Novel Human Three-Domain Antibody Fragments Against sTNFα as Well as tmTNFα with High Affinity Generated by the Combination of Ribosome Display and E.coli Expression System

作者：Zhao, X. -L.; Tian, L. -F.; Zhang, S. -J.; Li, J. -M.; Feng, H.; Wang, L. -M.; Wang, S.; Wang, J.; Wang, T.; Chen, W. -Q.^*

来源：Scandinavian Journal of Immunology, 2016, 83(4): 267-278.

DOI：10.1111/sji.12417

• TUMOR-NECROSIS-FACTOR
• RHEUMATOID-ARTHRITIS
• IN-VITRO
• MONOCLONAL-ANTIBODY
• ESCHERICHIA-COLI
• TNF-ALPHA
• CONCOMITANT METHOTREXATE
• CERTOLIZUMAB PEGOL
• CLINICAL-EFFICACY
• BINDING

Human tumour necrosis factor (hTNF) has been proved to be a validated therapeutic target in a number of immune-mediated inflammatory diseases (IMIDs). Fully human monoclonal antibodies (mAbs) that can neutralize soluble hTNF (sTNF) as well as transmembrane hTNF (tmTNF) are more desirable hTNF antagonists. Here, we report that novel anti-hTNF human low-molecular-weight MAbs have been selected and identified using both sTNF and tmTNF as target antigens by the combination of ribosome display and E.coli expression system for the first time. As a newly born engineering small molecular antibody, three-domain antibody fragment (V-H/) provides an alternative promising molecular principle to generate biological agents for TNF-dependent IMIDs. In this study, a panel of novel human V-H/s (F09, F21, F49 and F409) with high affinity (10(-10)-10(-9)mol/l) to neutralize sTNF as well as tmTNF was generated by the combination of ribosome display and E.coli expression system. Among the four clones, F21 and F409 could reduce cytotoxicity on L929 cells induced by sTNF as well as tmTNF effectively, and both of them had great potential to inhibit hTNF-mediated NF-B activation. Soluble F21 and F409 were also able to inhibit the binding of hTNF to TNFR1 and TNFR2. The new human antibodies described here have desirable capability to neutralize sTNF as well as tmTNF effectively with high affinity and reasonable stability; this may provide an alternative approach for patients who are not responding adequately to currently available anti-TNF agents.

• 出版日期2016-4
• 单位山西医科大学