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Optimization of GlycoWorks HILIC SPE for the Quantitative and Robust Recovery of N-Linked Glycans from mAb-Type Samples

11 Aug 2013

This application note evaluates HILIC SPE sample preparation to ensure quantitative recovery of both unlabeled and labeled N-glycans. A test mixture, including a complex array of 2-AB labeled human IgG glycans spiked with both high mannose and trisialylated glycans, was used to interrogate and optimize SPE recoveries as well as study the robustness of optimized elution conditions.

ACQUITY UPLC H-Class PLUS Bio System

Waters

The ACQUITY UPLC H-Class PLUS Bio System delivers the benefits of UPLC's resolution, sensitivity, and throughput in a system purpose-built for the analysis of proteins, peptides, nucleic acids, and glycans.

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GlycoWorks HILIC uElution Plate

Waters

The GlycoWorks Product Line was developed and designed to compliment the existing Waters Glycan Application Solution. Concentrating on 2-AB labeled N-Glycan, this sample preparation offering and complimentary standards provides a unique, comprehensive yet versatile solution. The GlycoWorks HILIC uElution Plate is ideal for removing contaminants like salts and detergents from hydrophilic analytes, such as carbohydrates, prior to mass spectrometry analysis.It enables elution volumes as low as 25 µL, thereby eliminating time-consuming evaporation steps. The eluted carbohydrates can be analyzed directly by MALDI MS without any prior derivatization.This plate can be used alone for the purpose of extracting carbohydrates, or it can be used as part of our glycan analysis kit (Waters MassPREP Glycoanalysis Kit) to clean up the glycans released from glycoproteins after deglycosylation.

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Optimization of GlycoWorks HILIC SPE for the Quantitative and Robust Recovery of N-Linked Glycans from mAb-Type Samples