N-linked glycosylation is a post-translational modification that occurs on many proteins during biosynthesis. The profile of different glycans on the protein is a critical quality attribute of some recombinant biopharmaceutical proteins including monoclonal antibodies (mAbs). Methods for profiling glycan should be robust, fast, and sensitive. Isolating glycans from proteins and tagging a label on glycans is the most commonly used technique for glycan profiling. Currently, existing protocols for sample preparation can be complicated, time-consuming, and expensive, which can limit the wide adaptation of glycan profiling methods. As a further barrier to use, an expensive ultra-high-pressure liquid chromatography (UHPLC) system is frequently required for the profile. In this article, a low cost and easily-used workflow of sample preparation is coupled with a standard high-performance liquid chromatography (HPLC) system to achieve comparable results to UHPLC. The number of steps required in the protocol and the time, as well as the cost associated with the sample preparation, is significantly reduced, while maintaining robust analytical performance. We describe the creation and validation of a human serum IgG glycan library to be used as the calibration standard, and successful profiling of glycoforms from a variety of mAbs.

N-联糖基化是翻译后修饰，在生物合成过程中发生在许多蛋白质上。蛋白上不同聚糖的概况是某些重组生物制药蛋白（包括单克隆抗体（mAb））的关键质量属性。分析聚糖的方法应坚固，快速且敏感。从蛋白质中分离聚糖并在聚糖上标记标签是最常用的聚糖谱分析技术。当前，用于样品制备的现有方案可能是复杂，费时和昂贵的，这可能限制了聚糖谱分析方法的广泛适应性。作为进一步的使用障碍，通常需要使用昂贵的超高压液相色谱（UHPLC）系统进行分析。在这篇文章中，低成本且易于使用的样品前处理流程与标准的高性能液相色谱（HPLC）系统结合使用，可获得与UHPLC相当的结果。协议中所需的步骤数量和时间以及与样品制备相关的成本已大大减少，同时保持了稳定的分析性能。我们描述了人类血清IgG聚糖文库的创建和验证，该文库将用作校准标准，并成功地从多种mAb中分析了糖型。