Identification of Novel Single Chain Fragment Variable Antibodies Against TNF-alpha Using Phage Display Technology

Abstract

Purpose: Tumor necrosis factor alpha (TNF-alpha) is an inflammatory cytokine, involved in both physiological and pathological pathways. Because of central role of TNF-alpha in pathogenesis of inflammatory diseases, in the current study, we aimed to identify novel scFv antibodies against TNF-alpha using phage display technology. Methods: Using libraries composed of phagemid displaying scFv antibodies, four rounds of biopanning against TNF-alpha were carried out, which led to identification of scFvs capable of binding to TNF-alpha. The scFv antibody with appropriate binding affinity towards TNF-alpha, was amplified and used in ELISA experiment. Results: Titration of phage achieved from different rounds of biopanning showed an enrichment of specific anti-TNF-alpha phages during biopanning process. Using ELISA experiment, a binding constant (Kd) of 1.11 +/- 0.32 nM was determined for the phage displaying J48 scFv antibody. Conclusion: The findings in the current work revealed that the identified novel scFv antibody displayed at the N-terminal of minor coat proteins of phagemid binds TNF-alpha with suitable affinity. However, the soluble form of the antibody is needed to be produced and evaluated in more details regarding its binding properties to TNF-alpha.