Madathiparambil Madanan G, Kaleysa Kavitharaj N, Raghavan Kaleysaraj
Department of Biochemistry, University of Kerala, Kariavattom, Thiruvananthapuram 695 581, India.
Parasitol Res. 2009 Oct;105(4):1099-104. doi: 10.1007/s00436-009-1525-7. Epub 2009 Jul 9.
The excretory/secretory (ES) materials from filarial parasites form an important tool for immunodiagnosis of filariasis. We have raised monoclonal antibodies against ES proteins isolated from the medium incubated with live adult bovine filarial parasite Setaria digitata. The hybridoma were cloned and characterised with respect to the individual proteins of the ES materials. A secretory glycoprotein with molecular weight 200-kDa (gp200) was purified, localised and characterised using the specific monoclonal antibodies raised against it. The immunolocalisation study clearly showed that the protein is secreted out through the pores on the surface of both male and female parasites. The gp200 on reduced sodium dodecyl sulphate-polyacrylamide gel electrophoresis showed dissociation into 11- to 66-kDa 'ladder' polypeptides, and all of them could be identified with the same monoclonal antibody. The gp200 is normally shed along with the ES materials. The screening of human sera samples using the monoclonal antibodies give promising results which showed that gp200 could be a potent candidate protein for the immunodiagnosis of filariasis by the method of antigen detection.
丝虫寄生虫的排泄/分泌(ES)物质是丝虫病免疫诊断的重要工具。我们制备了针对从与活的成年牛丝虫寄生物指状腹腔丝虫一起孵育的培养基中分离出的ES蛋白的单克隆抗体。对杂交瘤进行克隆,并针对ES物质的各个蛋白质进行表征。一种分子量为200 kDa的分泌性糖蛋白(gp200)被纯化、定位并使用针对它制备的特异性单克隆抗体进行表征。免疫定位研究清楚地表明,该蛋白质通过雄性和雌性寄生虫表面的孔分泌出来。在还原十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上,gp200显示解离成11至66 kDa的“阶梯状”多肽,并且所有这些多肽都可以用同一单克隆抗体鉴定。gp200通常与ES物质一起脱落。使用单克隆抗体对人血清样本进行筛查取得了有希望的结果,表明gp200可能是通过抗原检测方法进行丝虫病免疫诊断的有力候选蛋白。