Parallel immunizations of rabbits using the same antigen yield antibodies with similar, but not identical, epitopes.

ABSTRACT: A problem for the generation of polyclonal antibodies is the potential difficulties for obtaining a renewable resource due to batch-to-batch variations when the same antigen is immunized into several separate animals. Here, we have investigated this issue by determining the epitopes of antibodies generated from parallel immunizations of rabbits with recombinant antigens corresponding to ten human protein targets. The epitopes were mapped by both a suspension bead array approach using overlapping synthetic 15-mer peptides and a bacterial display approach using expression of random fragments of the antigen on the surface of bacteria. Both methods determined antibody binding with the aid of fluorescent-based analysis. In addition, one polyclonal antibody was fractionated by peptide-specific a

SUBMITTER: Hjelm B 

PROVIDER: S-EPMC3526615 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature