Woo N S, Min D Y, Leem M H, Choi Y K
Department of Parasitology, College of Medicine, Hanyang University, Seoul, Korea.
Korean J Parasitol. 1993 Mar;31(1):37-42. doi: 10.3347/kjp.1993.31.1.37.
Surface proteins of Trichomonas vaginalis (T. vaginalis) were analyzed to study the antigenic variation. The surface proteins of protozoa were labelled by N-hydroxysuccinimide-biotin (NHS-biotin), the NHS-biotin-labelled proteins were immunoprecipitated with rabbit antiserum to purify the antigenic fractions and analysed by SDS-PAGE plus electroblotting. The results obtained in this study were as follows; Biotinylated T. vaginalis-proteins obtained from intact cell and cells disrupted prior to labelling were detected by anti-biotin-peroxidase in Western blots. Labelled proteins were immunoprecipitated by T. vaginalis-immunized rabbit serum and the six bands with the molecular weights of 46, 60, 68, 90, 130 and 220 kDa were identified as having antigenicity. T. vaginalis HY-1, HY-15 and ATCC 50148 were immunoprecipitated by immune rabbit serum after biotinylation and there were no difference from antigenic bands among these strains by this technique. In conclusion with the results obtained in the present study, it was assumed that surface proteins of T. vaginalis were labelled by biotinylation and the six labelled bands at 46, 60, 68, 90, 130 and 220 kDa in their molecular weight were identified as having antigenicity by immunoprecipitation (IP) and this biotinylation-IP technique may be used for further study of surface antigen of T. vaginalis.
对阴道毛滴虫的表面蛋白进行分析以研究抗原变异。用N-羟基琥珀酰亚胺-生物素(NHS-生物素)标记原生动物的表面蛋白,将NHS-生物素标记的蛋白用兔抗血清进行免疫沉淀以纯化抗原组分,并通过SDS-PAGE加电印迹法进行分析。本研究获得的结果如下:在Western印迹中用抗生物素-过氧化物酶检测从完整细胞以及在标记前被破坏的细胞中获得的生物素化阴道毛滴虫蛋白。标记的蛋白被阴道毛滴虫免疫的兔血清免疫沉淀,并且鉴定出分子量为46、60、68、90、130和220 kDa的六条带具有抗原性。生物素化后,阴道毛滴虫HY-1、HY-15和ATCC 50148被免疫兔血清免疫沉淀,通过该技术在这些菌株的抗原条带之间没有差异。根据本研究获得的结果得出结论,推测阴道毛滴虫的表面蛋白通过生物素化进行标记,并且通过免疫沉淀(IP)鉴定出分子量为46、60、68、90、130和220 kDa的六条标记带具有抗原性,并且这种生物素化-IP技术可用于进一步研究阴道毛滴虫的表面抗原。