Center for Infection and Immunity Research, State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Wuhan, 430062, Hubei, People's Republic of China.
Division of Rheumatology, Department of Internal Medicine, University of Texas Health Science Center at Houston, Houston, TX, 77030, USA.
Biotechnol Lett. 2020 Apr;42(4):557-569. doi: 10.1007/s10529-020-02830-3. Epub 2020 Feb 10.
To provide an alternative therapeutic modality for rheumatoid arthritis (RA), a novel bispecific antibody (BsAb) targeting human tumor necrosis factor α (TNF-α) and human complement component C5a was constructed.
BsAb was expressed in Pichia pastoris and secreted into the culture medium as a functional protein. In vitro functional study demonstrated that BsAb could simultaneously bind to TNF-α and C5a and neutralize their biological actions. Furthermore, BsAb showed significant improvements in both the antigen-binding affinity and the neutralizing ability as compared to its original antibodies produced in E. coli. It was also found that TNF-α and C5a had an additive/synergistic effect on promoting the production of inflammatory cytokines and chemokines and C5a receptor (C5aR) expression in human macrophages. Compared to single inhibition of TNF-α or C5a with respective antibody, BsAb showed a superior efficacy in blocking inflammatory cytokines, chemokines, and C5aR response, as well as in lowering the C5a-mediated chemotaxis of macrophages via C5aR in vitro.
With improved production processing and the ability to simultaneously block TNF-α and C5a action, BsAb has a great potential to be developed into a therapeutic agent and may offer a better therapeutic index for RA.
为类风湿关节炎（RA）提供一种新的治疗方法，构建了一种靶向人肿瘤坏死因子α（TNF-α）和人补体成分 C5a 的新型双特异性抗体（BsAb）。
BsAb 在毕赤酵母中表达并分泌到培养基中作为一种功能性蛋白。体外功能研究表明，BsAb 可以同时结合 TNF-α 和 C5a 并中和它们的生物学作用。此外，与在大肠杆菌中产生的原始抗体相比，BsAb 在抗原结合亲和力和中和能力方面都有显著提高。还发现 TNF-α 和 C5a 在促进人巨噬细胞产生炎症细胞因子和趋化因子以及 C5a 受体（C5aR）表达方面具有相加/协同作用。与单独抑制 TNF-α或 C5a 相比，BsAb 在阻断炎症细胞因子、趋化因子和 C5aR 反应以及降低 C5a 介导的巨噬细胞趋化性方面具有更好的疗效通过体外 C5aR。
BsAb 具有改进的生产工艺和同时阻断 TNF-α和 C5a 作用的能力，有很大潜力开发成治疗剂，并可能为 RA 提供更好的治疗指数。
