Biogen Idec, Inc, Cambridge, MA 02142, USA.
MAbs. 2009 Mar-Apr;1(2):128-41. doi: 10.4161/mabs.1.2.7631. Epub 2009 Mar 11.
Bispecific antibodies (BsAbs) represent an emerging class of biologics that achieve dual targeting with a single agent. Recombinant DNA technologies have facilitated a variety of creative bispecific designs with many promising therapeutic applications; however, practical methods for producing high quality BsAbs that have good product stability, long serum half-life, straightforward purification, and scalable production have largely been limiting. Here we describe a protein-engineering approach for producing stable, scalable tetravalent IgG-like BsAbs. The stability-engineered IgG-like BsAb was envisioned to target and crosslink two TNF family member receptors, TRAIL-R2 (TNF-Related Apoptosis Inducing Ligand Receptor-2) and LTbetaR (Lymphotoxin-beta Receptor), expressed on the surface of epithelial tumor cells with the goal of triggering an enhanced anti-tumor effect. Our IgG-like BsAbs consists of a stability-engineered anti-LTbetaR single chain Fv (scFv) genetically fused to either the N- or C-terminus of the heavy chain of a fulllength anti-TRAIL-R2 IgG1 monoclonal antibody. Both N- or C-terminal BsAbs were active in inhibiting tumor cell growth in vitro, and with some cell lines demonstrated enhanced activity relative to the combination of parental Abs. Pharmacokinetic studies in mice revealed long serum half-lives for the BsAbs. In murine tumor xenograft models, therapeutic treatment with the BsAbs resulted in reduction in tumor volume either comparable to or greater than the combination of parental antibodies, indicating that simultaneously targeting and cross-linking receptor pairs is an effective strategy for treating tumor cells. These studies support that stability-engineering is an enabling step for producing scalable IgG-like BsAbs with properties desirable for biopharmaceutical development.
双特异性抗体(BsAbs)是一类新兴的生物制剂,能够通过单个制剂实现双重靶向。重组 DNA 技术为多种具有广泛治疗应用前景的创新型双特异性设计提供了便利;然而,生产高质量、具有良好产品稳定性、长血清半衰期、简单纯化和可扩展生产的 BsAbs 的实用方法在很大程度上受到限制。在这里,我们描述了一种生产稳定、可扩展的四价 IgG 样 BsAbs 的蛋白质工程方法。设计稳定的 IgG 样 BsAb 的目的是靶向和交联两个 TNF 家族成员受体,即 TRAIL-R2(肿瘤坏死因子相关凋亡诱导配体受体-2)和 LTβR(淋巴毒素-β受体),这些受体表达在上皮肿瘤细胞表面,旨在触发增强的抗肿瘤作用。我们的 IgG 样 BsAbs 由经过稳定性工程改造的抗 LTβR 单链 Fv(scFv)通过基因融合到全长抗 TRAIL-R2 IgG1 单克隆抗体的重链的 N 端或 C 端组成。N 端或 C 端 BsAbs 在体外均能有效抑制肿瘤细胞生长,并且与某些细胞系相比,与亲本 Abs 的组合相比表现出增强的活性。在小鼠中的药代动力学研究显示 BsAbs 的血清半衰期较长。在小鼠肿瘤异种移植模型中,用 BsAbs 进行治疗可导致肿瘤体积减少,与亲本抗体的组合相当或大于该组合,表明同时靶向和交联受体对是治疗肿瘤细胞的有效策略。这些研究表明,稳定性工程是生产具有生物制药开发所需特性的可扩展 IgG 样 BsAbs 的必要步骤。
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