Protein glycosylation profiling with PATH ® microarray slides

IN PATH non-porous nitrocellulose

Protein glycosylation analysis presents some inherent challenges due to the complexity of glycans. Determination of the mass, linkage, Isomerization and anomerization state of a protein require a combination of approaches for which relatively large amounts of proteins are needed. Such amounts are not always available when analyzing precious clinical specimens, making it difficult to extend the use of these methods for glycosylation profiling of pathological conditions.

On-chip glycan modification and probing (on-chip gmap) is a new method developed to study protein glycosylation using small amounts of protein samples. Purified proteins or tissue lysates are immobilized on a substrate and subjected to repeated cycles of glycosylation using exoglicosidases. The resulting glycans are probed with lectins. In this study PATH® Protein Microarray Slides were utilized to capture purified proteins for which information about their glycosylation state was available. Data obtained with this method were compared with MS data. The results indicate the gmap is a robust method to obtain information about protein glycosylation and can effectively complement and validate MS analysis-derived data. Learn more here: