铜绿假单胞菌毒素Z制剂的柱色谱分析及细胞培养测定

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

作者信息

Ludovici P P, Roinestad F A, Wong F S

出版信息

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

DOI:10.1128/am.30.2.293-297.1975

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC187170/

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.

摘要

铜绿假单胞菌在细胞培养中产生的有毒物质被浓缩并部分纯化。这种有毒物质被命名为毒素Z，是菌株PA Z或PA 103在含有10%血清的伊格尔基础培养基的HEp-2单层培养物生长过程中产生的。通过Lyphogel或超滤浓缩四倍的毒素Z用于在兔体内产生抗血清，并且还通过柱色谱进行分级分离。在Sephadex G - 200柱上实现了毒素Z的20倍纯化。通过用特异性抗血清中和来确认有毒柱级分含有毒素Z。在浓缩、纯化和中和过程中，毒素仅通过其在细胞培养中产生的细胞病变效应进行测定。

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