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用[Bathophenathroline:metal]复合体对Fc融合蛋白进行净化

Thisara Jayawickrama Withanage¹, Ron Alcalay², Olga Krichevsky¹

¹Department of Chemical Sciences, Ariel University, Ariel 4070000, Israel.

Antibodies (Basel, Switzerland)

|February 21, 2025

概括

这项研究引入了一种使用芳香复合物和PEG-6000的新型Fc-fusion蛋白质净化方法,实现高产量并保持酶活性. 该过程是可扩展的,并且对工业应用具有成本效益.

科学领域:

生物化学生物化学
蛋白质净化蛋白质净化
生物技术是生物技术.

背景情况:

传统的Fc融合蛋白质净化依赖于染色学,这可能是昂贵和复杂的.
开发替代的,具有成本效益的净化策略对于工业生物制造至关重要.

研究的目的:

评估使用沉芳香复合物的替代Fc融合蛋白净化方法.
评估这种新型净化技术的效率,产量和可扩展性.

主要方法:

使用巴托芬罗林 - 金属复合物 (Zn2+或Cu2+) 和PEG-6000来净化与人类IgG1 Fc域 (AChE-Fc) 融合的复合人性乙胆酶.
一个三步过程,包括捕获,用NaCl去除杂质,用酸盐缓冲剂提取.
使用动态光散射 (DLS) 和原生PAGE进行蛋白质聚合的分析.

主要成果:

[(batho) 3:Zn2+]复合物成功净化了ACHE-Fc,保持了充分的酶活性.
在有效的化剂循环利用 (95-97%) 的情况下,实现了大约83-88%的总蛋白质产量.
证明了成功的30倍工艺升级,试剂的比例增加.

结论:

这种新的净化方法为Fc融合蛋白质的传统染色学提供了经济和实用的替代方案.
芳香复合物的使用提供了一个无联体和无介质的净化策略.
该过程具有可扩展性,并且在生物制药制造中显示出工业应用的潜力.

关键词:

在Fc-融合蛋白质中.乙烯基胆化酶的使用巴托芬罗林:Zn2+ 复合物非染色学净化净化方法

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