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. 1975 Aug;30(2):293-7.
doi: 10.1128/am.30.2.293-297.1975.

Column chromatography and cell culture assay of Pseudomonas aeruginosa toxin Z preparations

P P Ludovici, F A Roinestad, F S Wong

Column chromatography and cell culture assay of Pseudomonas aeruginosa toxin Z preparations

P P Ludovici et al. Appl Microbiol. 1975 Aug.

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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