Journal: BioMed Research International

Article Title: Development of Two Analytical Methods Based on Reverse Phase Chromatographic and SDS-PAGE Gel for Assessment of Deglycosylation Yield in N-Glycan Mapping

doi: 10.1155/2018/3909674

Figure Lengend Snippet: Chromatogram of dextran ladder (a), N -glycan profile of NIST mAb prepared with GlycanAssure HyPerformance APTS kit and analyzed on Accucore 150 Amide HILIC LC Column and Vanquish Horizon UHPLC equipped with FLD zoomed out (b) and zoomed in views (c).

Article Snippet: Materials and Reagents SimplyBlue™ SafeStain from Novex by Life Technologies (Cat# LC6065), NuPAGE MOPS SDS running buffer (P/N# NP0001), NuPAGE LDS sample buffer (4x) (Cat# NP0008), NuPAGE Bis-Tris Mini Gels (Cat# IM-8042), GlycanAssure HyPerformance APTS kit (Cat# A33953), 1 M DTT (P/N# P2325), formic acid, 99.5+%, Optima™ LC/MS grade (P/N#A117-50) from Thermo Fisher Scientific, HPLC grade 100% acetonitrile (P/N#AC610010040) from Fisher Scientific, Aeris™ 3.6 μ m WIDEPORE XB-C18 (200 Å, LC Column 150 × 4.6 mm) (P/N 00F-4482-E0) from Phenomenex, Thermo Scientific™ Accucore™ 150 Amide HILIC LC Column (2.6 μ m, 150 A, 2.1 × 150 mm), and Thermo Scientific Vanquish™ Horizon UHPLC System, glycoproteins of RNase B and bovine fetuin from Sigma (P/N#J62334, lot R112C021) and QA-Bio (P/N# GCP 500 μ g, lot# B1B2-04), IgG1κ monoclonal antibody (NIST mAb #8671 (lot# 14HB-D-001)) in 12.5 mmol/L L-histidine, 12.5 mmol/L L-histidine HCl (pH 6.0) from NIST were used.

Techniques: Hydrophilic Interaction Liquid Chromatography