Mice immunization with gel electrophoresis-micropurified bacterial lipopolysaccharides

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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 μm 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.