PRODUCTION OF MONOCLONAL ANTIBODIES AGAINST TOXOPLASMA GONDII
WEE LYE KENG JANET
WEE LYE KENG JANET
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Abstract
Several hybridomas producing antibodies detected by the enzyme-linked imnunosorbent assay {ELISA) were established in fusions involving mouse myeloma P3. X63. g8.Ul and spleen cells from BALB/c mice immunized against the Rh strain of Toxoplasma gondii. Three types of immunization routes were attempted --
(1) in vivo with formalin-killed tachyzoites, (2) in vivo with soluble antigen extract, and (3) in vitro with soluble antigen extract. Six strongly positive hybrids (by ELISA) were selected for cloning and characterization. From the first route of immunization, hybrids 4G4 and 1G7 established were determined to secrete antibodies of subclasses IgG3 and IgG2a respectively. Hybrids 2E5, 3F2 and 5F3 from the second route were found to be IgGl producers. 0nly hybrid 2B8 (from the third route) secreted IgM antibodies. By the Western blot analysis, antibodies of the hybrids induced by in vivo immunization reacted with one to six antigenic determinants whose apparent molecular weights ranged from 14.7 - 20.8 kd. Hybrid 2B8 (IgM secreting) reacted with a wider range of determinants; from 14 kd to those with molecular weights greater than 94.1 kd.
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Date
1986
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