Production of Monoclonal Antibodies for Toxoplasma gondii Lactate Dehydrogenase

Abstract

"Background/Purpose: Toxoplasma gondii is one of the most common parasitic infections of human and other warm blooded animals. Because of the geographical distribution of toxoplasmosis, there is a need to develop a sensitive, rapid, simple and an inexpensive test for the early screening of patiens. In this study, to create the dipstick kit that has high sensitivity against LDH (lactate dehydrogenase) of T. gondii, at the first of all, hybridoma cell lines producing monoclonal antibodies with high affinity and specificity against T. gondii LDH (TgLDH) were established.

Methods: After 6-8 week old BALB/c mice were immunized with synthetic peptides of TgLDH, splenocytes were harvested. To get the monoclonal antibodies (McAbs), splenocytes were fused with myeloma cells using PEG by a cell fusion technique. Hybridoma cells were selected by ELISA and subcloned through a limiting dilution method. McAbs were characterized by isotyping.

Results: Eight McAb-producing hybridoma cells were selected. After three times of limiting dilution, eight clones, 1A10, 1B12, 3B2, 3B6, 1C12, 3C8, 3G6 and 3H1, were selected. OD values of eight monoclonal antibodies were ranged from 0.938 to 1.269 by ELISA. All eight McAbs isotypes were IgM class.

Conclusion: These results indicated that eight hybridoma clones had strong reactivity to TgLDH and may be useful for diagnosis to T. gondii infection.

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