Multiple Techniques for Size Determination of Generalized Modules for Membrane Antigens from Salmonella typhimurium and Salmonella enteritidis

ACS Omega
Gianluigi De BenedettoF Micoli

Abstract

In the last years, outer membrane vesicles have attracted a lot of attention for the development of vaccines against bacterial pathogens. Extracellular vesicles can be obtained in high yields by genetic mutations, resulting in generalized modules for membrane antigens (GMMA). Methods to check the quality, consistency of production, and stability of GMMA vaccines are of fundamental importance. In this context, analytical methods for size distribution determination and verifying the integrity and possible aggregation of GMMA particles are strongly needed. Herein, GMMA particle size distribution has been evaluated by means of three different techniques. Dynamic light scattering (DLS), multiangle light scattering (MALS) coupled with high-performance liquid chromatography-size exclusion chromatography (SEC), and nanoparticle tracking analysis (NTA) have been compared to characterize GMMA from different mutants of Salmonella typhimurium and Salmonella enteritidis strains. We found that the presence of O-antigen chains on GMMA determined higher Z-average diameters by DLS compared to size estimation by MALS and that the hydrodynamic diameter increased with the number of O-antigen chains per GMMA particle. In the case of SEC-MALS, the s...Continue Reading

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Citations

Sep 29, 2018·Proceedings of the National Academy of Sciences of the United States of America·Francesca MicoliCalman A MacLennan
Aug 4, 2021·Physical Chemistry Chemical Physics : PCCP·Fabian SoltermannPhilipp Kukura
Apr 27, 2018·Bioconjugate Chemistry·Gianluigi De BenedettoFrancesca Micoli

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Methods Mentioned

BETA
acylation
dynamic light scattering
light scattering
static light scattering
light-scattering
glucosylation
Protein Assay

Software Mentioned

Zetasizer
ASTRA
GMMA
MALS
SEC
NTA

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