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针对鸟分枝杆菌特定血清型以及鸟分枝杆菌-胞内分枝杆菌-瘰疬分枝杆菌复合群的单克隆抗体的制备与特性分析

Production and characterization of monoclonal antibodies against specific serotypes of Mycobacterium avium and the Mycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceum complex.

作者信息

Kolk A H, Evers R, Groothuis D G, Gilis H, Kuijper S

机构信息

N. H. Swellengrebel Laboratory of Tropical Hygiene, Royal Tropical Institute, Amsterdam, The Netherlands.

出版信息

Infect Immun. 1989 Aug;57(8):2514-21. doi: 10.1128/iai.57.8.2514-2521.1989.

Abstract

Serotype-specific and Mycobacterium avium-Mycobacterium intracellulare-Mycobacterium scrofulaceum complex (MAIS complex)-specific monoclonal antibodies (MAbs) were prepared. A series of MAbs were obtained, five specific for serotype 2, three specific for serotype 4, eight against a strain of serotype 19, two specific for the MAIS complex, and two against a common glycolipid shared by all the mycobacteria tested so far. The serotype-specific and the MAIS complex-specific MAbs reacted in immunofluorescence with intact mycobacteria and in enzyme-linked immunosorbent assay and immuno-thin-layer chromatography with lipid extracts of mycobacteria. The two MAbs against a common mycobacterial glycolipid reacted only in lipid enzyme-linked immunosorbent assay and immuno-thin-layer chromatography. All MAbs were directed against glycopeptidolipids (GPLs), except for four MAbs against proteins of serotype 19. The serotype- and MAIS complex-specific epitopes on GPLs are exposed on the mycobacterial cell wall, in contrast with the common mycobacterial glycolipid, which is probably located inside the cell wall. The serotype-specific MAbs reacted with native as well as deacetylated GPLs, in contrast with the MAIS complex-specific MAbs, which reacted only with native GPLs. The MAbs will be useful for the identification of MAIS complex and M. avium serotypes 2 and 4 and a strain of serotype 19, GPL analyses with immuno-thin-layer chromatography, and the localization of GPL epitopes in mycobacteria.

摘要

制备了血清型特异性以及鸟分枝杆菌-胞内分枝杆菌-瘰疬分枝杆菌复合群(MAIS复合群)特异性单克隆抗体(MAb)。获得了一系列单克隆抗体,其中5种对血清型2具有特异性,3种对血清型4具有特异性,8种针对血清型19的一株菌株,2种对MAIS复合群具有特异性,还有2种针对迄今为止所测试的所有分枝杆菌共有的一种糖脂。血清型特异性和MAIS复合群特异性单克隆抗体在免疫荧光中与完整的分枝杆菌发生反应,在酶联免疫吸附测定和免疫薄层色谱中与分枝杆菌的脂质提取物发生反应。针对一种常见分枝杆菌糖脂的两种单克隆抗体仅在脂质酶联免疫吸附测定和免疫薄层色谱中发生反应。除了4种针对血清型19蛋白质的单克隆抗体外,所有单克隆抗体均针对糖肽脂(GPL)。与可能位于细胞壁内部的常见分枝杆菌糖脂相反,GPL上的血清型和MAIS复合群特异性表位暴露于分枝杆菌细胞壁上。血清型特异性单克隆抗体与天然以及脱乙酰化的GPL发生反应,而MAIS复合群特异性单克隆抗体仅与天然GPL发生反应。这些单克隆抗体将有助于MAIS复合群以及鸟分枝杆菌血清型2和4以及血清型19的一株菌株的鉴定、免疫薄层色谱法进行GPL分析以及GPL表位在分枝杆菌中的定位。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3241/313479/017bdb724d6d/iai00068-0264-a.jpg

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