%0 Journal Article
%T Affinity Purification of Tumor Necrosis Factor-¦Á Expressed in Raji Cells by Produced scFv Antibody Coupled CNBr-Activated Sepharose
%A Jalal Abdolalizadeh
%A Jafar Majidi Zolbanin
%A Mohammad Nouri
%A Behzad Baradaran
%J Advanced Pharmaceutical Bulletin
%D 2013
%I Tabriz University of Medical Sciences
%R 10.5681/apb.2013.004
%X Purpose: Recombinant tumor necrosis factor-alpha (TNF-¦Á) has been utilized as an antineoplastic agent for the treatment of patients with melanoma and sarcoma. It targets tumor cell antigens by impressing tumor-associated vessels. Protein purification with affinity chromatography has been widely used in the downstream processing of pharmaceutical-grade proteins. Methods: In this study, we examined the potential of our produced anti-TNF-  scFv fragments for purification of TNF-¦Á produced by Raji cells.   he Raji cells were induced by lipopolysaccharides (LPS) to express TNF-¦Á. Western blotting and Fluorescence-activated cell sorting (FACS) flow cytometry analyses were used to evaluate the TNF-¦Á expression. The anti-TNF-¦Á scFv selected from antibody phage display library was coupled to CNBr-activated sepharose 4B beads used for affinity purification of expressed TNF-¦Á and the purity of the protein was assessed by SDS-PAGE. Results: Western blot and FACS flow cytometry analyses showed the successful expression of TNF-¦Á with Raji cells. SDS-PAGE analysis showed the performance of scFv for purification of TNF-¦Á protein with purity over 95%. Conclusion: These findings confirm not only the potential of the produced scFv antibody fragments but also this highly pure recombinant TNF-¦Á protein can be applied for various in vitro and in vivo applications.
%K TNF-¦Á expression
%K Affinity Purification
%K Monoclonal antibody
%K LPS
%U http://journals.tbzmed.ac.ir/PDF/APB/Manuscript/APB-3-19.pdf