title: Electrochemistry and chemiluminescence techniques compared in the detection of NADPH oxidase activity in phagocyte cells.
abstract: Several methodologies have been used in clinical chemistry for real-time assessment of NADPH oxidase primary product superoxide anion which dismutases to hydrogen peroxide. Among these methodologies, isoluminol chemiluminescence (CL) is considered to be one of the more sensitive and reliable techniques for the assessment of NADPH oxidase activity in neutrophils. The electrochemical technique was recently designed and also applied for real-time detection of NADPH oxidase activity in neutrophils but its reliability and sensitivity has not been investigated so far. In this study, isoluminol CL and electrochemical techniques were investigated and compared by monitoring the generation of superoxide and hydrogen peroxide in both PLB 985 cell line differentiated into neutrophil-like cells and human neutrophils. The electrochemical technique was shown to be as sensitive as that of CL and able to detect the reactive oxygen species (ROS) release of as low as 500 cells. Thus, the electrochemical technique could be used as an alternative to optical techniques for the evaluation of extracellular ROS in phagocyte cells.
authors.affiliation: Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva, Israel. ashkenaa@bgu.ac.il
authors.forename: A
authors.initials: A
authors.lastname: Ashkenazi
authors.forename: K
authors.initials: K
authors.lastname: Abu-Rabeah
authors.forename: R S
authors.initials: RS
authors.lastname: Marks
mesh_terms: D004563:Electrochemistry / Q000379:methods*; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis; D007700:Kinetics; D049449:Luminescence; D008163:Luminescent Measurements / Q000379:methods*; D008956:Models, Chemical; D019255:NADPH Oxidases / Q000032:analysis*; D009504:Neutrophils / Q000378:metabolism; D010586:Phagocytes / Q000378:metabolism*; D017382:Reactive Oxygen Species; D012680:Sensitivity and Specificity; D013481:Superoxides / Q000378:metabolism; D013997:Time Factors
chemical_list: D017382:Reactive Oxygen Species; D013481:Superoxides; D006861:Hydrogen Peroxide; D019255:NADPH Oxidases