result in a shift of the charge state distribution, thereby indicating increased disulphide bond cleavage and unfolding of the protein.
Figure 2: Partial and full reduction of disulphide bonds in α-lactalbumin. The overlapping isotopic pattern of the +9 ion measured with the EC reactor cell turned OFF (top), and ON at E1 = -1000 mV and E1 = -1300 mV.
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