Size, charge and structural heterogeneity of Brucella abortus lipopolysaccharides demonstrated by two‐dimensional gel electrophoresis

Phenol extracted, alkali‐treated lipopolysaccharide (aLPS) from vaccine strain (S19) Brucella abortus was demonstrated by two‐dimensional gel electrophoresis to consist of at least ten silver staining, polydisperse analogues having different pIs. When tested on nitrocellulose immunoblots, all ten were antigenically reactive with bovine anti‐B. abortus polyclonal sera, but only six reacted with anti‐B. abortus O‐antigen murine monoclonal antibody. Analogues focusing at different pIs were concluded to arise from differences in either core or O‐antigen side chain structure or because of covalently bound protein. While not qualitatively different, aLPS from pathogenic B. abortus strain 2308 had lesser amounts of analogues 1, 2, 5, 6, and 8 than did aLPS from strain 19 (vaccine). The 2‐D gel electrophoresis method was demonstrated to be of value in the analysis of aLPS from B. abortus and may be useful in the study of lipopolysaccharides from other sources.

[1]  M. Perry,et al.  Antigenic S-type lipopolysaccharide of Brucella abortus 1119-3 , 1984, Infection and immunity.

[2]  A. M. Wu,et al.  Immunochemical studies on the binding properties of Brucella abortus lipopolysaccharides to bovine precipitating antibodies. , 1984, Molecular immunology.

[3]  T. Marshall,et al.  Artifacts associated with 2-mercaptoethanol upon high resolution two-dimensional electrophoresis. , 1984, Analytical biochemistry.

[4]  B. Ganem,et al.  Evidence for covalent bonding of native hapten–protein complexes to smooth lipopolysaccharide of Brucella abortus , 1984 .

[5]  J. Katzmann,et al.  Development of monoclonal antibodies to proteins separated by two-dimensional gel electrophoresis. , 1983, Journal of immunological methods.

[6]  D. Ochs,et al.  Protein contaminants of sodium dodecyl sulfate-polyacrylamide gels. , 1983, Analytical biochemistry.

[7]  I. Poxton,et al.  Analysis of lipopolysaccharides of Bacteroides fragilis by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and electroblot transfer , 1983 .

[8]  H. D. Hochstein,et al.  Physical and biological properties of U.S. standard endotoxin EC after exposure to ionizing radiation , 1983, Infection and immunity.

[9]  K. Nielsen,et al.  Interaction of specifically purified isotypes of bovine antibody to Brucella abortus in the haemolysis in gel test and enzyme-linked immunosorbent assay. , 1983, Research in veterinary science.

[10]  K. Nielsen,et al.  Derivation of monoclonal antibodies against Brucella abortus antigens. , 1983, Veterinary immunology and immunopathology.

[11]  P. Hitchcock Aberrant migration of lipopolysaccharide in sodium dodecyl sulfate/polyacrylamide gel electrophoresis. , 1983, European journal of biochemistry.

[12]  K. Nielsen,et al.  Affinity purification of bovine antibodies to Brucella abortus Lipopolysaccharide , 1983, Journal of clinical microbiology.

[13]  B. Sowa,et al.  Physiology of F-Pilin Synthesis and Utilization , 1983, Journal of bacteriology.

[14]  B. L. Deyoe,et al.  Antibodies to Brucella abortus in sera from Strain 19 vaccinated and non-vaccinated cows as determined by enzyme linked immunosorbent assay and conventional serologic methods. , 1982, Veterinary immunology and immunopathology.

[15]  C. Boulard,et al.  Specific antisera produced by direct immunization with slices of polyacrylamide gel containing small amounts of protein. , 1982, Journal of immunological methods.

[16]  R. Munford,et al.  Size heterogeneity of Salmonella typhimurium lipopolysaccharides in outer membranes and culture supernatant membrane fragments , 1980, Journal of bacteriology.

[17]  L. Leive,et al.  Heterogeneity of antigenic-side-chain length in lipopolysaccharide from Escherichia coli 0111 and Salmonella typhimurium LT2. , 1980, European journal of biochemistry.

[18]  P. Mäkelä,et al.  Lipopolysaccharide heterogeneity in Salmonella typhimurium analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. , 1980, European journal of biochemistry.

[19]  L. Anderson,et al.  Analytical techniques for cell fractions. XXVIII. Dissection of complex antigenic mixtures using monoclonal antibodies and two-dimensional gel electrophoresis. , 1980, Analytical biochemistry.

[20]  H. Towbin,et al.  Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[21]  G. Schurig,et al.  Purification and characterization of smooth and rough lipopolysaccharides from Brucella abortus , 1979, Journal of bacteriology.

[22]  R. E. Hurlbert,et al.  Biological and physicochemical properties of the lipopolysaccharide of Chromatium vinosum , 1977, Infection and immunity.

[23]  R. Russell,et al.  SDS-polyacrylamide gel electrophoresis of lipopolysaccharides. , 1975, Canadian journal of microbiology.

[24]  K. Jann,et al.  Heterogeneity of lipopolysaccharides. Analysis of polysaccharide chain lengths by sodium dodecylsulfate-polyacrylamide gel electrophoresis. , 1975, European journal of biochemistry.

[25]  P. O’Farrell High resolution two-dimensional electrophoresis of proteins. , 1975, The Journal of biological chemistry.

[26]  A. Olins,et al.  Physicochemical studies on a lipopolysaccharide from the cell wall of Azotobacter vinelandii. , 1967, The Journal of biological chemistry.

[27]  P. Baker,et al.  Hypoferremia in Mice and Its Application to the Bioassay of Endotoxin , 1965, Journal of bacteriology.

[28]  L. Hood,et al.  Isolation of microgram quantities of proteins from polyacrylamide gels for amino acid sequence analysis. , 1983, Methods in enzymology.

[29]  C. Frasch,et al.  A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. , 1982, Analytical biochemistry.