Identification of novel single chain fragment variable antibodies against tnf-? using phage display technology

Abstract

Purpose: Tumor necrosis factor alpha (TNF-?) is an inflammatory cytokine, involved in both physiological and pathological pathways. Because of central role of TNF-? in pathogenesis of inflammatory diseases, in the current study, we aimed to identify novel scFv antibodies against TNF-? using phage display technology. Methods: Using libraries composed of phagemid displaying scFv antibodies, four rounds of biopanning against TNF-? were carried out, which led to identification of scFvs capable of binding to TNF-?. The scFv antibody with appropriate binding affinity towards TNF-?, was amplified and used in ELISA experiment. Results: Titration of phage achieved from different rounds of biopanning showed an enrichment of specific anti-TNF-? phages during biopanning process. Using ELISA experiment, a binding constant (Kd) of 1.11 آ± 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-? 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-?. é 2015 The Authors.