Horn C, Pescher P, Romain F, Marchal G
Unité de Physiopathologie de l'Infection, Institut Pasteur, Paris, France.
J Immunol Methods. 1996 Oct 16;197(1-2):151-9. doi: 10.1016/0022-1759(96)00141-x.
The alanine-proline antigen (APA), representing less than 2% of the released or excreted material during Mycobacterium tuberculosis or bacillus Calmette-Guérin (BCG) growth, is a dominant antigen present during M. tuberculosis infection or BCG immunization. To obtain new tools to dissect the major epitopes of the APA molecules, seven monoclonal antibodies (mAbs) against the purified molecules were developed. Epitope maps of the mAbs were obtained on APA molecules absorbed on plastic surfaces or in solution (BIAcore technology). The mAbs were found to be independent or to be different despite binding to adjacent or overlapping epitopes. In Western blot assays some proteins secreted in culture fluid by M. avium, M. kansasii, M. smegmatis or M. xenopi were also labelled by six of the seven antibodies. Conversely one antibody was specific for the proteins from the M. tuberculosis complex (I10-0,3) demonstrating that the APA molecules have some properties or general conformations that are specific for M. tuberculosis and M. bovis.
丙氨酸 - 脯氨酸抗原（APA）在结核分枝杆菌或卡介苗（BCG）生长过程中释放或排泄的物质中占比不到2%，是结核分枝杆菌感染或卡介苗免疫期间存在的主要抗原。为了获得剖析APA分子主要表位的新工具，研发了七种针对纯化分子的单克隆抗体（mAb）。通过BIAcore技术在吸附于塑料表面或溶液中的APA分子上获得了这些mAb的表位图谱。尽管这些mAb结合相邻或重叠表位，但发现它们相互独立或不同。在蛋白质印迹分析中，七种抗体中的六种也标记了鸟分枝杆菌、堪萨斯分枝杆菌、耻垢分枝杆菌或偶发分枝杆菌在培养液中分泌的一些蛋白质。相反，一种抗体对结核分枝杆菌复合群（I10 - 0,3）的蛋白质具有特异性，这表明APA分子具有一些对结核分枝杆菌和牛分枝杆菌特异的特性或一般构象。
