Development and characterization of monoclonal antibodies to chicken riboflavin carrier protein

Several monoclonal antibodies (MAbs) specific to chicken riboflavin carrier protein (cRCP) were developed and characterized. Of the several MAbs analyzed, four were directed against nonoverlapping epitopes as demonstrated by MAb inhibition assay. Many of these epitopes appeared to be in close proximity and only three were situated at distinct part of the molecule as revealed by sandwich assay. A combination of chemical modification, peptide cleavage by chemical and enzymatic methods, was used to analyze the possible antigenic structure recognized by these MAbs. An assembled epitope spanning the region 22-87 forms the antigenic site recognized by 4999.1; while MAb 5555.3 interacted with the C-terminal peptide 203-219.