Assessment of Physical Stability of an Antibody Drug Conjugate by Higher Order Structure Analysis: Impact of Thiol- Maleimide Chemistry
Assessment of Physical Stability of an Antibody Drug Conjugate by Higher Order Structure Analysis: Impact of Thiol- Maleimide Chemistry
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DOI：
10.1007/s11095-013-1274-2
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发表时间：
2014-07-01
影响因子：
3.7
通讯作者：
Singh, Satish K.
中科院分区：
文献类型：
--
作者：
Guo, Jianxin;Kumar, Sandeep;Singh, Satish K.
为了提供系统的生物物理方法，以更好地理解缀合化学对抗体药物缀合物（ADC）的高级结构和物理稳定性的影响，使用硫醇-马来酰亚胺化学制备ADC。利用量热法、光谱法和分子模拟技术比较了ADC及其亲本IgG 1 mAb的物理稳定性。ADC的解链温度和热容都显著较低。光谱实验表明，ADC和mAb具有相似的二级和三级结构，但这些结构更容易被ADC上的热应力破坏，表明构象稳定性降低。分子模拟计算表明，在缀合后mAb的构象能显著降低。在ADC中，缀合位点周围的局部表面也变得更加疏水，这解释了ADC的较低胶体稳定性和更大的聚集倾向ADC及其亲本mAb的计算和生物物理分析提供了关于缀合对物理稳定性的影响的见解，并指出了ADC的较低结构稳定性和增加的聚集倾向背后的原因。该知识可用于设计适当的制剂以稳定ADC。
To provide a systematic biophysical approach towards a better understanding of impact of conjugation chemistry on higher order structure and physical stability of an antibody drug conjugate (ADC).ADC was prepared using thiol-maleimide chemistry. Physical stabilities of ADC and its parent IgG1 mAb were compared using calorimetric, spectroscopic and molecular modeling techniques.ADC and mAb respond differently to thermal stress. Both the melting temperatures and heat capacities are substantially lower for the ADC. Spectroscopic experiments show that ADC and mAb have similar secondary and tertiary structures, but these are more easily destabilized by thermal stress on the ADC indicating reduced conformational stability. Molecular modeling calculations suggest a substantial decrease in the conformational energy of the mAb upon conjugation. The local surface around the conjugation sites also becomes more hydrophobic in the ADC, explaining the lower colloidal stability and greater tendency of the ADC to aggregate.Computational and biophysical analyses of an ADC and its parent mAb have provided insights into impact of conjugation on physical stability and pinpointed reasons behind lower structural stability and increased aggregation propensity of the ADC. This knowledge can be used to design appropriate formulations to stabilize the ADC.