Improved presentation of antigenic sites in enzyme-linked immunosorbent assay: antigenicity of reduced and carboxymethylated riboflavin carrier protein.
Disulphide reduced and carboxymethylated riboflavin carrier protein (RCM-RCP), an unfolded derivative of chicken RCP, does not bind riboflavin and there is a drastic reduction in its ability to interact with antiserum to cRCP. Antibodies to RCM-RCP are directed against sequential epitopes(s). On radioiodination of RCM-RCP, a maximum of 30-50% binding at dilution of 1:500 was obtained with rabbit antibodies RCM-RCP [n = 5]. However, high titer antibodies was obtained when radioiodinated RCM-RCP was immobilized on ELISA microtiter plates suggesting that immobilization of RCM-RCP leads to either preservation of antigenic sites or improved presentation of the antigenic determinants. An avidin-biotin system was utilized to develop an ELISA.
Pereira
J
J
Institute for Research in Reproduction (ICMR), Parel, Bombay.
Brahmbhatt
N D
ND
Natraj
U
U
D000818:Animals; D000937:Antigen-Antibody Reactions; D002352:Carrier Proteins / Q000737:chemistry* / Q000276:immunology; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D000939:Epitopes / Q000032:analysis*; D005260:Female; D007106:Immune Sera; D026901:Membrane Transport Proteins; D008745:Methylation; D010084:Oxidation-Reduction; D012256:Riboflavin / Q000378:metabolism
D002352:Carrier Proteins; D000939:Epitopes; D007106:Immune Sera; D026901:Membrane Transport Proteins; C019079:riboflavin-binding protein; D012256:Riboflavin