Quantitative nuclear magnetic resonance analysis and characterization of the derivatized Haemophilus influenzae type b polysaccharide intermediate for PedvaxHIB.
PedvaxHIB is a pediatric vaccine that protects children from severe disease caused by the gram-negative bacterium Haemophilus influenzae type b (Hib). The vaccine is made by chemically conjugating Hib capsular polysaccharide to the outer membrane protein complex of Neisseria meningitidis. The protein-conjugated vaccine has proven to be extremely effective in preventing invasive Hib disease in infants and young children. This paper presents the nuclear magnetic resonance (NMR) methodology for the quantitative characterization of derivatized polysaccharide and its validation closely following ICH guidelines. The assay has been shown to be precise and accurate (relative standard deviation [RSD]<or=1%), specific (no observable matrix interference), rugged (RSD <3% for day-to-day and operator-to-operator variations), and robust (to changes in temperature and polysaccharide concentration). Therefore, the quantitative NMR assay can be used as a reliable product release or process monitoring method and can replace several labor-intensive chromatographic and colorimetric methods.
Xu
Qiuwei
Q
Merck Research Laboratory, Merck & Co., Inc., West Point, PA 19486, USA. qiuwei_xu@merck.com
Klees
Jennifer
J
Teyral
Jennifer
J
Capen
Robert
R
Huang
Michael
M
Sturgess
Annie W
AW
Hennessey
John P
JP
Washabaugh
Michael
M
Sitrin
Robert
R
Abeygunawardana
Chitrananda
C
D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry*; D018073:Haemophilus Vaccines / Q000737:chemistry*; D019741:Haemophilus influenzae type b / Q000737:chemistry* / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D011135:Polysaccharides, Bacterial / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013379:Substrate Specificity; D018074:Vaccines, Conjugate / Q000737:chemistry
D001425:Bacterial Outer Membrane Proteins; D018073:Haemophilus Vaccines; C061964:Haemophilus influenzae-type b polysaccharide-Neisseria meningitidis outer membrane protein conjugate vaccine; D011135:Polysaccharides, Bacterial; D018074:Vaccines, Conjugate