Characterization of IgG N-Glycans Employing a Microfluidic Chip that Integrates Glycan Cleavage, Sample Purification, LC Separation, and MS Detection

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• 作者：Maggie A. Bynum ; Hongfeng Yin ; Katherine Felts ; Yvonne M. Lee ; Craig R. Monell ; Kevin Killeen
• 刊名：Analytical Chemistry
• 出版年：2009
• 出版时间：November 1, 2009
• 年：2009
• 卷：81
• 期：21
• 页码：8818-8825
• 全文大小：342K
• 年卷期：v.81,no.21(November 1, 2009)
• ISSN：1520-6882

文摘

A novel polymeric microfluidic device with an on-chip enzyme reactor has been developed for the characterization of recombinant glycoproteins. The enzyme reactor chip packed with PNGase F-modified solid support material was combined with a microfluidic glycan cleanup chip and a commercially available HPLC-chip to perform glycoprotein deglycosylation, protein removal, glycan capture, glycan LC separation, and nanoelectrospray into a time-of-flight mass spectrometry (TOF-MS) system. With this integrated chip, the combined sample preparation and sample analysis time was reduced from multiple hours to less than 10 min. A once tedious and time-consuming glycan analysis workflow is now integrated into an HPLC-chip device. Glycan profiling analysis has been achieved with as little as 100 ng of monoclonal antibody. Furthermore, a single chip was shown to retain activity and perform equivalently for over 250 replicate glycan profiles from a recombinant antibody.