Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, MD, USA.
Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
MAbs. 2022 Jan-Dec;14(1):2095704. doi: 10.1080/19420862.2022.2095704.
Manipulation of glycosylation patterns, i.e., glycoengineering, is incorporated in the therapeutic antibody development workflow to ensure clinical safety, and this approach has also been used to modulate the biological activities, functions, or pharmacological properties of antibody drugs. Whereas most existing glycoengineering strategies focus on the canonical glycans found in the constant domain of immunoglobulin G (IgG) antibodies, we report a new strategy to leverage the untapped potential of atypical glycosylation patterns in the variable domains, which naturally occur in 15% to 25% of IgG antibodies. Glycosylation sites were added to the antigen-binding regions of two functionally divergent, interleukin-2-binding monoclonal antibodies. We used computational tools to rationally install various N-glycosylation consensus sequences into the antibody variable domains, creating "glycovariants" of these molecules. Strikingly, almost all the glycovariants were successfully glycosylated at their newly installed N-glycan sites, without reduction of the antibody's native function. Importantly, certain glycovariants exhibited modified activities compared to the parent antibody, showing the potential of our glycoengineering strategy to modulate biological function of antibodies involved in multi-component receptor systems. Finally, when coupled with a high-flux sialic acid precursor, a glycovariant with two installed glycosylation sites demonstrated superior in vivo half-life. Collectively, these findings validate a versatile glycoengineering strategy that introduces atypical glycosylation into therapeutic antibodies in order to improve their efficacy and, in certain instances, modulate their activity early in the drug development process.
糖基化模式的调控，即糖基工程，被纳入治疗性抗体的开发工作流程中，以确保临床安全性，这种方法也被用于调节抗体药物的生物学活性、功能或药理学性质。虽然大多数现有的糖基工程策略都集中在免疫球蛋白 G（IgG）抗体的恒定域中发现的典型聚糖上，但我们报告了一种利用可变域中未开发的非典型糖基化模式的新策略，这些模式在 15%到 25%的 IgG 抗体中自然发生。在两种功能不同的白细胞介素-2 结合单克隆抗体的抗原结合区添加了糖基化位点。我们使用计算工具将各种 N-糖基化共识序列合理地安装到抗体的可变域中，从而产生这些分子的“糖变体”。引人注目的是，几乎所有的糖变体都成功地在其新安装的 N-聚糖位点上进行了糖基化，而不会降低抗体的天然功能。重要的是，某些糖变体与亲本抗体相比表现出不同的活性，表明我们的糖基工程策略具有调节多组分受体系统中参与的抗体的生物学功能的潜力。最后，当与高通量唾液酸前体偶联时，具有两个安装的糖基化位点的糖变体表现出优越的体内半衰期。总之，这些发现验证了一种通用的糖基工程策略，该策略将非典型糖基化引入治疗性抗体中，以提高其疗效，并在药物开发过程的早期在某些情况下调节其活性。
