Variable domain mutational analysis to probe the molecular mechanisms of high viscosity of an IgG1 antibody.

IF 5.6 2区 医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL
mAbs Pub Date : 2024-01-01 Epub Date: 2024-01-25 DOI:10.1080/19420862.2024.2304282
Jing Dai, Saeed Izadi, Jonathan Zarzar, Patrick Wu, Angela Oh, Paul J Carter
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引用次数: 0

Abstract

Subcutaneous injection is the preferred route of administration for many antibody therapeutics for reasons that include its speed and convenience. However, the small volume limit (typically 2 mL) for subcutaneous delivery often necessitates antibody formulations at high concentrations (commonly ≥100 mg/mL), which may lead to physicochemical problems. For example, antibodies with large hydrophobic or charged patches can be prone to self-interaction giving rise to high viscosity. Here, we combined X-ray crystallography with computational modeling to predict regions of an anti-glucagon receptor (GCGR) IgG1 antibody prone to self-interaction. An extensive mutational analysis was undertaken of the complementarity-determining region residues residing in hydrophobic surface patches predicted by spatial aggregation propensity, in conjunction with residue-level solvent accessibility, averaged over conformational ensembles from molecular dynamics simulations. Dynamic light scattering (DLS) was used as a medium throughput screen for self-interaction of ~ 200 anti-GCGR IgG1 variants. A negative correlation was found between the viscosity determined at high concentration (180 mg/mL) and the DLS interaction parameter measured at low concentration (2-10 mg/mL). Additionally, anti-GCGR variants were readily identified with reduced viscosity and antigen-binding affinity within a few fold of the parent antibody, with no identified impact on overall developability. The methods described here may be useful in the optimization of other antibodies to facilitate their therapeutic administration at high concentration.

通过变异结构域突变分析探究 IgG1 抗体高粘度的分子机制。
皮下注射是许多抗体疗法的首选给药途径,原因包括快捷方便。然而,由于皮下注射的容量限制较小(通常≤2 mL),因此通常需要高浓度(通常≥100 mg/mL)的抗体配方,这可能会导致理化问题。例如,具有大面积疏水或带电斑块的抗体容易发生自相互作用,从而导致高粘度。在这里,我们将 X 射线晶体学与计算建模相结合,预测了抗胰高血糖素受体(GCGR)IgG1 抗体中容易发生自相互作用的区域。通过分子动力学模拟构象组合的平均值,结合空间聚集倾向预测的残基级溶剂可及性,对位于疏水表面斑块的互补性决定区残基进行了广泛的突变分析。利用动态光散射(DLS)对约 200 种抗-GCGR IgG1 变体的自相互作用进行了中等通量筛选。结果发现,在高浓度(180 毫克/毫升)下测定的粘度与在低浓度(2-10 毫克/毫升)下测定的 DLS 相互作用参数之间存在负相关。此外,抗 GCGR 变体很容易鉴定出来,其粘度和抗原结合亲和力比母体抗体降低了几倍,但未发现对整体开发性有影响。本文所述的方法可能有助于优化其他抗体,以促进其在高浓度下的治疗用药。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
mAbs
mAbs 工程技术-仪器仪表
CiteScore
10.70
自引率
11.30%
发文量
77
审稿时长
6-12 weeks
期刊介绍: mAbs is a multi-disciplinary journal dedicated to the art and science of antibody research and development. The journal has a strong scientific and medical focus, but also strives to serve a broader readership. The articles are thus of interest to scientists, clinical researchers, and physicians, as well as the wider mAb community, including our readers involved in technology transfer, legal issues, investment, strategic planning and the regulation of therapeutics.
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