Use of soybean peroxidase in chemiluminescent enzyme-linked immunosorbent assay.
A procedure for the production of conjugates of soybean peroxidase (SbP) oxidized by sodium periodate and anti-mouse IgG antibody (Ab) was optimized. A sandwich chemiluminescent enzyme-linked immunosorbent assay (ELISA) for determination of mouse IgG using SbP and specific Ab was developed, and SbP-catalyzed oxidation of luminol was carried out in the absence of any enhancer. Comparison of conjugates produced by labeling Ab by soybean and horseradish peroxidases in the chemiluminescent ELISA showed that in the case of SbP a rate of emission decay formed through luminol oxidation was significantly lower. Application of the soya enzyme allowed the development of the immunoassay with improved sensitivity and a wider linear range.
Sakharov
Ivan Yu
IY
Chemical Enzymology Department, Faculty of Chemistry, The M.V. Lomonosov Moscow State University, Russia. sakharov@enz.chem.msu.ru
Alpeeva
Inna S
IS
Efremov
Evgeny E
EE
D000818:Animals; D000888:Antibodies, Anti-Idiotypic; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006735:Horseradish Peroxidase; D007074:Immunoglobulin G / Q000032:analysis / Q000276:immunology; D008163:Luminescent Measurements; D051379:Mice; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000737:chemistry; D009195:Peroxidase; D012680:Sensitivity and Specificity; D013025:Glycine max / Q000201:enzymology*
D000888:Antibodies, Anti-Idiotypic; D007074:Immunoglobulin G; D010504:Periodic Acid; C009288:metaperiodate; D006735:Horseradish Peroxidase; D009195:Peroxidase