Particle size is a critical parameter of chromatographic resins that significantly affects protein separation. In this study, effects of resin particle sizes (31.26 μm, 59.85 μm and 85.22 μm named Aga-31, Aga-60 and Aga-85, respectively) on antibody adsorption capacity and separation performance of a hybrid biomimetic ligand were evaluated. Their performance was investigated through static adsorption and breakthrough assays to quantify static and dynamic binding capacity ( and DBC). The static adsorption results revealed that the for hIgG was 152 mg/g resin with Aga-31, 151 mg/g resin with Aga-60, and 125 mg/g resin with Aga-85. Moreover, the DBC at 10% breakthrough for hIgG with a residence time of 2 min was determined to be 49.4 mg/mL for Aga-31, 45.9 mg/mL for Aga-60, and 38.9 mg/mL for Aga-85. The resins with smaller particle sizes exhibited significantly higher capacity compared to typical commercial agarose resins and a Protein A resin (MabSelect SuRe). Furthermore, the Aga-31 resin with the hybrid biomimetic ligand demonstrated exceptional performance in terms of IgG purity (>98%) and recovery (>96%) after undergoing 20 separation cycles from CHO cell supernatant. These findings are helpful in further chromatographic resin design for the industrial application of antibody separation and purification.

中文翻译：

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探索树脂粒径对增强混合仿生配体的抗体结合能力的影响

粒径是色谱树脂的一个关键参数，显着影响蛋白质分离。本研究评估了树脂粒径（分别为 Aga-31、Aga-60 和 Aga-85，分别为 31.26 μm、59.85 μm 和 85.22 μm）对混合仿生配体的抗体吸附能力和分离性能的影响。通过静态吸附和突破分析来量化静态和动态结合能力（和 DBC），研究了它们的性能。静态吸附结果显示，Aga-31对hIgG的吸附量为152 mg/g树脂，Aga-60对hIgG的吸附量为151 mg/g树脂，Aga-85对hIgG的吸附量为125 mg/g树脂。此外，停留时间为 2 分钟的 hIgG 在 10% 突破时的 DBC 确定为：Aga-31 为 49.4 mg/mL，Aga-60 为 45.9 mg/mL，Aga-85 为 38.9 mg/mL。与典型的商业琼脂糖树脂和 Protein A 树脂 (MabSelect SuRe) 相比，具有较小粒径的树脂表现出明显更高的容量。此外，具有混合仿生配体的 Aga-31 树脂在从 CHO 细胞上清液中进行 20 个分离循环后，在 IgG 纯度 (>98%) 和回收率 (>96%) 方面表现出卓越的性能。这些发现有助于进一步设计用于抗体分离和纯化工业应用的色谱树脂。