Abstract
To facilitate the routine identification of salmonellae and detailed studies of their lipopolysaccharides, we raised murine monoclonal antibodies against these organisms. We raised an immunoglobulin G1 antibody, MO2, which is specific for factor O2. By immunoblotting following sodium dodecyl sulfate-polyacrylamide gel electrophoresis, MO2 was shown to bind only the lipopolysaccharide of Salmonella paratyphi A, giving a ladderlike reaction pattern with regularly spaced reactive bands. MO2 did not react against lipopolysaccharides of other Salmonella serogroups, including those of serogroups B, C, D, E, and L. Since the lipopolysaccharides of Salmonella serogroups A, B, D, and E are similar except for the presence of paratose in serogroup A organisms, this dideoxyhexose is therefore believed to be the immunodominant epitope for MO2. Consistent with the latter contention was the finding that periodate oxidation of the S. paratyphi A lipopolysaccharide did not destroy its antigenicity for MO2. In a slide agglutination test, MO2 was found to react specifically against all 12 clinical isolates of S. paratyphi A but not against 98 isolates of other salmonellae or 74 isolates of other bacteria and Candida albicans.
MeSH Terms
Animals
Antibodies, Monoclonal/immunology
Antigens, Bacterial/immunology
Electrophoresis, Polyacrylamide Gel
Enzyme-Linked Immunosorbent Assay
Epitopes/analysis,immunology
Hybridomas
Immunoassay
Lipopolysaccharides/immunology
Mice
O Antigens
Salmonella paratyphi A/immunology
Chemicals
Antibodies, Monoclonal
Antigens, Bacterial
Epitopes
Lipopolysaccharides
O Antigens
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Luk M C
Department of Microbiology, University of Hong Kong.
Tsang R S
Ng M H
References (11)
11 references, click to expand
-
Enzyme-linked immunosorbent assay, Elisa. 3. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes.
J Immunol. 1972 Jul;109(1):129-35
PMID: 4113792
-
Continuous cultures of fused cells secreting antibody of predefined specificity.
Nature. 1975 Aug 7;256(5517):495-7
PMID: 1172191
-
Heterogeneity of lipopolysaccharides. Analysis of polysaccharide chain lengths by sodium dodecylsulfate-polyacrylamide gel electrophoresis.
Eur J Biochem. 1975 Dec 1;60(1):239-46
PMID: 1107034
-
Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4
PMID: 388439
-
Heterogeneity of antigenic-side-chain length in lipopolysaccharide from Escherichia coli 0111 and Salmonella typhimurium LT2.
Eur J Biochem. 1980;107(1):145-53
PMID: 6156828
-
Protein measurement with the Folin phenol reagent.
J Biol Chem. 1951 Nov;193(1):265-75
PMID: 14907713
-
Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels.
J Bacteriol. 1983 Apr;154(1):269-77
PMID: 6187729
-
Detection of monoclonal antibodies specific for carbohydrate epitopes using periodate oxidation.
J Immunol Methods. 1985 Apr 8;78(1):143-53
PMID: 2580026
-
Rapid serological analysis of bacterial lipopolysaccharides by electrotransfer to nitrocellulose.
J Immunol Methods. 1985 Dec 27;85(2):371-82
PMID: 3908569
-
A murine monoclonal antibody specific for the outer core oligosaccharide of Salmonella lipopolysaccharide.
Infect Immun. 1987 Jan;55(1):211-6
PMID: 2432013
-
A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels.
Anal Biochem. 1982 Jan 1;119(1):115-9
PMID: 6176137