Figure 1. Workflow for the discovery of human and porcine glycans.
A. This study compares the N-linked glycomes of humans, domestic pigs, GGTA1 knockout pigs and GGTA1/CMAH knockout pigs. B. Serum samples were treated with the deglycosylating enzyme, PNGaseF, to remove N-linked glycans from serum proteins. C. Native glycans were stabilized by reduction and permethylation on solid-phase sodium hydroxide spin columns. D. Collision induced dissociation of permethylated glycans resulted in predictable fragmentation that was analyzed using the nomenclature of Domon and Costello (19).