Analysis of Serum Protein Glycosylation with Antibody–Lectin Microarray for High-Throughput Biomarker Screening

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 723)

Abstract

The complexity of carbohydrate structures and their derivatives makes the study of the glycome a challenging subset of proteomic research. The microarray platform has become an essential tool to characterize glycan structure and to study glycosylation-related biological interactions, by using probes as a means to interrogate the spotted or captured glycosylated molecules on the arrays. The high-throughput and reproducible nature of microarray platforms have been highlighted by their extensive applications in the field of biomarker validation, where a large number of samples must be analyzed multiple times. This chapter presents an antibody–lectin microarray approach, which allows the efficient, multiplexed study of the glycosylation of multiple individual proteins from complex mixtures with both fluorescence labeling detection and label-free detection based on mass spectrometry.

Key words

Microarray Antibody Glycoprotein Biomarker Serum Lectin MALDI Mass spectrometry 

Notes

Acknowledgements

Our work on microarray development described herein has been supported in part under grants from the National Cancer Institute under grant NCI R21 12441, R01 CA106402. This work has also received partial support from the National Institutes of Health under R01GM49500.

We would like to thank Dr. Brian Haab and Dr. Chen Songming of the Van Andel Institute for sharing with us the procedures of preparing the antibody arrays. We would also like to thank Stephanie Laurinec, Jes Pedroza, and Missy Tuck for collection of the samples used in this work.

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  1. 1.Department of ChemistryThe University of MichiganAnn ArborUSA
  2. 2.Department of Chemistry, Comprehensive Cancer CenterThe University of MichiganAnn ArborUSA
  3. 3.Department of SurgeryThe University of Michigan Medical CenterAnn ArborUSA

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