Classification and Characterization of Therapeutic Antibody Aggregates
Classification and Characterization of Therapeutic Antibody Aggregates
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DOI：
10.1074/jbc.m110.160457
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发表时间：
2011-07-15
影响因子：
4.8
通讯作者：
Narhi, Linda O.
中科院分区：
文献类型：
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作者：
Joubert, Marisa K.;Luo, Quanzhou;Narhi, Linda O.
许多不同的应激技术被应用于一种单抗(Ig G(2))，以产生具有不同属性和形态的蛋白质颗粒。对聚合溶液的聚集率、颗粒计数、粒度分布、形态、二级和三级结构的变化、表面疏水性、金属含量和可逆性进行了评估。另一份报告(Luo，Q.，Joubert，M.K.，Stevenson，R.，Narhi，L.O.和Wypych，J.(2011)J.Biol)中也发现了化学修饰。化学。286、25134-25144)。根据所描述的特征，集合体被分为七个不同的类别。强调了几个额外的分子(来自免疫球蛋白(1)和免疫球蛋白(2)亚型以及静脉注射免疫球蛋白)，发现它们被定义为相同的分类系统。蛋白质聚集的机制和形成的聚集类型取决于所施加的胁迫的性质。在相同的压力下，不同的免疫球蛋白分子似乎以相似的机制聚集在一起。在严酷的机械应力作用下形成的团聚体显示出最多的亚可见颗粒，而在热应力作用下产生的团聚体显示出最多的可见颗粒。大多数班级表现出高阶结构的破坏，无序程度取决于应激过程。在pH为5的缓冲液中稀释后，所有类别的颗粒(热应力除外)至少部分可逆。在分离的金属催化的聚集体中检测到高铜含量，这种压力以前被证明会产生免疫原性聚集体。总而言之，蛋白质聚集体可能是一个非常不同的群体，他们的品质是所经历的压力类型的结果。
A host of diverse stress techniques was applied to a monoclonal antibody (IgG(2)) to yield protein particles with varying attributes and morphologies. Aggregated solutions were evaluated for percent aggregation, particle counts, size distribution, morphology, changes in secondary and tertiary structure, surface hydrophobicity, metal content, and reversibility. Chemical modifications were also identified in a separate report (Luo, Q., Joubert, M. K., Stevenson, R., Narhi, L. O., and Wypych, J. (2011) J. Biol. Chem. 286, 25134-25144). Aggregates were categorized into seven discrete classes, based on the traits described. Several additional molecules (from the IgG(1) and IgG(2) subtypes as well as intravenous IgG) were stressed and found to be defined with the same classification system. The mechanism of protein aggregation and the type of aggregate formed depends on the nature of the stress applied. Different IgG molecules appear to aggregate by a similar mechanism under the same applied stress. Aggregates created by harsh mechanical stress showed the largest number of subvisible particles, and the class generated by thermal stress displayed the largest number of visible particles. Most classes showed a disruption of the higher order structure, with the degree of disorder depending on the stress process. Particles in all classes (except thermal stress) were at least partially reversible upon dilution in pH 5 buffer. High copper content was detected in isolated metal-catalyzed aggregates, a stress previously shown to produce immunogenic aggregates. In conclusion, protein aggregates can be a very heterogeneous population, whose qualities are the result of the type of stress that was experienced.