Identification and characterization of a monoclonal antibody variant species with a clipping in the complementarity determining region isolated by size exclusion chromatography under native conditions.
Identification and characterization of a monoclonal antibody variant species with a clipping in the complementarity determining region isolated by size exclusion chromatography under native conditions.
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通过在天然条件下通过尺寸排阻色谱法分离的互补决定区中的剪辑来鉴定和表征单克隆抗体变体种类。
DOI：
10.1016/j.xphs.2021.05.023
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发表时间：
2021
影响因子：
3.8
通讯作者：
K. Wakamatsu
中科院分区：
文献类型：
--
作者：
Yuriko Atsumi;Natsuko Sakurai;K. Nishimura;Katsuyoshi Yamazaki;K. Wakamatsu
单克隆抗体（mAb）片段的含量是药品单克隆抗体产品的重要质量指标，必须对其进行安全控制。单克隆抗体铰链区的肽键容易被水解，产生Fc-Fab片段，其有效性低于完整抗体。在天然条件下，Fc-Fab片段可以通过大小排斥色谱（SEC）从完整的抗体分子中分离出来。虽然一个片段在互补决定区（CDR）中产生了几个片段，但它们的功效尚未得到分析。这是因为这些片段由于分子大小相似，在天然条件下无法从完整的抗体中分离出来。在这里，我们报告了在CDR中剪切的贝伐单抗变体，其产生的片段在变体中保持完整，可以通过选择适当的SEC柱在自然条件下分离。
The content of monoclonal antibody (mAb) fragments in pharmaceutical mAb products is a critical quality attribute and should be controlled for safety. Peptide bonds in the hinge region of mAbs are susceptible to hydrolysis, generating Fc-Fab fragments, which are associated with lower efficacy than the intact antibody. Fc-Fab fragments can be separated from intact antibody molecules under native conditions by size exclusion chromatography (SEC). Although several fragments generated by a clip in the complementarity determining region (CDR) have been reported, their efficacies have not been analyzed. This is because these fragments could not be separated from intact antibodies under native conditions owing to their similar molecular sizes. Here, we report that bevacizumab variant with clipping in the CDR, with the resulting fragments remaining intact in the variant, can be isolated under native conditions by selecting an adequate SEC column.