An experimental approach probing the conformational transitions and energy landscape of antibodies: a glimmer of hope for reviving lost therapeutic candidates using ionic liquid.
An experimental approach probing the conformational transitions and energy landscape of antibodies: a glimmer of hope for reviving lost therapeutic candidates using ionic liquid.
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DOI：
10.1039/d1sc02520a
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发表时间：
2021-07-14
期刊：
影响因子：
8.4
通讯作者：
Hallett JP
中科院分区：
文献类型：
--
作者：
Shmool TA;Martin LK;Bui-Le L;Moya-Ramirez I;Kotidis P;Matthews RP;Venter GA;Kontoravdi C;Polizzi KM;Hallett JP
了解蛋白质在不同环境条件下的折叠对于预测蛋白质结构和开发创新的抗体配方至关重要。虽然通过计算方法对折叠和解折叠的热力学和动力学进行了广泛的研究，但缺乏确定抗体构象转换途径的实验方法。为了填补这一空白，我们在离子液体(IL)胆碱二氢磷酸存在和不存在的情况下，用不同的辅料制备了一系列独特的配方，其中包含高浓度的嵌合免疫球蛋白G4(IgG4)抗体。我们通过变温、圆二色谱和生物层干涉分析确定了不同辅料和IL对蛋白质热稳定性和结构稳定性的影响。为了进一步合理地观察到构象随温度的变化，我们对不同配方中的单个抗体结合片段在低温和高温下进行了分子动力学模拟。我们发展了一种方法来研究生物分子的构象转变和相关的热力学，并展示了IL诱导的构象转变。我们发现，构象变化倾向的增加是由磷酸二氢阴离子优先结合到抗体片段所驱动的。最后，我们发现含有IL与糖、氨基酸和表面活性剂的配方是稳定蛋白质防止构象破坏和聚集的一个有前途的候选方案。我们希望，最终，我们可以帮助我们了解抗体和蛋白质错误折叠现象的稳定性的分子基础，并提供新的候选配方，有可能重振失去的候选治疗药物。探索用于药物设计的生物分子的能量格局和热力学。
Understanding protein folding in different environmental conditions is fundamentally important for predicting protein structures and developing innovative antibody formulations. While the thermodynamics and kinetics of folding and unfolding have been extensively studied by computational methods, experimental methods for determining antibody conformational transition pathways are lacking. Motivated to fill this gap, we prepared a series of unique formulations containing a high concentration of a chimeric immunoglobin G4 (IgG4) antibody with different excipients in the presence and absence of the ionic liquid (IL) choline dihydrogen phosphate. We determined the effects of different excipients and IL on protein thermal and structural stability by performing variable temperature circular dichroism and bio-layer interferometry analyses. To further rationalise the observations of conformational changes with temperature, we carried out molecular dynamics simulations on a single antibody binding fragment from IgG4 in the different formulations, at low and high temperatures. We developed a methodology to study the conformational transitions and associated thermodynamics of biomolecules, and we showed IL-induced conformational transitions. We showed that the increased propensity for conformational change was driven by preferential binding of the dihydrogen phosphate anion to the antibody fragment. Finally, we found that a formulation containing IL with sugar, amino acids and surfactant is a promising candidate for stabilising proteins against conformational destabilisation and aggregation. We hope that ultimately, we can help in the quest to understand the molecular basis of the stability of antibodies and protein misfolding phenomena and offer new candidate formulations with the potential to revive lost therapeutic candidates. Probing the energy landscape and thermodynamics of biomolecules for drug design.
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影响因子：
4.9
作者：
Cloutier, Theresa;Sudrik, Chaitanya;Trout, Bernhardt L.
通讯作者：
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影响因子：
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影响因子：
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作者：
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通讯作者：
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