LCMS data processing for Sulf2 digesiton

Partialy digestion of porcine intestinal mucosa heparan sulfate was fractionated by off-line SEC column. The fraction corresponding to DP4 (DP: degree of polymerization), DP6 and DP8 oligosaccharides were collected for HSulf2 treatment and HILIC LC-MS analysis on an Agilent 6520 QTOF MS with chip cube interface.

The HILIC LC-MS spectra were decovoluted using the combination of customized Decon2Ls/DeconTools Autoprocessor from Pacific Northwest National Laboratory and Progenesis (Nonlinear Dynamic Limited). The profiling of the HS compositions was achieved using in-house fitting programs. The composition coding of the HS and heparin is presented as [ΔHexA, HexA, GlcN, Ac, SO3] (ΔHexA: ∆4,5-unsaturated hexuronic acid; HexA: hexuronic acid; GlcN: glucosamine; Ac: acetyl group; SO3: sulfate).