title: Mice immunization with gel electrophoresis-micropurified bacterial lipopolysaccharides.
abstract: 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.
authors.affiliation: Center for Genetic Engineering and Biotechnology, Havana, Cuba.
authors.forename: E
authors.initials: E
authors.lastname: Pupo
authors.forename: A
authors.initials: A
authors.lastname: Aguila
authors.forename: H
authors.initials: H
authors.lastname: Santana
authors.forename: J F
authors.initials: JF
authors.lastname: Núñez
authors.forename: L
authors.initials: L
authors.lastname: Castellanos-Serra
authors.forename: E
authors.initials: E
authors.lastname: Hardy
mesh_terms: 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
chemical_list: D008070:Lipopolysaccharides