Discovery and Characterization of a Peptide Motif That Specifically Recognizes a Non-native Conformation of Human IgG Induced by Acidic pH Conditions
Discovery and Characterization of a Peptide Motif That Specifically Recognizes a Non-native Conformation of Human IgG Induced by Acidic pH Conditions
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DOI：
10.1074/jbc.m807618200
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发表时间：
2009-04-10
影响因子：
4.8
通讯作者：
Sugimura, Kazuhisa
中科院分区：
文献类型：
--
作者：
Sakamoto, Kotaro;Ito, Yuji;Sugimura, Kazuhisa
在治疗性抗体制备中，酸性pH条件通常用于从IgG的蛋白A亲和柱洗脱或用于其病毒灭活。将IgG暴露于低pH条件诱导构象变化，导致其功能损伤或丧失，尽管其机制尚未完全阐明。在这项研究中，使用随机肽T7噬菌体展示库，我们分离出一个独特的和新的肽基序，特异性地识别由低pH处理产生的人IgG的非天然构象（酸性构象），但不是天然构象。我们研究了生成条件和使用的肽基序作为亲和配体的酸构象的生化性质。酸性构象异构体在酸性pH值（25摄氏度）下很容易生成。这里分离的肽可能有助于阐明在酸暴露以及人IgG的储存期间抗体功能障碍或聚集的机制。
In therapeutic antibody preparation, acidic pH conditions are generally used for elution from Protein A affinity column of IgG or for its viral inactivation. Exposing IgG to low pH conditions induces conformational changes, leading to its functional damage or loss, although the mechanisms have not been fully elucidated. In this study using random peptide T7 phage display libraries, we isolated a unique and novel peptide motif that specifically recognized the non-native conformer ( acid conformer) of human IgG that was generated by the low pH treatment, but not the native conformer. We examined the generation conditions and biochemical properties of acid conformer using the peptide motif as an affinity ligand. The acid conformer was easily generated at acidic pH (25 degrees C). The peptides isolated here could contribute to the elucidation of the mechanisms of antibody dysfunction or aggregation during acid exposure as well as storage of human IgG.