A COMPARATIVE-STUDY OF PEROXIDASES FROM HORSE RADISH AND ARTHROMYCES-RAMOSUS AS LABELS IN LUMINOL-MEDIATED CHEMILUMINESCENT ASSAYS

被引:55
作者
KIM, BB
PISAREV, VV
EGOROV, AM
机构
[1] Division of Chemical Enzymology, Chemistry Department, Lomonosov Moscow University
关键词
D O I
10.1016/0003-2697(91)90260-Z
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The properties of a peroxidase from Arthromyces ramosus (ARP) in the chemiluminescent reaction of luminol oxidation have been studied. These were compared with the properties of horse radish peroxidase (HRP) in the cooxidation of luminol and p-iodophenol, the enhanced chemiluminescence (ECL) reaction. By means of the stop-flow technique, ARP was shown to have an enzymatic activity toward luminol higher than that toward HRP. ARP can efficiently catalyze luminol oxidation in the absence of substrate enhancer. pH and substrate concentrations were optimized to determine ARP with the highest sensitivity. The detection limit of ARP was 5 × 10-13 m, the same as that for HRP in the ECL reaction. The data on the use of ARP as a label in enzyme immunoassay of human IgG are presented. ARP was shown to have all the advantages of HRP as a label in chemiluminescent enzyme immunoassays: (i) high signal intensity, (ii) slow decay of luminescence, (iii) high signal/noise ratio, and (iv) as a consequence of (i)-(iii), high detection sensitivity. However, the low thermostability of ARP can limit the potential fields of its application. © 1991.
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页码:1 / 6
页数:6
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