McElhiney J, Lawton L A, Porter A J
Department of Molecular and Cell Biology, Institute of Medical Sciences, University of Aberdeen, Foresterhill, UK.
FEMS Microbiol Lett. 2000 Dec 1;193(1):83-8. doi: 10.1111/j.1574-6968.2000.tb09406.x.
Single-chain antibody fragments against the cyanobacterial hepatotoxin microcystin-LR were isolated from a naive human phage display library and expressed in Escherichia coli. In competition enzyme-linked immunosorbent assay (ELISA), the most sensitive antibody clone selected from the library detected free microcystin-LR with an IC(50) value of 4 microM. It was found to cross react with three other microcystin variants - microcystin-RR, microcystin-LW and microcystin-LF - and detected microcystins in extracts of the cyanobacterium Microcystis aeruginosa, found to contain the toxins by high-performance liquid chromatography (HPLC). The quantification of microcystins in these extracts by ELISA and HPLC showed good correlation. Although the antibody isolated in this study was considerably less sensitive than the polyclonal and monoclonal antibodies already available for microcystin detection, phage display technology represents a cheaper, more rapid alternative for the production of anti-microcystin antibodies than the methods currently in use.
从一个未经免疫的人噬菌体展示文库中分离出了针对蓝藻肝毒素微囊藻毒素-LR的单链抗体片段，并在大肠杆菌中进行了表达。在竞争酶联免疫吸附测定（ELISA）中，从文库中选出的最敏感的抗体克隆检测游离微囊藻毒素-LR的IC(50)值为4微摩尔。发现它与其他三种微囊藻毒素变体——微囊藻毒素-RR、微囊藻毒素-LW和微囊藻毒素-LF——发生交叉反应，并在铜绿微囊藻提取物中检测到微囊藻毒素，通过高效液相色谱法（HPLC）发现该提取物含有这些毒素。通过ELISA和HPLC对这些提取物中的微囊藻毒素进行定量分析显示出良好的相关性。尽管本研究中分离出的抗体比目前可用于微囊藻毒素检测的多克隆和单克隆抗体的敏感性要低得多，但噬菌体展示技术是一种比目前使用的方法更便宜、更快速的生产抗微囊藻毒素抗体的替代方法。
