title: Size exclusion chromatography of polysaccharides with reverse phase liquid chromatography.
abstract: This work describes the use of a reverse phase (RP) column for size-based separation of hydrophilic polysaccharides. The entire separation window (before and after void volume) is used to provide unique separation of polysaccharides from other components in a glycol-conjugate vaccine or complex media of fermentation broth. The technique has also been applied to the separation of polysaccharides of different sizes. The effect of chromatographic parameters including type of packing material (CN, C8 and C18), pore size (80 and 300Å) of stationary phase, concentration of organic solvent on the separation was investigated. In addition, characterization of size-based separation was evaluated by coupling of RP column with multi-angle laser light scattering (MALLs) detector.
authors.affiliation: Analytical R&D, Pfizer BioTherapeutics R&D Pharmaceutical Sciences, 700 Chesterfield Parkway West, Chesterfield, MO 63017, United States. Electronic address: Yan.He2@pfizer.com.
authors.forename: Yan
authors.initials: Y
authors.lastname: He
authors.affiliation: Analytical R&D, Pfizer BioTherapeutics R&D Pharmaceutical Sciences, 700 Chesterfield Parkway West, Chesterfield, MO 63017, United States.
authors.forename: Weiying
authors.initials: W
authors.lastname: Hou
authors.affiliation: Analytical R&D, Pfizer BioTherapeutics R&D Pharmaceutical Sciences, 700 Chesterfield Parkway West, Chesterfield, MO 63017, United States.
authors.forename: Melissa
authors.initials: M
authors.lastname: Thompson
authors.affiliation: Analytical R&D, Pfizer BioTherapeutics R&D Pharmaceutical Sciences, 700 Chesterfield Parkway West, Chesterfield, MO 63017, United States.
authors.forename: Heidi
authors.initials: H
authors.lastname: Holovics
authors.affiliation: Analytical R&D, Pfizer BioTherapeutics R&D Pharmaceutical Sciences, 700 Chesterfield Parkway West, Chesterfield, MO 63017, United States.
authors.forename: Tracy
authors.initials: T
authors.lastname: Hobson
authors.affiliation: Analytical R&D, Pfizer BioTherapeutics R&D Pharmaceutical Sciences, 700 Chesterfield Parkway West, Chesterfield, MO 63017, United States.
authors.forename: Michael T
authors.initials: MT
authors.lastname: Jones
mesh_terms: D002850:Chromatography, Gel; D056148:Chromatography, Reverse-Phase / Q000295:instrumentation; D045424:Complex Mixtures / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D057927:Hydrophobic and Hydrophilic Interactions; D008970:Molecular Weight; D010316:Particle Size; D011134:Polysaccharides / Q000737:chemistry / Q000302:isolation & purification*; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D045424:Complex Mixtures; D011134:Polysaccharides; D011506:Proteins; D004247:DNA
keywords: Polysaccharide; Reverse phase liquid chromatography; Size exclusion chromatography; Void volume