Engineering a therapeutic IgG molecule to address cysteinylation, aggregation and enhance thermal stability and expression.
Engineering a therapeutic IgG molecule to address cysteinylation, aggregation and enhance thermal stability and expression.
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作者：
Buchanan A;Clementel V;Woods R;Harn N;Bowen MA;Mo W;Popovic B;Bishop SM;Dall'Acqua W;Minter R;Jermutus L;Bedian V
抗体可发生多种共价和非共价降解反应，对效力、安全性、生产和储存产生不利影响。我们已经鉴定了抗血管生成素2（Ang 2 mAb）的抗体，其在体外中和Ang 2与其受体的结合并在体内抑制肿瘤生长。尽管具有良好的药理学活性，但Ang 2 mAb制剂是异质的，快速聚集且表达较差。在此，我们报道了抗体可变和恒定结构域的工程化以产生具有降低的聚集倾向、增强的均一性、11°C升高的Tm、26倍提高的表达水平和保留的活性的抗体。工程分子MEDI-3617现在与临床试验所需的大规模材料供应兼容，目前正在癌症患者中进行第1阶段评估。这是第一份报告，以描述解决非典型的半胱氨酸残基的治疗性抗体的稳定性工程和设计策略，这里报道的一般适用于其他治疗性抗体和蛋白质。
Antibodies can undergo a variety of covalent and non-covalent degradation reactions that have adverse effects on efficacy, safety, manufacture and storage. We had identified an antibody to Angiopoietin 2 (Ang2 mAb) that neutralizes Ang2 binding to its receptor in vitro and inhibits tumor growth in vivo. Despite favorable pharmacological activity, the Ang2 mAb preparations were heterogeneous, aggregated rapidly and were poorly expressed. Here, we report the engineering of the antibody variable and constant domains to generate an antibody with reduced propensity to aggregate, enhanced homogeneity, 11°C elevated Tm, 26-fold improved level of expression and retained activity. The engineered molecule, MEDI-3617, is now compatible with the large scale material supply required for clinical trials and is currently being evaluated in Phase 1 in cancer patients. This is the first report to describe the stability engineering of a therapeutic antibody addressing non canonical cysteine residues and the design strategy reported here is generally applicable to other therapeutic antibodies and proteins.