Biochemiluminescence sensing of protein oxidation by reactive oxygen species generated by pulsed electric field

Pulsed electric field (PEF) technology is an important emerging modality for both biomedicine and the food industry, yet the mechanisms of action of a PEF on a biomolecular level remain unclear. In order to gain a better understanding of this, a sensing platform was developed to enable sensitive detection of oxidative processes induced by the PEF. It was found that the application of PEF in phosphate buffer leads to the electrogeneration of reactive oxygen species (ROS) which in turn triggers intrinsic chemiluminescence. Furthermore, the presence of the bovine serum albumin (BSA) was found to increase the luminescence signal, which was attributed to biochemiluminescence due to the reactions of BSA with the electrogenerated ROS. A carbonyl determination assay was then used to confirm that the BSA molecules had indeed been oxidatively modified. We proposed a scheme describing the reactions leading from charge transfer at the anode/electrolyte interface to the actual photon emission, helping to elucidate the mechanisms of action of PEF on proteins. The results may to open up new avenues for the application of PEF technology.