The polysaccharides of interest have been analyzed by one or more of a variety of NMR methods, including 1D 1H NMR, 2D correlation spectroscopy (COSY), and 2D total correlation spectroscopy (TOCSY) for assignment of 1H spectra, and 1D 13C NMR, 1H coupled and decoupled, 2D heteronuclear single quantum correlation (HSQC), and 2D heteronuclear multiple bond correlation (HMBC) for the elucidation of 13C NMR spectra.? ? NEISSERIA MENINGITIDIS W135: Preparations of native and de-O-acetylated capsular polysaccharide from W135 CPS were analyzed for N-acetyl and O-acetyl content by 1H NMR spectroscopy. Quantitative integration of the N-acetyl and O-acetyl methyl signals of the 7-O-acetyl- and 9-O-acetyl-N-acetylneuraminic acid components indicated that the total O-acetyl content of the CPS was 1.1-1.2 mole%, i.e., close to the detection limit of the spectrometer.? ? HAEMOPHILUS INFLUENZAE A: Preparations of the native and purified CPS were analyzed by 1D 1H NMR, 1D 13C NMR, 2D COSY, 2D TOCSY, 1H coupled and decoupled 2D HSQC, and 2D HMBC. The strongest evidence for the polyphosphate structure of the CPS was apparent in the 13C NMR spectrum, which displayed five 13C-31P coupling constants in the appropriate 13C resonances. NMR also indicated that the purity of the preparations was excellent.
| Jin, Zhigang; Bohach, Gregory A; Shiloach, Joseph et al. (2005) Conjugates of group A and W135 capsular polysaccharides of neisseria meningitidis bound to recombinant Staphylococcus aureus enterotoxin C1: preparation, physicochemical characterization, and immunological properties in mice. Infect Immun 73:7887-93 |
