On the role of salt type and concentration on the stability behavior of a monoclonal antibody solution
On the role of salt type and concentration on the stability behavior of a monoclonal antibody solution
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DOI：
10.1016/j.bpc.2012.05.004
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发表时间：
2012-07-01
影响因子：
3.8
通讯作者：
Morbidelli, Massimo
中科院分区：
文献类型：
--
作者：
Arosio, Paolo;Jaquet, Baptiste;Morbidelli, Massimo
蛋白质和盐的相互作用调节蛋白质的溶解度和稳定性，特别是一些蛋白质相关的过程，如盐析和聚集。以IgG2单克隆抗体为模型多结构域治疗蛋白，研究了盐对小聚集数蛋白簇可逆形成的影响。低聚物的形成已被定量的大小排斥色谱（SEC）。发现盐效应具有很强的离子特异性和pH依赖性。特别是，在pH 3.0时，只有阴离子影响聚集倾向，而在pH 4.0时，阴离子和阳离子都影响聚集速率。除硫酸盐外，阴离子效应的排名遵循Hofmeister系列，而阳离子效应的排名则不遵循Hofmeister系列。此外，观察到作为盐浓度函数的聚集倾向的最大值（即存在再稳定）。通过将聚集动力学与蛋白质结构和表面能的实验研究相关联，表明pH和盐浓度的变化不仅通过电荷筛选和各种溶剂化力诱导聚集，而且通过形成具有部分有序结构和一定程度疏水性的蛋白质中间体诱导聚集。溶剂化力与盐诱导的蛋白质二级结构之间的复杂相互作用解释了观察到的实验结果，这些结果与大盐浓度下的再稳定、离子特异性和硫酸盐阴离子的特殊行为有关。(c) 2012 Elsevier B.V.版权所有
Protein salt interactions regulate protein solubility and stability and in particular several protein related processes, such as salting-out and aggregation. Using an IgG2 monoclonal antibody as a model multi-domain therapeutic protein, we have investigated the salt effect on the reversible formation of protein clusters with small aggregation number. The oligomer formation has been quantified by size exclusion chromatography (SEC). It is found that the salt effect is strongly ion specific and pH dependent. In particular, at pH 3.0 only anions affect the aggregation propensity, while at pH 4.0 both anions and cations influence the aggregation rate. The ranking of the anion effect follows the Hofmeister series with the only exception of sulfate, while that of the cation effect does not. In addition, a maximum of the aggregation propensity as a function of salt concentration is observed (i.e., presence of re-stabilization).By correlating the aggregation kinetics to the experimental investigation of protein structure and surface energy, it is shown that changes in pH and salt concentration induce aggregation not only through charge screening and various solvation forces, but also through the formation of protein intermediates characterized by partially ordered structures and certain degrees of hydrophobicity. The complex interaction between the solvation forces and such protein secondary structures induced by salts explains the observed experimental results relative to re-stabilization at large salt concentrations, ion specificity and the peculiar behavior of the sulfate anion. (c) 2012 Elsevier B.V. All rights reserved.