Monitoring bromide loss in bromoacetyl-derivatized polyribosylribitol polysaccharide in Haemophilus influenzae type b for PedvaxHIB® by capillary electrophoresis and NMR spectroscopy

Vaccine. 2022 Oct 6;40(42):6012-6016. doi: 10.1016/j.vaccine.2022.09.026. Epub 2022 Sep 17.

Abstract

PedvaxHIB® is an effective pediatric vaccine for protecting infants from invasive gram-negative bacterium Haemophilus influenzae type b. It is a highly purified capsular polysaccharide, polyribosylribitol phosphate that is covalently linked to an outer membrane protein complex of Neisseria meningitidis. PRP is first derivatized with an organic linker, followed by the coupling of a butadiamine group, and then at the end terminal, a bromoacetyl group is attached for conjugation with thiolated OMPC. The stability of the bromide group in derivatized PRP is monitored by two different methods, capillary electrophoresis and NMR spectroscopy. The loss of the bromide group is detected by measuring the amount of free bromide ion liberated using capillary electrophoresis and by observing a change in amide proton peaks near the bromide group using NMR. The two methods give similar rate hydrolysis results, therefore both can be employed as quick stability tools for bromoacetylation PRP content during manufacturing.

Keywords: Bromide loss kinetic; Capillary Electrophoresis; Haemophilus influenzae type b; NMR Spectroscopy; Polyribosylribitol phosphate polysaccharide; bromoacetylated-derivatized PRP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amides
  • Antibodies, Bacterial
  • Bacterial Outer Membrane Proteins
  • Bacterial Vaccines
  • Bromides
  • Child
  • Electrophoresis, Capillary
  • Haemophilus Infections* / prevention & control
  • Haemophilus Vaccines*
  • Haemophilus influenzae
  • Haemophilus influenzae type b*
  • Humans
  • Infant
  • Magnetic Resonance Spectroscopy
  • Phosphates
  • Polysaccharides, Bacterial
  • Protons

Substances

  • Amides
  • Antibodies, Bacterial
  • Bacterial Outer Membrane Proteins
  • Bacterial Vaccines
  • Bromides
  • Haemophilus Vaccines
  • Haemophilus influenzae-type b polysaccharide-Neisseria meningitidis outer membrane protein conjugate vaccine
  • Phosphates
  • Polysaccharides, Bacterial
  • Protons