Automated, High-Throughput IgG-Antibody Glycoprofiling Platform

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• 作者：Henning St枚ckmann ; Barbara Adamczyk ; Jerrard Hayes ; Pauline M. Rudd
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：September 17, 2013
• 年：2013
• 卷：85
• 期：18
• 页码：8841-8849
• 全文大小：529K
• 年卷期：v.85,no.18(September 17, 2013)
• ISSN：1520-6882

文摘

One of today鈥檚 key challenges is the ability to decode the functions of complex carbohydrates in various biological contexts. To generate high-quality glycomics data in a high-throughput fashion, we developed a robotized and low-cost N-glycan analysis platform for glycoprofiling of immunoglobulin G antibodies (IgG), which are central players of the immune system and of vital importance in the biopharmaceutical industry. The key features include (a) rapid IgG affinity purification and sample concentration, (b) protein denaturation and glycan release on a multiwell filtration device, (c) glycan purification on solid-supported hydrazide, and (d) glycan quantification by ultra performance liquid chromatography. The sample preparation workflow was automated using a robotic liquid-handling workstation, allowing the preparation of 96 samples (or multiples thereof) in 22 h with excellent reproducibility and, thus, should greatly facilitate biomarker discovery and glycosylation monitoring of therapeutic IgGs.