使用识别分枝菌酸的多克隆抗体对形成浮渣细菌进行免疫磁分离。

Immunomagnetic separation of scum-forming bacteria using polyclonal antibody that recognizes mycolic acids.

作者信息

Morisada S, Miyata N, Iwahori K

机构信息

Graduate School of Nutritional and Environmental Sciences, University of Shizuoka, 52-1 Yada, 422-8526, Shizuoka, Japan.

出版信息

J Microbiol Methods. 2002 Oct;51(2):141-8. doi: 10.1016/s0167-7012(02)00046-5.

DOI:10.1016/s0167-7012(02)00046-5

PMID:12133606

Abstract

Mycolic acid-containing bacteria (mycolata) are thought to be involved in scum formation in aeration basins of activated sludge plants due to their ability to produce biosurfactants and their cell surface hydrophobicity. To isolate these bacteria, immunomagnetic separation (IMS) using an anti-mycolic acid polyclonal antibody was investigated. IMS that targeted Gordonia amarae SC1 exhibited a 100% recovery at 5x10(3) CFU ml(-1). At cell concentration of 7.8x10(6) CFU ml(-1), the recovery was lowered, but 80% of cells were still captured. Effect of bead concentrations on the recovery of SC1 at 10(6) CFU ml(-1) was examined. The results showed that addition of more than 6-7x10(6) beads for 1x10(6) CFU reached a maximum recovery (83%). Furthermore, the IMS procedure optimized with SC1 cells was tested with another mycolata. The results suggested that variation of the recovery for each mycolata is dependent on the specificity of the polyclonal antibody and that mycolata which are recognized by the antibody can be recovered by this procedure.

摘要

含分枝菌酸的细菌（分枝杆菌纲）被认为由于其产生生物表面活性剂的能力及其细胞表面疏水性，而与活性污泥厂曝气池中浮渣的形成有关。为了分离这些细菌，研究了使用抗分枝菌酸多克隆抗体的免疫磁珠分离法（IMS）。针对戈氏菌SC1的免疫磁珠分离法在5×10³CFU/ml时回收率为100%。在细胞浓度为7.8×10⁶CFU/ml时，回收率降低，但仍有80%的细胞被捕获。研究了磁珠浓度对10⁶CFU/ml的SC1回收率的影响。结果表明，对于1×10⁶CFU，加入超过6 - 7×10⁶个磁珠时回收率达到最大值（83%）。此外，用另一种分枝杆菌纲细菌测试了用SC1细胞优化的免疫磁珠分离法程序。结果表明，每种分枝杆菌纲细菌回收率的差异取决于多克隆抗体的特异性，并且被该抗体识别的分枝杆菌纲细菌可以通过此程序回收。