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A scalable method for O-antigen purification applied to different Salmonella serovars

Micoli, Francesca, Rondini, Simona, Gavini, Massimiliano, Lanzilao, Luisa, Pisoni, Ivan, Colucci, Anna Maria, Sollai, Luigi, Giannelli, Carlo, Pippi, Federico, Maclennan, Calman Alexander, Pinto, Vittoria, Berti, Francesco, Saul, Allan James and Laura, Martin (2012) A scalable method for O-antigen purification applied to different Salmonella serovars. Analytical Biochemistry.


The surface lipopolysaccharide of Gram-negative bacteria is both a virulence factor and a B-cell antigen. Antibodies against O-antigen of lipopolysaccharide may confer protection against infection, and O-antigen conjugates have been designed against multiple pathogens. Here, we describe simplified methodology for extraction and purification of O-antigen-core portion of Salmonella lipopolysaccharide, suitable for large scale production. Lipopolysaccharide extraction and delipidation is performed by acetic acid hydrolysis of whole bacterial culture, and can take place directly in a bioreactor, without previous isolation and inactivation of bacteria. Further O-antigen-core purification consists of rapid filtration and precipitation steps, without using enzymes or hazardous chemicals. The process was successfully applied to different Salmonella enterica serovars (Paratyphi A, Typhimurium and Enteritidis), obtaining good yields of high quality material, suitable for conjugate vaccines preparations.

Item Type: Article
Date Deposited: 13 Oct 2015 13:14
Last Modified: 13 Oct 2015 13:14


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