Highly sensitive chemiluminescence method for determining myeloperoxidase in human polymorphonuclear leukocytes.
Myeloperoxidase activity was assayed by a chemiluminescence method, using a cypridina luciferin analog as a chemiluminescence probe, after extraction from peripheral human polymorphonuclear leukocytes. The chemiluminescence method was based on the detection of 1O2 generated by myeloperoxidase-catalyzed HOBr formation followed by the interaction of HOBr with H2O2 at pH 4.5. With this method, myeloperoxidase in less than 100 polymorphonuclear leukocytes could be detected and myeloperoxidase in 10(6) polymorphonuclear leukocytes would be calculated to be 14.4 pmol. Eosinophil extract, which contains eosinophil peroxidase, catalyzed 1O2 generation to a great extent, compared with the polymorphonuclear leukocyte extract at pH 4.5. Myeloperoxidase activity in extract of neutrophil fraction could be greatly influenced by eosinophil contamination.
Uehara
K
K
Photon Medical Research Center, Hamamatsu University School of Medicine, Shizuoka, Japan.
Hori
K
K
Nakano
M
M
Koga
S
S
D003676:Deferoxamine / Q000378:metabolism; D005609:Free Radicals; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007093:Imidazoles / Q000378:metabolism; D009504:Neutrophils / Q000201:enzymology*; D010100:Oxygen / Q000378:metabolism; D009195:Peroxidase / Q000032:analysis*; D011719:Pyrazines / Q000378:metabolism; D012680:Sensitivity and Specificity; D013347:Subcellular Fractions; D013379:Substrate Specificity; D013482:Superoxide Dismutase / Q000378:metabolism
D005609:Free Radicals; D007093:Imidazoles; D011719:Pyrazines; C063412:2-methyl-6-(4-methoxyphenyl)-3,7-dihydroimidazo(1,2-alpha)pyrazin-3-one; D006861:Hydrogen Peroxide; D009195:Peroxidase; D013482:Superoxide Dismutase; D003676:Deferoxamine; D010100:Oxygen