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Tsutsumi, T. et al.

Anti-idiotype monoclonal antibodies against anti-microcystin antibody and their use in enzyme immunoassay.

Microcystins (MCs), a group of heptapeptide hepatotoxins produced by cyanobacteria, are suspected as tumor-promoter contaminants of environmental water. We have previously developed an enzyme-linked immunosorbent assay (ELISA) for MCs based on an anti-MC MAb (MAb-mc). We describe here the production of anti-idiotype MAbs (MAb-ids) which react with MAb-mc and their use in a new ELISA for MCs. For the production of MAb-id, hybridoma cells were generated from mice immunized with MAb-mc. Two MAbs were selected for their ability to inhibit the binding of MAb-mc to microcystin-LR (MCLR)-bovine serum albumin conjugate in ELISA. The one with the higher inhibitory activity, designated Id7 ( IgG1, kappa), was further characterized. ELISA and immunoprecipitation analysis revealed that Id7 specifically bound to MAb-mc but not to control IgG1, and the binding was inhibited by free MCLR. Therefore, Id7 is a MAb-id to MAb-mc and potentially possesses the structural image of MCLR. To establish MAb-id based ELISA, Id7 was tested for use in three types of competitive ELISA for MCs. The best format enabled reliable measurements of MCLR in the range of 100-1000 pg/ml with a coefficient of variation of less than 3%. In addition, microcystin-RR and microcystin-YR, principal MCs found in environmental water, were cross-reacted well (67-111% of MCLR) in the ELISA although 6(Z)-MCLR, a minor component, was less reactive (7% of MCLR). A comparative study of the MAb-id based ELISA with previously established MAb-mc based ELISA revealed good correlation (n = 14, r = 0.97) between the two methods for measurements of MCs content in freshwater samples. Thus, developed MAb-id based ELISA is an useful alternative for environmental monitoring of MCs.[1]

References

Anti-idiotype monoclonal antibodies against anti-microcystin antibody and their use in enzyme immunoassay. Tsutsumi, T., Nagata, S., Yoshida, F., Ueno, Y. Toxicon (1998) [Pubmed]

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