Colorimetric competitive inhibition method for the quantification of avidin, streptavidin and biotin.
A colorimetric competitive inhibition assay for avidin, streptavidin and biotin was developed. The method for avidin or streptavidin was based on the competitive binding between avidin or streptavidin and a streptavidin-enzyme conjugate for biotinylated dextrin immobilized on the surface of a microtitre plate. For biotin quantitation the competition is between free biotin and the immobilized biotin for the streptavidin-enzyme conjugate. The limits of detection which was determined as the concentration of competitor required to give 90% of maximal absorbency (100% inhibition) was approximately 20 ng/100 microl per assay for avidin and streptavidin and 0.4 pg/100 microl per assay for biotin. The methods are simple, rapid, highly sensitive and adaptable to high throughput analysis.
Gan
Z
Z
Department of Animal Science, The University of Manitoba, Winnipeg, Canada.
Marquardt
R R
RR
D001360:Avidin / Q000032:analysis* / Q000037:antagonists & inhibitors*; D001667:Binding, Competitive; D001710:Biotin / Q000032:analysis* / Q000037:antagonists & inhibitors*; D003124:Colorimetry / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D019809:Streptavidin / Q000032:analysis* / Q000037:antagonists & inhibitors*
D001360:Avidin; D001710:Biotin; D019809:Streptavidin