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首页> 外文期刊>Biotechnology Progress >Factors Influencing Antibody Stability at Solid-Liquid Interfaces in a High Shear Environment
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Factors Influencing Antibody Stability at Solid-Liquid Interfaces in a High Shear Environment

机译：在高剪切环境中影响固液界面抗体稳定性的因素

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摘要

A rotating disk shear device was used to study the effect of interfacial shear on the structural integrity of human monoclonal antibodies of IgG4 isotype. Factors associated with the solution conditions (pH, ionic strength, surfactant concentration, temperature) and the interface (surface roughness) were studied for their effect on the rate of 1gG4monomer loss under high shear conditions. The structural integrity of the IgG4 was probed after exposure to interfacial shear effects by SDS-PAGE, IEF, dynamic light scattering, and peptide mapping by LC-MS. This analysis revealed that the main denaturation pathway of IgG4 exposed to these effects was the formation of large insoluble aggregates. Soluble aggregation, breakdown in primary structure, and chemical modifications were not detected. The dominant factors found to affect the rate of IgG4 monomer loss under interfacial shear conditions were found to be pH and the nanometer-scale surface roughness associated with the solid-liquid interface. Interestingly, temperature was not found to be a significant factor in the range tested (15-45°C). The addition of surfactant was found to have a significant stabilizing effect at concentrations up to 0.02% (w/v). Implications of these findings for the bioprocessing of this class of therapeutic protein are briefly discussed.
机译：使用旋转盘剪切装置研究界面剪切对IgG4同种型人单克隆抗体的结构完整性的影响。研究了在高剪切条件下与溶液条件（pH，离子强度，表面活性剂浓度，温度）和界面（表面粗糙度）有关的因素对1gG4单体损失速率的影响。在通过SDS-PAGE，IEF，动态光散射和LC-MS进行肽图分析暴露于界面剪切作用后，对IgG4的结构完整性进行了探测。该分析表明，暴露于这些作用的IgG4的主要变性途径是形成大的不溶性聚集体。未检测到可溶性聚集，一级结构破坏和化学修饰。发现在界面剪切条件下影响IgG4单体损失速率的主要因素是pH值和与固液界面相关的纳米级表面粗糙度。有趣的是，在所测试的范围内（15-45°C），温度不是重要因素。发现添加表面活性剂在浓度高达0.02％（w / v）时具有显着的稳定作用。简要讨论了这些发现对此类治疗蛋白的生物加工的影响。

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