Arora S K, Sehgal S
Department of Immunopathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India.
Med Microbiol Immunol. 1989;178(2):81-8. doi: 10.1007/BF00203303.
The study was conducted with the prime objective of isolating an antigen from the crude preparation of whole promastigotes (Leishmania donovani) with a view to future exploitation in serodiagnosis and production of monoclonal antibodies. Soluble antigen, prepared from promastigotes isolated from actively growing cultures, was fractionated by gel filtration chromatography over a column of Sephadex G200. Three peaks of proteins could be recovered. The antigenic reactivity of different fractions was checked against sera of kala azar patients by immunoelectrophoresis (IEP), rocket immunoelectrophoresis (RIEP) and enzyme-linked immunosorbent assay (ELISA). The first peak was found to be highly reactive in comparison with other peaks. SDS-PAGE and western blot analysis of this antigen revealed a major antigen of promastigotes in the vicinity of 65 to 66 kDa, while a weakly reactive triplet could also be detected in the low molecular weight region.
本研究的主要目的是从杜氏利什曼原虫前鞭毛体的粗制品中分离出一种抗原，以便将来用于血清诊断和单克隆抗体的生产。从活跃生长的培养物中分离出的前鞭毛体制备的可溶性抗原，通过在Sephadex G200柱上进行凝胶过滤色谱法进行分级分离。可以回收三个蛋白质峰。通过免疫电泳（IEP）、火箭免疫电泳（RIEP）和酶联免疫吸附测定（ELISA）检测不同组分与黑热病患者血清的抗原反应性。发现第一个峰与其他峰相比具有高反应性。对该抗原的SDS-PAGE和蛋白质印迹分析显示，在65至66 kDa附近有前鞭毛体的主要抗原，而在低分子量区域也可检测到弱反应性三联体。
