Column chromatography and cell culture assay of Pseudomonas aeruginosa toxin Z preparations.
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.
D000818:Animals; D000907:Antibodies, Bacterial; D001681:Biological Assay; D002460:Cell Line; D002850:Chromatography, Gel; D006801:Humans; D007118:Immunoassay; D009500:Neutralization Tests; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D011817:Rabbits; D014118:Toxins, Biological / Q000096:biosynthesis / Q000302:isolation & purification* / Q000494:pharmacology
D000907:Antibodies, Bacterial; D014118:Toxins, Biological