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Salmonella Typhimurium

(later renamed to: Salmonella enterica ssp. enterica sv. Typhimurium)
(NCBI TaxID 90371, species name lookup)

Taxonomic group: bacteria / Proteobacteria (Phylum: Proteobacteria)
Host organism: Homo sapiens
Associated disease: bacteremia [ICD11: MA15.0

]; infection due to Salmonella enterica [ICD11: XN5VC

]

NCBI PubMed ID: 23142459
Publication DOI: 10.1016/j.jim.2012.10.015
Journal NLM ID: 1305440
Correspondence: C.A. MacLennan <calman.maclennan

novartis.com>
Institutions: Novartis Vaccines Institute for Global Health, Via Fiorentina 1, 53100 Siena, Italy, Medical Research Council Centre for Immune Regulation and Clinical Immunology Service, Institute of Biomedical Research, School of Immunity and Infection, College of Medicine and Dental Sciences, University of Birmingham, Birmingham, UK

Nontyphoidal Salmonellae (NTS) are a common cause of bacteraemia in children and HIV-infected adults in Sub-Saharan Africa. We have previously shown that antibodies play a key role in both bactericidal and cellular mechanisms of immunity to NTS, but found that high concentrations of antibody to Salmonella Typhimurium O antigen (OAg) in the serum of some HIV-infected African adults is associated with impaired killing of NTS. To further investigate the function of antibodies to the OAg NTS, we developed a method to purify these antibodies from human serum by affinity chromatography. Purified Salmonella Typhimurium OAg was activated with adipic acid dihydrazide (ADH) via two different chemistries before linking to N-hydroxysuccinamide-Sepharose resin: one ADH molecule was introduced per OAg chain on its terminal 3-deoxy-D-manno-octulosonic acid sugar (OAg-ADH), or multiple ADH molecules were attached along the OAg chain after oxidation with sodium periodate (OAgoxADH). Both resulting columns worked well when tested with commercial polyclonal anti-O:4,5 antibodies from rabbit serum. Over 90% of the applied antibodies bound to the resin and 89% of the linked antibodies were then eluted as detected by ELISA. OAg-ADH was preferred as the method for OAg derivatisation as it does not modify the saccharide chain and can be applied to OAg from different bacteria. Both columns were able to bind OAg-specific antibodies in human serum, but antibody recovery was initially low. Different elution buffers were tested and different amounts of OAg-ADH were linked to the resin to improve the yield. Optimal recovery (51%) was obtained by loading 1mg of activated OAg per ml of resin and eluting with 0.1M glycine, 0.1M NaCl pH2.4. The column matrix could be regenerated following elution with no detectible loss in performance for over ten uses. This method offers the potential to purify antibodies to Salmonella OAg from polyclonal serum following vaccination or natural exposure to Salmonella and so investigate the functionality and diversity of the antibody response to OAg.

O-antigen, antibody, Salmonella, serum, purification, affinity

Structure type: oligomer
Location inside paper: p.201, fig.1A
Compound class: core oligosaccharide
Contained glycoepitopes: IEDB_120354,IEDB_123890,IEDB_130650,IEDB_130693,IEDB_136906,IEDB_137340,IEDB_137472,IEDB_140529,IEDB_141794,IEDB_141807,IEDB_142488,IEDB_144998,IEDB_146664,IEDB_151528,IEDB_151531,IEDB_167069,IEDB_190606,IEDB_983931,SB_192,SB_7

Methods: 1H NMR, ELISA, serological methods, HPLC
Biological activity: the purification of antibodies specific to S. Typhimurium OAg from human serum by affinity chromatography.
Comments, role: the OAg chain repeating unit (ID 29296) is attached the core region

Related record ID(s): 29296
NCBI Taxonomy refs (TaxIDs): 90371
Show glycosyltransferases

There are 2 chemically distinct structures. Please, select:

[1*][3*](OC1O[C@H](CO[C@H]2O[C@H](CO)[C@H](O)[C@H](O)[C@H]2O)[C@@H](O)[C@H](O[C@H]2O[C@H](CO)[C@H](O)[C@H](O)[C@H]2O[C@H]2O[C@H](CO)[C@@H](O[2*])[C@H](O)[C@H]2O[C@H]2O[C@H](CO)[C@@H](O)[C@H](O)[C@H]2NC(C)=O)[C@H]1O)(OP(=O)(O)O)[3*](O[C@@H]1[C@H](O)CC(O)(C(=O)O)O[C@@H]1[C@H](O)CO)OP(=O)(O)OP(=O)(O)OCCN

SVG MW SMILES 3D mol Ionize

R1 = ??Hepp
R2 = O-antigen (ID 29296)
R3 = a?Hepp