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Advanced Pharmaceutical Bulletin
ISSN: 2228-5881      eISSN: 2251-7308  
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Article History
Submitted: 19 Oct 2015
Revised: 31 Oct 2015
Accepted: 01 Nov 2015
First published online: 31 Dec 2015

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Adv Pharm Bull. 2015;5(5):661-666 doi: 10.15171/apb.2015.090
PMID:26793613        PMCID:PMC4708038

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

Original Research

Ali Akbar Alizadeh 1,2,3,4, Maryam Hamzeh-Mivehroud 5,2,4, Siavoush Dastmalchi 2,4 *

1 Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
2 Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
3 Students Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
4 School of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
5 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.



Abstract
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.





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