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Column Chromatography and Cell Culture Assay of Pseudomonas aeruginosa Toxin Z Preparations
Authors:Peter P. Ludovici   Frank A. Roinestad     Francis S. Wong
Abstract:Toxic material produced by Pseudomonas aeruginosa in cell culture was concentrated and partially purified. This toxic material, designated toxin Z, was produced during the growth of strain PA Z or PA 103 in HEp-2 monolayer cultures using Eagle minimal essential medium with 10% serum. Toxin Z, concentrated fourfold by Lyphogel or ultrafiltration, was used to produce antiserum in rabbits and also was fractionated by column chromatography, Twentyfold purification of toxin Z was obtained on a Sephadex G-200 column. Toxic column fractions were confirmed to have toxin Z by neutralization with specific antiserum. During concentration, purification, and neutralization procedures, the toxin was assayed exclusively by the cytopathic effect it produced in cell culture.
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