Shin Sung Jae, Lee Seung-Sub, Manning Elizabeth J B, Collins Michael T
Department of Microbiology and Infection Signaling Network Research Center, Research Institute for Medical Sciences, College of Medicine, Chungnam National University, Daejeon, Republic of Korea.
J Microbiol. 2009 Oct;47(5):600-9. doi: 10.1007/s12275-009-0052-7. Epub 2009 Oct 24.
Antibodies specific to the cell surface antigens of Mycobacterium avium subsp. paratuberculosis (MAP) have multiple useful applications, e.g. organism detection, immunoconcentration, and cell visualization. The aim of this study was to produce and compare polyclonal antibodies for such research and diagnostic purposes. Three polyclonal antibodies to MAP were produced using sera from immunized rabbits and chickens plus naturally infected cows. Cross-reactive antibodies in each MAP antibody preparation were removed by absorption with heterologous mycobacterial and non-mycobacterial cells. The specificity of each resulting polyclonal antibody preparation was evaluated by ELISA to multiple bacterial cell wall extract antigens. After absorption, chicken anti-MAP IgY had the highest specificity of the three antibody preparations. FITC-la-beled anti-MAP IgY was used to effectively locate MAP in macrophages 12 h post-infection. Also, immunomagnetic beads coated with anti-MAP IgY enhanced recovery of MAP from bacterial suspensions in comparison with non-antibody coated beads. Anti-MAP IgY provides a novel new reagent with broad diagnostic and research applications requiring specific concentration, detection, and quantification of MAP.
针对副结核分枝杆菌（MAP）细胞表面抗原的特异性抗体具有多种有用的应用，例如微生物检测、免疫富集和细胞可视化。本研究的目的是为此类研究和诊断目的制备并比较多克隆抗体。使用免疫兔和鸡以及自然感染牛的血清制备了三种针对MAP的多克隆抗体。每种MAP抗体制剂中的交叉反应性抗体通过用异源分枝杆菌和非分枝杆菌细胞吸收而去除。通过ELISA对多种细菌细胞壁提取物抗原评估每种所得多克隆抗体制剂的特异性。吸收后，鸡抗MAP IgY在三种抗体制剂中具有最高的特异性。用异硫氰酸荧光素（FITC）标记的抗MAP IgY用于在感染后12小时有效定位巨噬细胞中的MAP。此外，与未包被抗体的磁珠相比，包被抗MAP IgY的免疫磁珠提高了从细菌悬液中回收MAP的效率。抗MAP IgY提供了一种新型试剂，具有广泛的诊断和研究应用，可用于MAP的特异性富集、检测和定量。
