Influence of Antibody affinity on the performance of different antibody assays
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2010, VaccineCitation Excerpt :At the end of each dilution, the bound conjugate was visualized with 3,3,5,5′-tetramethylbenzidine (TMB) substrate which gave a blue reaction product, the intensity of which was proportional to the concentration of antibodies in the sample and this was read by an optical instrument (Bio Rad M500, Bio Rad, France). Even though in principle an ELISA can be used for the determination of antibody affinity, the ELISA test kit which we used was a basic one which measured antibody concentration at a given dilution [2,9]. Different dilutions of antibody concentrations were used to plot graphs against the corresponding optical densities to derive dose–response curves which were then used to determine midpoint titres which could serve as a proxy for affinity.
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