title: Quantitation of low level unconjugated polysaccharide in tetanus toxoid-conjugate vaccine by HPAEC/PAD following rapid separation by deoxycholate/HCl.
abstract: A simple and rapid acid precipitation method has been applied successfully for separating free capsular polysaccharide of Haemophilus influenzae type b (polyribosyl ribitol phosphate, PRP) from PRP tetanus toxoid conjugate (PRP-T) in a final dosage amount of low-level materials. The unconjugated PRP was found to stay in the supernatant without precipitation, while conjugated PRP-T was fully precipitated. High performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) has been applied for analysis of the PRP content in the supernatant after the separation. This method requires minimum sample handling and is specific, sensitive and reproducible making it suitable for release and stability testing of PRP-T in final containers.
authors.affiliation: Research Test Development and Clinical Serology, Pasteur Merieux Connaught, Swiftwater, PA 18370-0187, USA. plei@us.pmc-vacc.com
authors.forename: Q P
authors.initials: QP
authors.lastname: Lei
authors.forename: D H
authors.initials: DH
authors.lastname: Lamb
authors.forename: R
authors.initials: R
authors.lastname: Heller
authors.forename: P
authors.initials: P
authors.lastname: Pietrobon
mesh_terms: D000837:Anion Exchange Resins; D002852:Chromatography, Ion Exchange / Q000379:methods*; D003840:Deoxycholic Acid / Q000737:chemistry*; D018073:Haemophilus Vaccines / Q000737:chemistry*; D006851:Hydrochloric Acid / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D011135:Polysaccharides, Bacterial / Q000032:analysis*; D013745:Tetanus Toxoid / Q000737:chemistry*; D018074:Vaccines, Conjugate / Q000737:chemistry*
chemical_list: D000837:Anion Exchange Resins; D018073:Haemophilus Vaccines; C055753:Haemophilus influenza type b polysaccharide vaccine-tetanus toxin conjugate; D011135:Polysaccharides, Bacterial; D013745:Tetanus Toxoid; D018074:Vaccines, Conjugate; D003840:Deoxycholic Acid; D006851:Hydrochloric Acid