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| Comparison of the Inhibitory Efficiency of M13 Based 7-mer and 12-mer Phage Display Libraries Derived Peptides as Tumor Necrosis Factor Alpha Antagonist |
| HU Chang-wu, XIE Jun, ZHU Nai-shuo |
| State Key Lab of Genetic Engineering, Lad of Molecular Immunology, School of Life Science, Fudan University, Shanghai 200438, China |
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Abstract The antagonist of tumor necrosis factor alpha is the first choice for the treatment of multiple inflammatory autoimmune diseases. Treatment with TNFα antibodies is restricted by their side effects, particularly, the production of anti-antibodies, which seriously affect the treatment efficacy and drug metabolism. Short peptides have low immunogenicity, and compared to small molecules, they also have lower toxicity and stronger target specificity. Using M13 based 7-mer and 12-mer peptides phage display libraries to screen TNFα binding peptide ligands, and analyzed the affinity and functionality of the selected TNFα antagonist. After 3-4 rounds of screening, two 7-mer and two 12-mer peptide sequences were obtained. Binding affinity of the synthetic peptides for TNFα was determined by ELISA. 7-mer peptide with number 632 showed affinity of Kd=138nmol/L, while the 12-mer peptide with number 636 showed lower affinity of Kd=8.59μmol/L. Insight II software was used to carry out Zdock with TNFα dimer, and it was found that both the 7-mer peptide could bind to TNFα with a more stable state than the 12-mer peptide. At the cellular level, the peptide 632 were more resistant to the activity of TNFα than peptide 636. In the presence of peptide 632, the survival rate of L929 cells induced by TNFα was 3 times higher, but the 636 peptides were only 2 times. Altogether, the 7-mer peptides were more suitable than 12-mer peptide as TNFα antagonist.
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Received: 01 November 2016
Published: 25 May 2017
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