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HYBRIDOMA PRODUCTION USING IMMUNE LYMPHOCYTES AGAINST BRUGIA MALAYI ANTIGEN WITH MYELOMA CELLS | Soeyoko | Buletin Penelitian Kesehatan

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  • HYBRIDOMA PRODUCTION USING IMMUNE LYMPHOCYTES AGAINST BRUGIA MALAYI ANTIGEN WITH MYELOMA CELLS

    Soeyoko Soeyoko, Noerhajati S., Abdul Salam Sofro

    Abstract


    Wuchereria bancrofti, Brugia malayi, and Brugia timori are the causative agents of lymphatic hiariasis in Indonesia, but in some endemic areas, Brugia malayi is the most commonly found,Diagnosis of hiariasis is normally based on clinical and parasitological examinations, but both have limitations. Therefore now the immunological examination plays an important role in the diagnosis of filariasis. The discovery of monoclonal antibodies recently may probably give a firm scientific basis in immunology and add a new dimension to the efforts of developing a specific and sensitive immunological test for various stages of filarial infection.

    In this study, the production of hybridoma cells to develop monoclonal antibodies against B. malayi integrated a number of techniques: preparations of B. malayi surface antigen, immune lymphocyte cells, NSI myeloma cells and macrophage feeder layers, and a fusion of immune lymphocytes with myeloma cells.

    The results of this study can be concluded in three points:

    1. Protein analysis of the surface antigen was examined by Biureet and SDS-PAGE. A total of fourteen examinations were conducted by using 4000 L3 for each experiment. Three were not detected by Biureet method, but showed five protein fractions by SDS-PAGE. The protein concentration of the surface L3 was varied from 85.0/µg/ml to 769.23/µg/ml, with an average of 297.04/µg/ml.
    2. The immunoreactivifies of Balb/c mice antibodies to B. malayi L3 surface antigen were tested by ELISA and showed a gradual increase after four times immunizations at two weeks interval. The optical density (OD) after four times immunizations was varied from 0.363 to 0.878 each mouse, where as the positive control sera OD was 0.570.
    3. Hybridization using immune lymphocytes against B. malayi L3 surface antigen with myeloma cells yielded 60.41% hybrid cells and none of them produced monoclonal antibodies tested of ELISA.

    Keywords


    Brugia Malayi; Myeloma Cell; Hybridoma; Immune Lymphocytes

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    Buletin Penelitian Kesehatan (Bulletin of Health Research, ISSN: 0125-9695. EISSN: 2338-3453) diterbitkan oleh National Institute of Health Research and Development, Ministry of Health of Republic of Indonesia
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