Molecular computations of preferential interactions of proline, arginine.HCl, and NaCl with IgG1 antibodies and their impact on aggregation and viscosity.
Molecular computations of preferential interactions of proline, arginine.HCl, and NaCl with IgG1 antibodies and their impact on aggregation and viscosity.
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DOI：
10.1080/19420862.2020.1816312
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发表时间：
2020-01
期刊：
影响因子：
5.3
通讯作者：
Trout BL
中科院分区：
文献类型：
--
作者：
Cloutier TK;Sudrik C;Mody N;Hasige SA;Trout BL
辅料与抗体表面的优先相互作用决定了它们对抗体在溶液中的稳定性的影响。我们通过实验和模拟探讨了三种治疗相关的IgG1抗体与脯氨酸、精氨酸、盐酸盐和氯化钠的优先相互作用。通过模拟，我们研究了赋形剂如何与可变区(Fv)中不同类型的表面补丁相互作用。例如，Pro与芳香族表面的相互作用最强，Arg.HCl位于负残基附近，而负残基和某些疏水区域排除了氯化钠。不同辅料在同一抗体表面上相互作用的差异可能是抗体在每种辅料中聚集、粘度和自结合行为不同的原因。Pro减少了所有三种抗体的自结合，并通过缔合限制聚集机制减少了抗体的聚集。Arg.HCl和氯化钠对聚集性和粘度的影响高度依赖于表面电荷分布和对高度疏水斑块的排斥程度。在pH为5.5时，两者都倾向于增加Fv上带强正电荷的抗体的聚集，而只有氯化钠减少了Fv上带大负电荷的抗体的聚集。Arg.HCl通过疏水性驱动机制或电荷驱动机制降低抗体的粘度。对这些数据的分析为理解氨基酸和离子辅料如何与不同的蛋白质表面相互作用，以及这些相互作用如何转化为观察到的稳定性行为提供了一个框架。
Preferential interactions of excipients with the antibody surface govern their effect on the stability of antibodies in solution. We probed the preferential interactions of proline, arginine.HCl (Arg.HCl), and NaCl with three therapeutically relevant IgG1 antibodies via experiment and simulation. With simulations, we examined how excipients interacted with different types of surface patches in the variable region (Fv). For example, proline interacted most strongly with aromatic surfaces, Arg.HCl was included near negative residues, and NaCl was excluded from negative residues and certain hydrophobic regions. The differences in interaction of different excipients with the same surface patch on an antibody may be responsible for variations in the antibody's aggregation, viscosity, and self-association behaviors in each excipient. Proline reduced self-association for all three antibodies and reduced aggregation for the antibody with an association-limited aggregation mechanism. The effects of Arg.HCl and NaCl on aggregation and viscosity were highly dependent on the surface charge distribution and the extent of exclusion from highly hydrophobic patches. At pH 5.5, both tended to increase the aggregation of an antibody with a strongly positive charge on the Fv, while only NaCl reduced the aggregation of the antibody with a large negative charge patch on the Fv. Arg.HCl reduced the viscosities of antibodies with either a hydrophobicity-driven mechanism or a charge-driven mechanism. Analysis of this data presents a framework for understanding how amino acid and ionic excipients interact with different protein surfaces, and how these interactions translate to the observed stability behavior.