Pngase f glycosidase

High purity LC-MS grade water and acetonitrile containing formic acid used for mobile phase preparation, and ammonium bicarbonate reagent were purchased from J.T. Baker (Phillipsburg, NJ). RapiGest™ surfactant and [Glu1]-Fibrinopeptide B lock mass standard (GluFib) were purchased from Waters (Milford, MA). Formic acid, urea, and Zeba™ spin desalting column (7K MWCO) were purchased from Pierce (Rockford, IL). Dithiothreitol (DTT) and iodoacetamide were purchased from ThermoFisher Life Technologies (Grand Island, NY) and Sigma-Aldrich (St. Louis, MO), respectively. Trypsin (modified sequencing grade), chymotrypsin, IdeS (immunoglobulin G-degrading enzyme of Streptococcus pyogenes), and PNGase F glycosidase were purchased from Promega (Madison, WI).

HA-SS-np materials were produced at the Vaccine Production Program Laboratory (Gaithersburg, MD). HA-SS-np formulated in one of the five formulation buffers (containing sodium phosphate, sodium chloride, and sucrose, pH 7.2) was used for investigational purposes, and a HA-SS-np interim reference material (in 1x PBS) was used as a control. Reagents included LC-MS grade 0.1% formic acid in water (mobile phase A), 0.1% formic acid acetonitrile (mobile phase B), and ammonium bicarbonate that were purchased from JT Baker (Phillipsburg, NJ). RapiGest^™ surfactant and [Glu1]-Fibrinopeptide B lock mass standard were purchased from Waters (Milford, MA). Formic acid, urea, and Zeba^™ desalting columns (7K MWCO) were purchased from Pierce (Rockford, IL). Dithiothreitol (DTT) and iodoacetamide (IAM) were purchased from ThermoFisher Life Technologies (Grand Island, NY) and Sigma Aldrich (St. Louis, MO), respectively. Chymotrypsin and PNGase F glycosidase were purchased from Promega (Madison, WI); Lys-C was purchased from New England Biolabs (Ipswich, MA). Method details of the stability-indicating analyses (rCGE and SEC) are described in the online supporting information.

High purity LC-MS grade water and acetonitrile containing formic acid used for mobile phase preparation and ammonium bicarbonate reagent were purchased from J.T. Baker (Phillipsburg, NJ). RapiGest™ surfactant and [Glu1]-Fibrino-peptide B lock mass standard (GluFib) were purchased from Waters (Milford, MA). Formic acid, urea, and Zeba™ spin desalting column (7 K MWCO) were purchased from Pierce (Rockford, IL). Dithiothreitol (DTT) and iodoacetamide were purchased from ThermoFisher Life Technologies (Grand Island, NY) and Sigma-Aldrich (St. Louis, MO), respectively. Trypsin (modified sequencing grade), chymotrypsin, and PNGase F glycosidase and endoglycosidase (Endo) H were purchased from Promega (Madison, WI). Alpha 1–2,3 mannosidase was purchased from New England Biolabs (Ipswich, MA).