Mice immunization with gel electrophoresis-micropurified bacterial lipopolysaccharides.
Some evidence on the possible use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to elicit antibodies against smooth- or rough-type bacterial lipopolysaccharides (LPS) is shown. Gel-separated LPS were negatively stained with zinc-imidazole to precisely localize the bands of interest under fully reversible conditions. Then the bands of interest were excised and the resulting gel slices washed in a solution of a zinc-complexing agent (e.g., 100 mM EDTA), after which they were extruded through a metal sieve of 32 microm average size contained in a 1 mL syringe, to generate homogeneous gel microparticles. The LPS-containing gel slurries were used directly to immunize female BALB/c mice. Using this procedure, positive mouse polyclonal antibody responses against gel-purified smooth- or rough-LPS forms from Escherichia coli K-235 or Bordetella pertussis were elicited, as tested by a dot-immunoblotting assay. Our results may encourage the use of SDS-PAGE-micropurified LPS to develop optimized immunization procedures for the generation of specific antibodies against LPS bands of defined sizes, and therefore they constitute an intermediate step toward that aim.
Pupo
E
E
Center for Genetic Engineering and Biotechnology, Havana, Cuba.
Aguila
A
A
Santana
H
H
Núñez
J F
JF
Castellanos-Serra
L
L
Hardy
E
E
D000818:Animals; D001886:Bordetella pertussis / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004926:Escherichia coli / Q000276:immunology; D005260:Female; D008070:Lipopolysaccharides / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D014611:Vaccination
D008070:Lipopolysaccharides