A thermodynamic evaluation of antibody-surface interactions in multimodal cation exchange chromatography
A thermodynamic evaluation of antibody-surface interactions in multimodal cation exchange chromatography
复制标题
多模式阳离子交换色谱中抗体-表面相互作用的热力学评估
DOI：
10.1016/j.chroma.2020.461479
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发表时间：
2020
影响因子：
4.1
通讯作者：
Cramer, Steven M.
中科院分区：
文献类型：
--
作者：
Gudhka, Ronak B.;Roush, David J.;Cramer, Steven M.
在本研究中，通过保留数据的范特霍夫分析，在一系列缓冲液和盐条件下研究了两种工业单克隆抗体与多模式阳离子交换系统结合的热力学。首先在三种多峰树脂上在一系列温度和盐条件下采用等度色谱法，并在低盐和高盐条件下分析保留数据。虽然在低盐条件下，所有树脂的 mAb 保留率都会随着盐的增加而降低，但在高盐条件下，其中两种树脂的保留率会增加，这表明从静电驱动的相互作用转变为疏水性驱动的相互作用。然后使用不同温度下的保留数据来生成非线性范特霍夫图，该图适合范特霍夫方程的二次形式。在低盐条件下，两种 mAb 的保留随着温度的升高而降低，Capto MMC 和 Nuvia cPrime 上的 van't Hoff 图向下凹，而 Capto MMC ImpRes 上的 van't Hoff 图向上凹。在一些树脂上观察到关于范特霍夫图的凹度以及温度对低盐状态下有利的焓的影响的不同趋势。有趣的是，虽然在低盐条件下，Capto MMC 和 Nuvia cPrime 观察到有利的焓随着温度的变化而增加，但对于 Capto MMC ImpRes，有利的焓随着温度的升高而降低。在高盐条件下，两种 mAb 在两种 Capto 树脂上的结合始终是熵驱动的，与去溶剂化一致。虽然低盐下的负热容数据表明极性/带电基团的去溶剂化在 Capto MMC 和 Nuvia cPrime 中很重要，但正数据表明非极性基团的去溶剂化对于 Capto MMC ImpRes 更为重要。最后，高盐下的数据表明，在这些条件下，非极性基团的去溶剂化是两种 mAb 与 Capto 树脂结合的主要驱动力。
In this study, the thermodynamics of binding of two industrial mAbs to multimodal cation exchange systems was investigated over a range of buffer and salt conditions via a van't Hoff analysis of retention data. Isocratic chromatography was first employed over a range of temperature and salt conditions on three multimodal resins and the retention data were analyzed in both the low and high salt regimes. While mAb retention decreased with salt for all resins at low salts, retention increased at high salts for two of the resins, suggesting a shift from electrostatic to more hydrophobic driven interactions. The retention data at various temperatures were then employed to generate non-linear van't Hoff plots which were fit to the quadratic form of the van't Hoff equation. At low salts, retention of both mAbs decreased with increasing temperature and the van't Hoff plots were concave downward on Capto MMC and Nuvia cPrime, while being concave upward on Capto MMC ImpRes. Different trends were observed on some of the resins with respect to both the concavity of the van't Hoff plots as well as the impact of temperature on the favorable enthalpies in the low salt regime. Interestingly, while increasingly favorable enthalpy with temperature was observed with Capto MMC and Nuvia cPrime at low salt, favorable enthalpy decreased with temperature for Capto MMC ImpRes. At high salts, binding of both mAbs on the two Capto resins were consistently entropically driven, consistent with desolvation. While the negative heat capacity data at low salts indicated that desolvation of polar/charged groups were important in Capto MMC and Nuvia cPrime, the positive data suggested that desolvation of non-polar groups were more important with Capto MMC ImpRes. Finally, the data at high salts indicated that desolvation of non-polar groups was the major driver for binding of both mAbs to the Capto resins under these conditions.