"Isolation of O-polysaccharide from the cell wall LPS does not represent significant technical difficulties. Thus, the method of isolation, first proposed by Freeman, includes the following main stages--obtaining culture of bacteria S. sonnei, phase I in liquid medium; separation of culture fluid from bacterial cells, extracting LPS from bacterial cell with aqueous phenol (Westphal O.,
"Another method of obtaining highly purified O-specific antigen of Shigella sp is also known and includes the following stages: obtaining bacterial cultures in liquid medium; treatment of bacterial cultures with hexadecyltrimethylammonium bromide and subsequent extraction of LPS from bacterial cells; separation of LPS extract from bacterial cells; degradation of LPS with subsequent separation of O-polysaccharide (KR 20010054032 A). Thereby, all known methods of isolating O-specific antigens from Shigella sp. LPS are based on the stage of extraction, i.e. LPS extraction from bacterial cells, which causes the unavoidable loss of bacterial cell nativity.
"It should be additionally marked, that the structure of O-specific antigens obtained by known methods from LPS's is determined by genomes of Shigella sp bacteria.
"Practically all O-antigens obtained from Shigella sp. LPS's contain elements of core domain structures. Mild hydrolysis using 1% acetic acid is used for removal of lipid A from the LPS molecule, leads to obtaining a polysaccharide derivative, which is represented as a O-specific polysaccharide, connected to the 'core' oligosaccharide (Fensom A. H.,
"It was proposed to use the O-polysaccharide from the LPS of the bacterial cell wall of S. sonnei, phase I, as a component of only conjugated vaccines against S.sonnei shigellosis, under it's covalent bonding with protein carriers--protein D Haemophilis influenzae, recombinant exoprotein A Pseudomonas aeruginosa (rEPA), recombinant diphtheria toxin (rDT), recombinant toxin B Clostriduum. difficile (rBRU) (US Pat. Appl. 2005/0031646; WO/2010/019890).
"Investigations were conducted of the immunogenic and protective properties of conjugates containing O-polysaccharide from the LPS of the bacterial cells of Plesiomonas shigelloides O7, whose structure is identical to O-polysaccharide from LPS of bacteria S. sonnei, phase I, and proteins--exoprotein A P. aeruginosa (rEPA) or diphtheria toxoid CRM9 from mutant strain Corynobacterium diphtheriae (Cohen D., Ashkenazi S., Green M. S., Gdalevich M., Robin G., Slepon R., Yavzori M., Orr N., Block C., Ashkenazi I., Shemer J.,
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