Glycosylation Profiling of IL-23: Determination of N-Glycosylation Sites and Structure Characterization of the Oligosaccharides by Mass Spectrometry

摘要: As a new member of the growing family of interleukin (IL)-12-related cytokines, IL-23 is a key immunoregulatory cytokine that coordinates innate and adaptive immune responses. IL-23 promotes end-stage inflammation, which makes it and the proteins in its down-stream pathway potential targets for the treatment of immune-mediated diseases. As a heterodimeric cytokine that comprises of disulfide-linked p19 and p40 subunits, IL-23 has four potential N-glycosylation sites 125NKTF, 135NYSG, 222NYTS, and 3 deg 3NASI. Structural characterization of the glycoforms of IL-23 has not yet been reported. In this presentation, we describe the use of mass spectrometry (MS) to establish the glycosylation profiles of recombinant IL-23, which includes defining glycosylation sites, the degree of occupancy of each glycosylation site, and the structures of the oligosaccharides attached to each site.