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在抗体净化过程中,通过集成的特定位点固定和分子动力学来阐明M蛋白的性能及其结合模式

Alexey Tarabarov¹, Xingxing Wang¹, Yige Lu¹

¹Zhejiang Provincial Key Lab for Chemical and Biological Processing Technology of Farm Products, School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou, 310023, People's Republic of China.

International journal of biological macromolecules

|October 15, 2025

概括

磁纳米颗粒上的蛋白M-8His提供了广泛的抗体净化. 分子动力学揭示了不同的结合机制,解释了可变性能和指导优化工业抗体净化.

科学领域:

生物技术是生物技术.
蛋白质工程是指蛋白质工程.
纳米技术纳米技术

背景情况:

使用蛋白A/G/L的传统抗体净化方法由于特定的结合区域而存在局限性.
蛋白M是一种工程变异 (蛋白M-8His),通过可变光链域显示宽谱抗体结合.
蛋白M-8His在不同抗体中的可变性能背后的分子机制尚不清楚.

研究的目的:

研究蛋白M-8His用于抗体净化的分子机制.
在纳米颗粒上开发一种使用固定蛋白M-8His的抗体净化平台.
解释蛋白M-8His在不同抗体中的可变结合性能.

主要方法:

在Ni2+修饰的超偏磁纳米粒子 (NiNPs) 上对M-8His蛋白的特定位点固定.
纳米粒子结合能力和抗体活性保留的表征.
分子动力学 (MD) 模拟和能量分解分析以研究结合相互作用.

主要成果:

开发的平台 (MNiNPs) 显示高蛋白M-8His结合能力 (74.41毫克/克) 和宽谱抗体结合 (达达52.01毫克/克的trastuzumab scFv).
储存15天后,抗体活性保持在76.1%.
MD模拟显示了不同的相互作用模式:对trastuzumab scFv的依赖histidine的化和对S2P6和76E1抗体的混合相互作用,解释了性能变化.
分析显示了差异性的结合域:trastuzumab scFv和76E1 Fab的Fv域,以及S2P6 Fab的常数域.

结论:

提供了对蛋白质M-8及其与抗体的相互作用的机制性见解.
不同的结合模式和域相互作用解释了蛋白M-8His.的可变性能.
这些发现支持优化M-8His蛋白用于工业抗体净化应用.

关键词:

抗体净化抗体净化分子动力学分子动力学蛋白质M是一种蛋白质M.网站特定的蛋白质固定化超偏磁纳米粒子超偏磁纳米粒子

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