Antipolyamine antibodies were produced following immunization of New Zealand white rabbits with thyroglobulin spermine conjugate, using multiple-site injection and macrophage-harvesting techniques. The antispermine antibody in a radioimmunoassay system cross-reacted with spermidine (22%), putrescine (1%), diaminopropane (6%), and cadaverine (0.16%). No reaction with L-lysine, ornithine, or histamine was noted. Using this antibody, the radioimmunoassay for direct determination of serum polyamines was developed. A sensitivity of 200 pg was achieved on human serum samples. This radioimmunoassay would appear to be a useful tool for the detection and monitoring of a variety of pathophysiological states associated with abnormal polyamine metabolism, i.e., cancer.