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Updated: Mar 26, 2026
Bacterial Inner-membrane Display for Screening a Library of Antibody Fragments
Published on: October 15, 2016
Identification of Novel Single Chain Fragment Variable Antibodies Against TNF-α Using Phage Display Technology
Ali Akbar Alizadeh1, Maryam Hamzeh-Mivehroud2, Siavoush Dastmalchi3
1Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. ; Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran. ; Students Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran. ; School of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
Purpose:
Tumor necrosis factor alpha (TNF-α) is an inflammatory cytokine, involved in both physiological and pathological pathways. Because of central role of TNF-α in pathogenesis of inflammatory diseases, in the current study, we aimed to identify novel scFv antibodies against TNF-α using phage display technology.
Methods:
Using libraries composed of phagemid displaying scFv antibodies, four rounds of biopanning against TNF-α were carried out, which led to identification of scFvs capable of binding to TNF-α. The scFv antibody with appropriate binding affinity towards TNF-α, was amplified and used in ELISA experiment.
Results:
Titration of phage achieved from different rounds of biopanning showed an enrichment of specific anti-TNF-α 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-α 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-α.
