Novel fluorometric protocol for assessing myeloperoxidase activity.

Autor: Majid R; Department of Medical Physics, University of Al-Mustaqbal, Hilla City, Babylon Governorate p.o. 51001, Iraq., Al Talebi ZA; Chemistry Dept., College of Science, University of Babylon, Hilla City, Babylon Governorate p.o. 51002, Iraq., Al-Kawaz HS; Department of Pathological Analysis, College of Science, Al-Qasim Green University, 51013, Iraq., Hassan Alta'ee A; College of Medicine, University of Babylon, Hilla City, Babylon Governorate, Iraq., Alsalman ARS; College of Medicine, University of Babylon, Hilla City, Babylon Governorate, Iraq., Hadwan AM; Faculty of Medicine, Golestan University of Medical Sciences, Gorgan, Iran; Al-Manara College for Medical Sciences, Al-Amarah City, Iraq., Hadwan MM; College of Medicine, University of Babylon, Hilla City, Babylon Governorate, Iraq., Hadwan MH; Chemistry Dept., College of Science, University of Babylon, Hilla City, Babylon Governorate p.o. 51002, Iraq. Electronic address: mahmoudhadwan@gmail.com.
Jazyk: angličtina
Zdroj: Enzyme and microbial technology [Enzyme Microb Technol] 2023 Dec; Vol. 171, pp. 110320. Date of Electronic Publication: 2023 Sep 09.
DOI: 10.1016/j.enzmictec.2023.110320
Abstrakt: Neutrophil myeloperoxidase (MPO) is an essential enzyme for the innate immune system. Measuring MPO activity is vital for understanding neutrophil characteristics and functions in various diseases. MPO activity can be measured using several methods, including spectrophotometric and fluorometric protocols. This paper introduces a fluorometric method for specifically quantifying MPO activity based on the H 2 O 2 -dependent oxidation of thiamine. We optimized this new method using the robust statistical approach response surface methodology (RSM) and Box Benken Design (BBD). We extensively examined the effects of several experimental parameters using the RSM methodology and determined the best conditions for accurate and sensitive MPO activity measurement. The optimal conditions were determined using the analysis of variance (ANOVA) for second-order polynomial equations. The resulting F-value (4.86) indicated that the model was significant. However, the lack-of-fitness F-value (1.79) suggested it did not differ significantly from the corresponding p-value. The greatest MPO activity (30 ± 2 U L -1 ) was obtained under optimum conditions, which were 1000 µM of H 2 O 2 , 10 min incubation time, and 1000 µM of thiamine. Our results suggest that this advanced fluorometric method has significant accuracy, sensitivity, and linearity up to 60 IU. The new and standard colorimetric methods also showed a good correlation. These results indicate that the new fluorometric method can be dependable and efficient for assessing MPO activity. The new method is characterized by excellent accuracy, sensitivity, and linearity, making it a valuable protocol for researchers and clinicians interested in assessing MPO activity.
Competing Interests: Conflict of interest The authors declare no conflicts of interest or competing interests.
(Copyright © 2023 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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