Subrahmanyam D, Parab P B, Rajasekariah G R, Rudin W, Betschart B, Weiss N
Hindustan Ciba-Geigy Research Centre, Bombay, India.
Acta Trop. 1990 Jul;47(5-6):381-90. doi: 10.1016/0001-706x(90)90039-3.
Monoclonal antibodies (mAbs) have been prepared against excretory-secretory-metabolic (ESM) antigens of microfilariae (mf) of Wuchereria bancrofti (WbmfESM) and against third stage larvae (L3) of Brugia malayi (BmL3), and purified from ascites fluids with ammonium sulphate. Both antibodies were of the IgM type and did not react with phosphorycholine. The mAb against BmL3 (F46) reacted in ELISA with antigens of L3 of B. malayi, B. pahangi and W. bancrofti and of adults of B. malayi. The mAb raised against wbmfESM (F32) resembled F46 in this respect, though with a lower titer towards the antigens, and in addition reacted with the ESM-antigens of mf and of L3 of W. bancrofti. F46 was able to detect L3 antigens of filarial parasites in spiked serum samples with a detection limit of 8-16 ng in absolute amount. The antibody was found to label the cuticular portion of L3 and adults of the lymphatic parasites, and not the epicuticular surface, in immunoelectron microscopic studies. The antibody recognized a 36 kDa component of the beta-mercaptoethanol extracts of B. pahangi-adults in Western blot analysis.
已制备出针对班氏吴策线虫微丝蚴(Wbmf)排泄分泌代谢(ESM)抗原以及马来布鲁线虫三期幼虫(BmL3)的单克隆抗体(mAb),并通过硫酸铵从腹水液中进行纯化。两种抗体均为IgM型,且不与磷酸胆碱发生反应。抗BmL3的单克隆抗体(F46)在酶联免疫吸附测定(ELISA)中能与马来布鲁线虫、彭亨布鲁线虫的L3抗原以及班氏吴策线虫的L3抗原和马来布鲁线虫成虫抗原发生反应。针对Wbmf ESM产生的单克隆抗体(F32)在这方面与F46相似,不过对这些抗原的效价较低,此外还能与班氏吴策线虫微丝蚴和L3的ESM抗原发生反应。F46能够在加标的血清样本中检测到丝虫寄生虫的L3抗原,绝对检测限为8 - 16纳克。在免疫电子显微镜研究中发现,该抗体标记淋巴寄生虫L3和成虫的角质层部分,而非表皮表面。在蛋白质印迹分析中,该抗体识别彭亨布鲁线虫成虫β - 巯基乙醇提取物中的一种36 kDa成分。