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Korean J Parasitol > Volume 31(1):1993 > Article

Original Article
Korean J Parasitol. 1993 Mar;31(1):37-42. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1993.31.1.37
Copyright © 1993 by The Korean Society for Parasitology
Identification of surface antigens of Trichomonas vaginalis
Nam-Sick Woo,1Duk-Young Min,*1Mi-Hyea Leem,1 and Yong-Keel Choi2
1Department of Parasitology, College of Medicine, Hanyang University, Seoul 133-791, Korea.
2Department of Biology, College of Natural Science, Hanyang University, Seoul 133-791, Korea.
Received December 28, 1992; Accepted January 26, 1993.

Abstract

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.

Figures


Fig. 1
Immunoprecipitation of biotinylated T. vaginalis. 120 µl aliquots of trichomonal proteins labelled at concentrations of 1 mM (A) or 10 mM (B) NHS-biotin were incubated with 50 µl of normal (lane 5) and immune rabbit serum in the presence of 0 (lane 1), 125 (lane 2), 450 (lane 3) or 1075 µl (lane 4) lysis buffer respectively. Immunoprecipitated complexes were separated by SDS-PAGE and electroblotting and labelled proteins were detected by antibiotin-HRP.


Fig. 2
Western blots of biotin-labelled proteins of T. vaginalis HY-1 obtained from intact cells (lane 1) and from control cells disrupted prior to labelling (lane 2). A: Amidio black staining. B: Anti-biotin detection of biotinylated proteins. C: immuno-precipitated proteins labelled by NHS-biotin and detected by anti-biotin-HRP.


Fig. 3
Western blots of biotin-labelled proteins of T. vaginalis HY-1 (lane-1), HY-15 (lane 2) and ATCC 50148 (lane 3). A: Amido black staining of T. vaginalis proteins. B: Anti-biotin detection of biotinylated proteins obtained from intact cells. C: Immunoprecipitation of biotinylated proteins obtained from intact cells.

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