A Simple Strategy to Eliminate Hexosylation Bias in the Relative Quantification of N-Glycosylation in Biopharmaceuticals.,Angewandte Chemie International Edition

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A Simple Strategy to Eliminate Hexosylation Bias in the Relative Quantification of N-Glycosylation in Biopharmaceuticals.
Angewandte Chemie International Edition ( IF 16.1 ) Pub Date : 2020-06-04 , DOI: 10.1002/anie.202002147
Wolfgang Esser-Skala _{1,

2} , Therese Wohlschlager _{1,

2} , Christof Regl _{1,

2} , Christian G Huber _{1,

2}

Affiliation

N‐glycosylation may affect the safety and efficacy of biopharmaceuticals and is thus monitored during manufacturing. Mass spectrometry of the intact protein is increasingly used to reveal co‐existing glycosylation variants. However, quantification of N‐glycoforms via this approach may be biased by single hexose residues as introduced by glycation or O‐glycosylation. Herein, we describe a simple strategy to reveal actual N‐glycoform abundances of therapeutic antibodies, involving experimental determination of glycation levels followed by computational elimination of the “hexosylation bias”. We show that actual N‐glycoform abundances may significantly deviate from initially determined values. Indeed, glycation may even obscure considerable differences in N‐glycosylation patterns of drug product batches. Our observations may thus have implications for biopharmaceutical quality control. Moreover, we solve an instance of the problem of isobaricity, which is fundamental to mass spectrometry.

中文翻译：

一种在生物药物中N-糖基化相对定量中消除己糖基化偏见的简单策略。

N-糖基化可能会影响生物药物的安全性和有效性，因此在生产过程中会受到监控。完整蛋白质的质谱法越来越多地用于揭示共存的糖基化变体。但是，通过这种方法对N-糖型的定量可能会因糖基化或O-糖基化引入的单个己糖残基而产生偏差。本文中，我们描述了一种简单的策略来揭示治疗性抗体的实际N-糖型丰度，包括实验确定糖基化水平，然后通过计算消除“己糖基化偏倚”。我们表明，实际的N-糖型丰度可能与最初确定的值明显不同。确实，糖基化甚至可以掩盖药品批次中N-糖基化模式的巨大差异。因此，我们的观察结果可能会对生物制药质量控制产生影响。此外，我们解决了等压问题的一个实例，这是质谱学的基础。

更新日期：2020-06-04

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