Freezing-induced perturbation of tertiary structure of a monoclonal antibody.
Freezing-induced perturbation of tertiary structure of a monoclonal antibody.
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DOI：
10.1002/jps.24013
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发表时间：
2014-07
影响因子：
3.8
通讯作者：
Carpenter, John F.
中科院分区：
文献类型：
--
作者：
Liu, Lu;Braun, Latoya Jones;Wang, Wei;Randolph, Theodore W.;Carpenter, John F.
关键词：
我们用内源色氨酸荧光光谱研究了pH和溶液添加剂对冷冻诱导的单抗三级结构扰动的影响。一般而言，冷冻引起单抗三级结构的扰动，这取决于制剂中存在的pH或赋形剂的可逆或不可逆。在冻融样品中发生了蛋白质聚集，其中观察到了三级结构的扰动，但形成的蛋白质聚集体的水平与结构扰动的程度不成比例。在冻融样品中也发生了蛋白质聚集，没有明显的结构扰动，这很可能是因为蛋白质和盐的冷冻浓缩，从而降低了蛋白质胶体的稳定性。因此，冷冻诱导的蛋白质聚集可能会也可能不会首先涉及其天然结构的扰动，随后是形成聚集体的组装过程。根据解决方案的条件，这两个步骤都可以是速率限制。最后，这项研究证明了荧光光谱作为筛选冻融压力下的治疗性蛋白质制剂的有价值的工具的潜力。
We studied the effects of pH and solution additives on freezing-induced perturbations in the tertiary structure of a monoclonal antibody (mAb) by intrinsic tryptophan fluorescence spectroscopy. In general, freezing caused perturbations in the tertiary structure of the mAb, which were reversible or irreversible depending on the pH or excipients present in the formulation. Protein aggregation occurred in freeze–thawed samples in which perturbations of the tertiary structure were observed, but the levels of protein aggregates formed were not proportional to the degree of structural perturbation. Protein aggregation also occurred in freeze–thawed samples without obvious structural perturbations, most likely because of freeze concentration of protein and salts, and thus reduced protein colloidal stability. Therefore, freezing-induced protein aggregation may or may not first involve the perturbation of its native structure, followed by the assembly processes to form aggregates. Depending on the solution conditions, either step can be rate limiting. Finally, this study demonstrates the potential of fluorescence spectroscopy as a valuable tool for screening therapeutic protein formulations subjected to freeze–thaw stress.
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影响因子：
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