Characterization of Novel Fragment Antibodies Against TNF-alpha Isolated Using Phage Display Technique
| Author | Ali Akbar Alizadeha | en |
| Author | Maryam Hamzeh-Mivehroud | en |
| Author | Elnaz Haddad | en |
| Author | Nazanin Haddad | en |
| Author | Mehdi Sharifi | en |
| Author | Samin Mohammadi | en |
| Author | Samira Pourtaghi-anvarian | en |
| Author | Siavoush Dastmalchi | en |
| Issued Date | 2019-04-30 | en |
| Abstract | Tumor necrosis factor alpha (TNF-α) is an inflammatory cytokine which plays crucial roles in pathogenesis of inflammatory diseases. The current study aimed to investigate the binding abilities of I44 and I49 domain antibodies to TNF-α. The dAbs were expressed in bacterial expression system and purified by affinity chromatography using Ni-sepharose column. The expression and purity of the proteins were evaluated using western blotting and SDS-PAGE techniques, respectively. ELISA experiment showed that I44 and I49 dAbs bind to TNF-α with the binding constants (Kd) of 5.18 ± 1.41 and 2.42 ± 0.55 µM, respectively. The inhibitory effect of dAbs on TNF-α biological effect was determined in MTT assay in which I44 and I49 prevented TNF-α cell cytotoxicity with IC50 values of 6.61 and 3.64 µM, respectively. The identified anti-TNF-α dAbs could bind to and inhibit TNF-α activity. The dAbs activities can be attributed to their ability to establish hydrogen bonds as well as hydrophobic contacts with TNF-α. The results of the current study can pave the way for further structural studies in order to introduce new more potent anti-TNF-α antibodies. | en |
| DOI | https://doi.org/10.22037/ijpr.2019.1100646 | en |
| Keyword | TNF-α | en |
| Keyword | Domain antibodies | en |
| Keyword | Recombinant protein production | en |
| Keyword | Molecular docking | en |
| Keyword | Phage display | en |
| Publisher | Brieflands | en |
| Title | Characterization of Novel Fragment Antibodies Against TNF-alpha Isolated Using Phage Display Technique | en |
| Type | Original Article | en |