Mahmoudian Jafar, Jeddi-Tehrani Mahmood, Rabbani Hodjattallah, Mahmoudi Ahmad Reza, Akhondi Mohammad Mehdi, Zarnani Amir Hassan, Goli Leila Balaei, Babaei Mahdokht, Ghods Roya
Monoclonal Antibody Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
Avicenna J Med Biotechnol. 2010 Apr;2(2):87-91.
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.
藻红蛋白（R-PE）是一种来自藻胆蛋白家族的荧光蛋白，从红藻中分离得到。将抗体与R-PE偶联有助于多种荧光染色方法。在本研究中，通过两种不同方法将多克隆抗体和多克隆F(ab')2片段抗体与R-PE偶联。使用免疫细胞化学（ICC）和十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）评估这些方法的效率。在第一种偶联方法中，PE连接到SMCC连接子上，然后将抗体与PE-SMCC偶联。在第二种方法中，在R-PE分子上添加SH基团，而抗体连接到SPDP连接子上。然后，将抗体-SPDP分子与R-PE偶联。我们的结果表明，这两种偶联方法对抗体结合活性均无任何不利影响。
