Analysis of Glycans on Serum Proteins Using Antibody Microarrays
Antibody arrays can be employed for the profiling glycan structures on proteins. Antibody arrays capture multiple, specific proteins directly from biological samples (such as serum), and lectin and glycan-binding antibodies probe the levels of specific glycans on the captured proteins. We use a practical method of partitioning microscope slides to enable the convenient processing of many detection reagents or samples. A critical first step in the procedure is the chemical derivatization of the glycans on the spotted capture antibodies, which prevents lectin binding to those glycans. We describe those methods along with the methods for preparing and treating serum samples, running the experiments, and designing and interpreting the experiments.