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固定化的酶级联用于有针对性的糖基化

Elli Makrydaki¹, Roberto Donini², Anja Krueger²

¹Department of Chemical Engineering, Imperial College London, London, UK.

Nature chemical biology

|February 6, 2024

概括

研究人员开发了一种用于受控蛋白质糖化酶的新平台,创造了定制的糖结构. 这种方法产生了近同质的糖形式,以改善蛋白质功能和临床评估.

科学领域:

生物化学和分子生物学
葡萄糖生物学葡萄糖生物学
蛋白质工程是指蛋白质工程.

背景情况:

糖基化是一种关键的翻译后修改,影响蛋白质折叠,稳定性和功能.
甘氨基化固有的异质性和非模板性质源于酶乱交.
实现受控和均的糖化是治疗性蛋白质开发和功能研究的关键.

研究的目的:

引入一个新的平台,Sugar-TARGET,用于量身定制和受控的N结合甘化在体外.
为了证明该平台能够从各种蛋白质来源生成近同质的甘油形式的能力.
评估对治疗性蛋白质的特定糖基化配置的增强,例如银河基化.

主要方法:

开发一种单步固定/净化方法,用于酶生产.
使用固定酶,模仿自然甘氨基化途径的序列反应级联的构建.
固定β-1,4-银酸转移酶的应用,以修改免疫球蛋白G (IgG) 糖形.

主要成果:

糖-TARGET平台可为定制糖结构提供可控的,顺序的糖化反应.
从各种蛋白质中成功地生成了近同质的葡萄糖形式,使它们的葡萄糖化简介变得人性化.
对三种IgG的固定β-1,4-银酸转移酶治疗导致80.2-96.3%的终端银酸转化.
证明了酶可循环,可延长反应时间 (>80小时).

结论:

糖目标平台提供了一个模块化,可重复使用和易于实施的解决方案,用于生产均的糖结构.
这项技术有助于生成量身定制的葡萄糖蛋白,以提高功能和临床评估.
在体外可以实现受控的糖化,克服自然异质过程的局限性.

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