Column chromatography and cell culture assay of Pseudomonas aeruginosa toxin Z preparations
- PMID: 809009
- PMCID: PMC187170
- DOI: 10.1128/am.30.2.293-297.1975
Column chromatography and cell culture assay of Pseudomonas aeruginosa toxin Z preparations
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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