Method for producing a vaccine against bacterial infections caus

Drug – bio-affecting and body treating compositions – Conjugate or complex of monoclonal or polyclonal antibody,... – Conjugated via claimed linking group – bond – chelating agent,...

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424 88, 424 92, 536 1, A61K 3170, C08B 3700, A61K 39104

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active

042859360

ABSTRACT:
A method for isolating a non toxic, high molecular weight polysaccharide igen from the crude slime of a Pseudomonas aeruginosa culture, and a method for inducing immunity in a host to said live organisms is described. The slime is initially prepared for the isolation procedure by separating the bacterial cells from the slime and dissolving the slime in a phosphate buffer solution. Dissolved contaminating nucleic acids are then precipitated and separated from solution. A lipid A portion of the contaminating lipopolysaccharide constituent is then removed and precipitated by acetic acid hydrolysis, and the remaining lipids present are extracted with chloroform. Nearly all of the residual nucleic acids are then removed by digestion with nucleases, and the remaining protein extracted with phenol. The aqueous and phenol layers are then separated, and the aqueous layer applied to a gel filter to isolate the antigen by column chromatography. The antigen appears in the void volume. A vaccine containing the polysaccharide antigen of this invention has been successfully tested and found to be non toxic and highly effective in inducing an immune response to the organism in a host.

REFERENCES:
Pier, G., et al., Injection and Immunity, vol. 22, pp. 908-925, 1978.

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