Conjucation of R-Phycoerythrin to a polyclonal antibody and F [ab'] 2 fragment of a polyclonal antibody by two different methods

ABSTRACT

R-phycoerythrin [R-PE], a fluorescent protein from phycobiliprotein family, is isolated from red algae. Conjugation of antibodies to R-PE facilitates multiple fluorescent staining methods. In the present study polyclonal antibodies and polyclonal F[ab']2 fragment antibodies were conjugated to R-PE by two different methods. The efficiency of the methods was evaluated using Immunocytochemistry [ICC] and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis [SDS-PAGE]. In the first conjugation method, PE was attached to SMCC linker followed by conjugation of antibody to PE-SMCC. In the second method, SH groups were added onto R-PE molecule, while the antibody was attached to SPDP linker. Then, the antibody-SPDP molecule was conjugated to R-PE. Our results showed that the two conjugation methods did not have any abrogative effects on the antibody binding activity