由于包括其速度和便利性的原因,皮下注射是许多抗体治疗剂的优选施用途径。然而,皮下给药的小体积限制(通常≤2mL)通常需要高浓度的抗体制剂(通常≥100mg/mL),这可能会导致物理化学问题。例如,具有大的疏水或带电荷的斑块的抗体可易于自相互作用,引起高粘度。这里,我们将X-射线晶体学与计算模型相结合,以预测抗胰高血糖素受体(GCGR)IgG1抗体易于发生自身相互作用的区域.对空间聚集倾向预测的疏水表面斑块中的互补决定区残基进行了广泛的突变分析,结合残留物级溶剂可及性,从分子动力学模拟得到的构象集合的平均值。动态光散射(DLS)用作~200种抗GCGRIgG1变体的自相互作用的中等通量筛选。在高浓度(180mg/mL)下测定的粘度与在低浓度(2-10mg/mL)下测定的DLS相互作用参数之间存在负相关。此外,抗GCGR变体很容易被鉴定为在亲本抗体的几倍内具有降低的粘度和抗原结合亲和力,对整体可开发性没有确定的影响。本文描述的方法可用于优化其他抗体以促进其高浓度的治疗性施用。
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.