Tabriz University of Medical Sciences Advanced Pharmaceutical Bulletin 2228-5881 5 5 2015 12 31 Production and Purification of a Novel Anti-TNF-α Single Chain Fragment Variable Antibody 667 672 10.15171/apb.2015.091 EN Ali Akbar Alizadeh Maryam Hamzeh-Mivehroud Siavoush Dastmalchi Journal Article 10.15171/apb.2015.091 2015 10 19 2015 11 01 Purpose: TNF-α is an inflammatory cytokine with a key role in initiation of inflammatory responses. Anti-TNF-α antibodies are being used in clinic for the purpose of diagnosis and treatment due to their high specificity. The objective of the current study was to express and purify an anti-TNF-α scFv antibody identified by phage display technology. Methods: The DNA coding sequence of the identified scFv was cloned into pET28a vector and the corresponding protein was expressed as 6×His tagged using E.coli BL21 (DE3) pLysS expression system followed by affinity purification on Ni-Sepharose affinity column. Results: The J44 scFv antibody was cloned into the expression vector and successfully expressed and purified. The purity of the scFv fraction was confirmed using SDS-PAGE analysis. Western blotting technique was used to detect expression of 6×His tagged protein. Conclusion: In the current study an anti-TNF-α scFv antibody was successfully expressed in bacterial expression system and purified on affinity column. The purified protein can be used in different in vitro and in vivo experiments in order to elucidate its functionality.