文献类型：期刊文献

英文题名：Designing and constructing a phage display synthesized single domain antibodies library based on camel VHHs frame for screening and identifying humanized TNF-α-specific nanobody

作者：Nie, Jifan[1];Ma, Xingyuan[1];Hu, Fabiao[1];Miao, Hui[1];Feng, Xin[1];Zhang, Peiwen[2];Han, Myong Hun[1,3];You, Fang[4];Yang, Yi[4,5];Zhang, Wenlian[1,6];Zheng, Wenyun[2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, Shanghai Key Lab New Drug, Sch Pharm, Shanghai 200237, Peoples R China;[3]Kim Il Sung Univ, Dept Genet, Fac Life Sci, Pyongyang 999093, North Korea;[4]Natl Univ Singapore, Dept Chem & Biomol Engn, Singapore 117585, Singapore;[5]SinGENE Biotech Pte Ltd, Singapore Sci Pk, Singapore 118258, Singapore;[6]Univ North Carolina, Ctr Translat Biomed Res, Greensboro, NC 27310 USA

年份：2021

卷号：137

外文期刊名：BIOMEDICINE & PHARMACOTHERAPY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000634759700005)】；

基金：This study was supported by the National Key Research and Development Project of China (2018YFA0902804) , the National Natural Science Foundation (31670944, 81673345) , and the Science and Technology Innovation Action Plan of Shanghai (17431904600) .

语种：英文

外文关键词：Single domain antibodies; Phage display library; Designing and constructing; Humanized TNF-alpha-specific Nb; Screening and identifyin

摘要：Tumor necrosis factor (TNF-alpha) is an important clinically tested cytokine that could induce autoimmune diseases and inflammation. Therefore, the anti-TNF-alpha therapy strategy was developed and used therapeutically in various diseases, especially in the cytokine storm associated chimeric antigen receptor (CAR) T-cell therapy and antiviral therapy. Compare with other anti-TNF-alpha inhibitors, anti-TNF-alpha Nb (nanobody) has many unique advantages. Herein, we reported a novel humanized scaffold for library construction, which could be soluble and expressed in Escherichia coli (E.coli), and the efficiency capacity could reach as high as 2.01 x 10(9). Meanwhile, an anti-TNF-alpha Nb was selected for further study after 4 rounds of screening, NT-3, as the optimal Nb could effectively inhibit TNF-mediated cytotoxicity. The IC50 of NT-3 was determined as 0.804 mu M, and its apoptosis inhibition rate was 62.47 % in L929 cells. Furthermore, the molecular docking results showed that complementarity-determining regions (CDRs) of NT-3 could connect to TNF for blocking function through strong hydrogen bonds and salt bridges. In general, our study not only provided a good Nb screening platform in vitro without animal immunization, but also generated a series of novel humanized anti-TNF-alpha Nb candidates with potential applications.