title: 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.
abstract: Human tumour necrosis factor α (hTNFα) 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 hTNFα (sTNFα) as well as transmembrane hTNFα (tmTNFα) are more desirable hTNFα antagonists. Here, we report that novel anti-hTNFα human low-molecular-weight MAbs have been selected and identified using both sTNFα and tmTNFα 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 (VH /κ) provides an alternative promising molecular principle to generate biological agents for TNFα-dependent IMIDs. In this study, a panel of novel human VH /κs (F09, F21, F49 and F409) with high affinity (10(-10) -10(-9) mol/l) to neutralize sTNFα as well as tmTNFα 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 sTNFα as well as tmTNFα effectively, and both of them had great potential to inhibit hTNFα-mediated NF-κB activation. Soluble F21 and F409 were also able to inhibit the binding of hTNFα to TNFR1 and TNFR2. The new human antibodies described here have desirable capability to neutralize sTNFα as well as tmTNFα effectively with high affinity and reasonable stability; this may provide an alternative approach for patients who are not responding adequately to currently available anti-TNFα agents.
authors: Tianjin Institute of Health and Environmental Medicine, Tianjin, China.|New York University School of Medicine, New York, NY, USA.
authors: X-L
authors: XL
authors: Zhao
authors: 1st hospital of ShanXi Medical University, Taiyuan, China.
authors: L-F
authors: LF
authors: Tian
authors: 254th Hospital, Tianjin, China.
authors: S-J
authors: SJ
authors: Zhang
authors: 254th Hospital, Tianjin, China.
authors: J-M
authors: JM
authors: Li
authors: Tianjin College of Physical Education, Tianjin, China.
authors: H
authors: H
authors: Feng
authors: Tianjin Institute of Health and Environmental Medicine, Tianjin, China.
authors: L-M
authors: LM
authors: Wang
authors: Tianjin Institute of Health and Environmental Medicine, Tianjin, China.
authors: S
authors: S
authors: Wang
authors: Tianjin Institute of Health and Environmental Medicine, Tianjin, China.
authors: J
authors: J
authors: Wang
authors: Tianjin Institute of Health and Environmental Medicine, Tianjin, China.
authors: T
authors: T
authors: Wang
authors: Tianjin Institute of Health and Environmental Medicine, Tianjin, China.
authors: W-Q
authors: WQ
authors: Chen
mesh_terms: D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015723:Gene Library; D057809:HEK293 Cells; D006801:Humans; D007145:Immunoglobulin kappa-Chains / Q000235:genetics / Q000276:immunology*; D015202:Protein Engineering / Q000379:methods*; D017434:Protein Structure, Tertiary; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000378:metabolism; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000378:metabolism; D012270:Ribosomes / Q000235:genetics; D061905:Single-Domain Antibodies / Q000235:genetics / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D007145:Immunoglobulin kappa-Chains; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D061905:Single-Domain Antibodies; C439524:TNF protein, human; D014409:Tumor Necrosis Factor-alpha