PREPARATION AND CHARACTERIZATION OF POLYCLONAL AND MONOCLONAL-ANTIBODIES TO POLYAMINES

In order to develop an immunocytochemical method suitable for the study of the cellular localization and intracellular distribution of polyamines we have prepared and characterized antibodies to polyamines. Artificial immunogens were prepared by coupling putrescine, spermidine and spermine to a carrier protein. Immunogens containing bovine serum albumin as a carrier protein were used to immunize rabbits (polyclonal antibodies) and mice (for the production of Mabs). The specificity of the antibodies was tested in an ELISA system utilizing antigens synthesized from thyroglobulin and one of the polyamines. Polyclonal antibodies to putrescine, spermidine and spermine were obtained. In order to develop an immunocytochemical method suitable for the study of the cellular localization and intracellular distribution of polyamines we h ave prepared and characterized antibodies to polyamines. Artificial immuno gens were prepared by coupling putrescine, spermidine and spermine to a car rier protein. Immunogens containing bovine serum albumin as a carrier protein were used to immunize rabbits (polyclonal antibodies) and mice (for the production of Mabs). The specificity of the antibodies was tested in a n ELISA system utilizing antigens synthesized from thryoglobulin and one of the pollyamines. Polyclonal antibodies to putrescine, spermidine and sperm ine were obtained. However, these antibodies showed a variable degree of cross-reactivity to the polyamines not used for immunization. Two hybridom a cell lines were developed. The first, MPut88, selectively produces a Mab to putrescine, the second, MSpm/d88 produces a Mab which recognizes spermi ne and spermidine but does not react with putrescine.