VII Simposio Internacional de Química 2019 "SIQ 2019" -VII Conferencia "Ciencias Químicas"

VII Simposio Internacional de Química 2019

SIQ 2019

NMR FOR ASSESSING ASSOCIATED BIOMOLECULES FOCUSES ON ANTIBACTERIAL VACCINES

Resumen [ES]

Several Nuclear Magnetic Resonance (NMR) techniques has been created, adjusted or improved for analyzing aggregates and conjugates. These allow the access to different crucial parameters for assessing associated biomolecules. However, only few studies focused on OMV has been drived by NMR. This study is focused on the characterization of Neisseria meningititis (Nm) serogroup C capsular polysaccharide combined with OMV derived from Nm serogroup B and Salmonella enterica Typhi Vi capsular polysaccharide conjugated to Difteric toxoid (Se-Vi) by several NMR experiments. It comprises alternatives for assessing identity, size and their content. Sample comprises OMV prepared with outer membrane protein of serogroup B and capsular polysaccharide of serogroup C both from Nm. First, a 600 μL of OMV was mixing with 150 μL of a TSP-d4 / D2O solution (0.5 mg/mL). It yielded a solvent mixing of H2O/D2O : 80/20. Second, a aliquote of 600 μL was taken and added into a 5 mm NMR tube. Third, the quantitative proton NMR and Diffusion Order Spectroscopy (DOSY) spectra was acquired for the sample on two Bruker/Avance IV spectrometers one at 600 MHz and the other at 300 MHz. The identity of Nm and Se-Vi were achieved by checking their main fingerprints in the proton NMR spectrum. The content of these polysaccharides was also assessed by quantitative proton NMR spectrum. In addition, an alternative for studying self-diffusions rates in relation with polysaccharide average size was also illustrated. This work shows the possibility for studying capsular polysaccharides combined with OMV or conjugated by NMR.

Resumen [EN]

Several Nuclear Magnetic Resonance (NMR) techniques has been created, adjusted or improved for analyzing aggregates and conjugates. These allow the access to different crucial parameters for assessing associated biomolecules. However, only few studies focused on OMV has been drived by NMR. This study is focused on the characterization of Neisseria meningititis (Nm) serogroup C capsular polysaccharide combined with OMV derived from Nm serogroup B and Salmonella enterica Typhi Vi capsular polysaccharide conjugated to Difteric toxoid (Se-Vi) by several NMR experiments. It comprises alternatives for assessing identity, size and their content. Sample comprises OMV prepared with outer membrane protein of serogroup B and capsular polysaccharide of serogroup C both from Nm. First, a 600 μL of OMV was mixing with 150 μL of a TSP-d4 / D2O solution (0.5 mg/mL). It yielded a solvent mixing of H2O/D2O : 80/20. Second, a aliquote of 600 μL was taken and added into a 5 mm NMR tube. Third, the quantitative proton NMR and Diffusion Order Spectroscopy (DOSY) spectra was acquired for the sample on two Bruker/Avance IV spectrometers one at 600 MHz and the other at 300 MHz. The identity of Nm and Se-Vi were achieved by checking their main fingerprints in the proton NMR spectrum. The content of these polysaccharides was also assessed by quantitative proton NMR spectrum. In addition, an alternative for studying self-diffusions rates in relation with polysaccharide average size was also illustrated. This work shows the possibility for studying capsular polysaccharides combined with OMV or conjugated by NMR.

Sobre el ponente

Raine Garrido Arteaga

Dr. Raine Garrido Arteaga

IFV Flag of Cuba
Información Práctica
English (US)
No definido
30 minutos
No definido
Autores
Dr. Raine Garrido Arteaga
Eddy Petit
José Luis Pérez
Bárbara Baró
Françoise Keita Paquet
Palabras clave
analysis
interactions
nmr
vesicles