Separation of N-Glycans with Different Sialic Acid Linkages by Using Novel Superficially Porous Particle HILIC Column.
Year: 2014
Type: article
Abstract: The study of N-linked glycans is among the most challenging analytical tasks due to their complexity and variety. This difficulty is further compounded by the fact that glycans of the same composition are typically found as mixtures of multiple glycoforms differing only in branching and linkages, which can have profound biological implications. For example, previous studies have shown that the ratio of a2-3 and a2-6 linked sialic acid plays an important role in cancer biology. Consequently, differential and quantitative detection of those isomeric structures could serve as a valuable early diagnostics in oncology, which could not effectively accomplished by traditional glycomic profiling yet. Here, we studied the ability of a novel superficially porous particle (Fused-core) Penta-HILIC column to chromatographically resolve isomeric N-glycans. We found that these columns are capable of resolving N-glycans with different sialic acid linkages, as demonstrated by isolating tentative glycan isomers from model glycoproteins such as Fetuin. Structural characterization was conducted with exoglycosidase digestions and LC-MS analysis. The separation achieved by utilizing the Penta-HILIC columns on N-glycan isomers with a2-3 and a2-6 sialic acid linkages can ultimately facilitate individual glycoform identification and quantitation.
Source: Europe PMC (PubMed Central)
Authors: Yining Huang, Barry E. Boyes, Ron Orlando, Shujuan Tao
Institution: University of Georgia
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W963091820