Albumin-neprilysin fusion protein: understanding stability using small angle X-ray scattering and molecular dynamic simulations
Albumin-neprilysin fusion protein: understanding stability using small angle X-ray scattering and molecular dynamic simulations
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DOI：
10.1038/s41598-020-67002-9
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发表时间：
2020-06-22
影响因子：
4.6
通讯作者：
Harris, Pernille
中科院分区：
文献类型：
--
作者：
Kulakova, Alina;Indrakumar, Sowmya;Harris, Pernille
融合技术广泛应用于蛋白质药物开发，以提高蛋白质治疗药物的活性、稳定性和生物利用度。与其他类型的生物制药一样，融合蛋白在生产和储存过程中需要保持稳定。由于高复杂性和额外的分子内相互作用，不可能根据单个蛋白质的行为来预测融合蛋白的行为。因此，了解融合蛋白在分子水平上的稳定性对生物制药的发展至关重要。目前对白蛋白-nep融合蛋白（HSA-NEP）的研究使用热化学展开结合小角度x射线散射和分子动力学模拟来显示稳定性降低和排斥性相互作用增加之间的相关性，这在大多数生物制药中是不寻常的。还可以看出，HSA-NEP不是完全灵活的：它存在于紧凑和扩展的构象中。此外，各构象的体积分数随pH的变化而变化。最后，NaCl和精氨酸的存在增加了pH 6.5时的稳定性，但降低了pH 5.0时的稳定性。
Fusion technology is widely used in protein-drug development to increase activity, stability, and bioavailability of protein therapeutics. Fusion proteins, like any other type of biopharmaceuticals, need to remain stable during production and storage. Due to the high complexity and additional intramolecular interactions, it is not possible to predict the behavior of fusion proteins based on the behavior the individual proteins. Therefore, understanding the stability of fusion proteins on the molecular level is crucial for the development of biopharmaceuticals. The current study on the albumin-neprilysin (HSA-NEP) fusion protein uses a combination of thermal and chemical unfolding with small angle X-ray scattering and molecular dynamics simulations to show a correlation between decreasing stability and increasing repulsive interactions, which is unusual for most biopharmaceuticals. It is also seen that HSA-NEP is not fully flexible: it is present in both compact and extended conformations. Additionally, the volume fraction of each conformation changes with pH. Finally, the presence of NaCl and arginine increases stability at pH 6.5, but decreases stability at pH 5.0.