Detection system for membrane immunoassay based on the trapping of a highly colored intermediate of the peroxidase reaction.
The oxidation of ortho-dianisidine by membrane bound horseradish peroxidase in the presence of sodium dextran sulfate affords a dark-green insoluble product, identified as an unstable meriquinone intermediate previously reported in literature. Cationic and unsubstituted dextrans do not stabilize the intermediate. The highest yield of the intermediate is observed at pH 4.0-5.0 and concentration of dextran sulfate ca. 0.5%. New highly sensitive detection system for peroxidase has been developed on the basis of ortho-dianisidine oxidation to the meriquinone intermediate in the presence of sodium dextran sulfate. Under certain conditions, with lowered sodium dextran sulfate concentrations, a progressive further oxidation of the green intermediate to the yellow-brown final product is observed on passing to higher enzyme concentrations. This finding opens a possibility to develop a detection system in which the color of the mixture of reaction products serves as a measure of the enzyme concentration.
Yatsimirskaya
E A
EA
Department of Chemistry, Moscow State University, Leninskye Gory, Russia.
Gavrilova
E M
EM
Egorov
A M
AM
D002623:Chemistry Techniques, Analytical / Q000379:methods*; D016264:Dextran Sulfate; D003962:Dianisidine / Q000032:analysis*; D006735:Horseradish Peroxidase / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques; D008567:Membranes, Artificial; D009757:Nylons; D010084:Oxidation-Reduction; D013053:Spectrophotometry / Q000379:methods
D008567:Membranes, Artificial; D009757:Nylons; D016264:Dextran Sulfate; D006735:Horseradish Peroxidase; D003962:Dianisidine