title: Quantification of each serogroup polysaccharide of Neisseria meningitidis in A/C/Y/W-135-DT conjugate vaccine by high-performance anion-exchange chromatography-pulsed amperometric detection analysis.
abstract: Invasive bacterial meningitis caused by Neisseria meningitidis can be prevented by active immunization with meningococcal polysaccharide or polysaccharide-protein conjugate vaccines. In a tetravalent A/C/Y/W-135-DT meningococcal conjugate vaccine vial, or in a final formulated bulk, accurate identification and quantification of each polysaccharide are critical in product release. Determination of sialic acid serogroups (C, W-135, and Y) unambiguously is complex since all these serogroups contribute to the sialic acid monosaccharide peaks that overlap in the high-performance anion-exchange chromatography-pulsed amperometric detection (HPAEC-PAD). We report a quantification method that involves generation of monosaccharide standard plots for respective sugars mannosamine-6-phosphate, sialic acid, galactose- and glucose-derived from hydrolysis of mixtures of the four serogroups A, C, W, and Y reference polysaccharides. These plots were then used to obtain the unknown polysaccharide concentrations of A/C/Y/W-135 in vialed vaccine or from formulated final bulks. We also present our results of the HPAEC-PAD profiles on groups C, W-135, and Y polysaccharides when hydrolyzed individually and/or in mixtures to discuss the individual sialic acid peak contributions.
authors.affiliation: JNI Medical Corporation , 2720 N 84th Street, Omaha, Nebraska 68134, United States.
authors.forename: Seshu Kumar
authors.initials: SK
authors.lastname: Gudlavalleti
authors.forename: Erika Nicole
authors.initials: EN
authors.lastname: Crawford
authors.forename: Jeffery David
authors.initials: JD
authors.lastname: Harder
authors.forename: Jeeri Raghava
authors.initials: JR
authors.lastname: Reddy
mesh_terms: D002626:Chemistry, Pharmaceutical; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D005690:Galactose / Q000737:chemistry; D005947:Glucose / Q000737:chemistry; D006868:Hydrolysis; D008358:Mannose / Q000737:chemistry; D022401:Meningococcal Vaccines / Q000737:chemistry*; D038381:Neisseria meningitidis, Serogroup A / Q000737:chemistry*; D038542:Neisseria meningitidis, Serogroup C / Q000737:chemistry*; D038582:Neisseria meningitidis, Serogroup W-135 / Q000737:chemistry*; D038581:Neisseria meningitidis, Serogroup Y / Q000737:chemistry*; D011134:Polysaccharides / Q000032:analysis* / Q000276:immunology; D012794:Sialic Acids / Q000737:chemistry; D018074:Vaccines, Conjugate / Q000032:analysis* / Q000276:immunology
chemical_list: D022401:Meningococcal Vaccines; D011134:Polysaccharides; D012794:Sialic Acids; D018074:Vaccines, Conjugate; D005947:Glucose; D008358:Mannose; D005690:Galactose