用于纯化溶细胞性细菌毒素的疏水作用色谱法。

Hydrophobic interaction chromatography for the purification of cytolytic bacterial toxins.

作者信息

Schoel B, Welzel M, Kaufmann S H

机构信息

Department of Immunology, University of Ulm, Germany.

出版信息

J Chromatogr A. 1994 Apr 29;667(1-2):131-9. doi: 10.1016/0021-9673(94)89061-7.

DOI:10.1016/0021-9673(94)89061-7

PMID:8025627

Abstract

The usefulness of hydrophobic interaction chromatography for the simple purification of cytolytic bacterial toxins was studied. Conditions are described for different hydrophobic interaction chromatographic media for purifying with high yields two different kinds of such haemolysins, the thiol-activated toxin listeriolysin O from Listeria monocytogenes and alpha-toxin from Staphylococcus aureus. For listeriolysin O, purification on butyl-Sepharose was followed by gel filtration chromatography. From butyl-Sepharose the recovery of 22%. Alpha-toxin was obtained by a single purification step from alkyl-Superose with 80% recovery and a specific activity of 29,000 U/mg. On sodium dodecyl sulphate polyacrylamide gel electrophoresis purified listeriolysin O and alpha-toxin showed a single band. Another thiol-activated toxin, streptolysin O from group A streptococci, showed a recovery of 38% from butyl-Sepharose. The results suggest the feasibility of using hydrophobic interaction chromatography, particularly with columns of weak hydrophobicity, for the purification of bacterial haemolysins in high yield.

摘要

研究了疏水作用色谱法用于简单纯化溶细胞性细菌毒素的实用性。描述了使用不同疏水作用色谱介质的条件，以便高产量地纯化两种不同的溶血素，即来自单核细胞增生李斯特菌的硫醇激活毒素李斯特菌溶血素O和来自金黄色葡萄球菌的α毒素。对于李斯特菌溶血素O，在丁基琼脂糖凝胶上纯化后再进行凝胶过滤色谱。从丁基琼脂糖凝胶上的回收率为22%。α毒素通过在烷基超级琼脂糖上进行单一纯化步骤获得，回收率为80%，比活性为29,000 U/mg。在十二烷基硫酸钠聚丙烯酰胺凝胶电泳上，纯化后的李斯特菌溶血素O和α毒素显示为单一条带。另一种硫醇激活毒素，来自A组链球菌的链球菌溶血素O，从丁基琼脂糖凝胶上的回收率为38%。结果表明，使用疏水作用色谱法，特别是使用弱疏水性柱，以高产量纯化细菌溶血素是可行的。