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Determination of the degree of PEGylation of protein bioconjugates using data from proton nuclear magnetic resonance spectroscopy

机译:使用质子核磁共振光谱数据确定蛋白质生物缀合物的PEG化程度

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The average number of methoxy poly(ethylene glycol) (mPEG) chains grafted to a protein – also known as the degree of PEGylation – is a fundamental parameter for characterizing a bioconjugate. The degree of PEGylation is typically determined by chromatographic or electrophoretic methods, which are subject to certain biases. This contribution describes an analytical approach alongside technical precautions for quantitatively determining the degree of PEGylation of protein bioconjugates by1H NMR spectroscopy. An accompanying dataset, corresponding to the raw1H NMR spectra of thirteen bioconjugates with different degrees of PEGylation and different mPEG molecular weights, is provided for the reader to become familiar with the analysis. The exemplary bioconjugate system used in this Data article is the enzyme glutamate dehydrogenase (GDH) modified with multiple copies of mPEG (0.5–20 kDa). These bioconjugates correspond to those discussed in-depth in the article “Mechanisms of activity loss for a multi-PEGylated protein by experiment and simulation” by Zaghmi et?al., 2019 The described approach to calculate degree of PEGylation is quantitative, applicable to other proteins, and can be adapted to other types of polymers.
机译:嫁接到蛋白质上的甲氧基聚(乙二醇)(mPEG)链的平均数量(也称为PEG化程度)是表征生物缀合物的基本参数。 PEG化的程度通常通过色谱法或电泳法来确定,这受到某些偏差的影响。该贡献描述了一种分析方法以及技术预防措施,用于通过1H NMR光谱法定量确定蛋白质生物缀合物的PEG化程度。随附的数据集对应于十三种具有不同PEG化程度和不同mPEG分子量的生物缀合物的raw 1 H NMR光谱,供读者熟悉分析。本数据文章中使用的示例性生物缀合物系统是用多份mPEG(0.5-20 kDa)修饰的谷氨酸脱氢酶(GDH)。这些生物结合物对应于Zaghmi等人在2019年发表的文章“通过实验和模拟使多聚乙二醇化蛋白质的活性丧失的机理”中深入讨论的那些生物结合物。所述计算PEG化程度的方法是定量的,适用于其他蛋白质,并且可以适应其他类型的聚合物。

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