Application of avidin-biotin technology for the characterization of a model hapten-protein conjugate.
A simple method was developed for the rapid characterization of the covalent binding of haptens to proteins such as enzymes, bovine serum albumin (BSA), and other carrier-proteins and antibodies. In the present study, a commercially available fentanyl-BSA conjugate was characterized by a 4'-hydroxyazobenzene-2-carboxylic acid (HABA) dye assay that followed a biotinylation reaction. This protocol allowed the indirect observation of the average hapten number per BSA molecule. Such measurement is useful for optimizing reaction conditions to yield a more precisely defined product for immunological applications. The obtained result was within the limits suggested by the manufacturer of the conjugate.
Dotsikas
Yannis
Y
Department of Pharmaceutical Chemistry, School of Pharmacy, University of Athens, Panepistimioupoli Zografou, GR-157 71, Athens, Greece.
Loukas
Yannis L
YL
D001360:Avidin / Q000737:chemistry*; D001391:Azo Compounds; D019921:Biotinylation / Q000379:methods*; D005283:Fentanyl / Q000031:analogs & derivatives* / Q000276:immunology; D005456:Fluorescent Dyes; D006241:Haptens / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000276:immunology; D021241:Spectrometry, Mass, Electrospray Ionization
D001391:Azo Compounds; D005456:Fluorescent Dyes; D006241:Haptens; D011506:Proteins; C505444:fentanyl-BSA conjugate; D001360:Avidin; C001413:HABA; D012710:Serum Albumin, Bovine; D005283:Fentanyl