Author/Authors :
Mahmoudian، Jafar نويسنده Department of Immunochemistry, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran , , Jeddi Tehrani]، Mahmood نويسنده , , Rabbani، Hodjattallah نويسنده Immune and Gene Therapy Laboratory, CCK, Karolinska University Hospital Solna Department of Antigen and Antibody Engineering, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR , , Mahmoudi، Ahmad Reza نويسنده Department of Immunochemistry, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran , , Akhondi، Mohammad-Mehdi نويسنده , , Zarnani، Amir Hassan نويسنده Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, IR Iran , , Balaei Goli، Leila نويسنده Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR , , Babaei، Mahdokht نويسنده Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR , , Ghods، Roya نويسنده Department of Immunochemistry, Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran ,
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(abʹ)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.
