Preparation of polyamine antibody and its use in enzyme immunoassay of spermine and spermidine with beta-D-galactosidase as a label.
An enzyme immunoassay for polyamines is described which uses beta-galactosidase labeled spermine and antiserum raised in rabbits against spermine-bovine serum albumin synthesized by coupling spermine to mercaptosuccinylated bovine serum albumin with a bifunctional cross-linker, N-(gamma-maleimidobutyryloxy)-succinimide. The lower limit of detection by this assay, which involves a double antibody technique for the separation of antibody-bound and free antigen, was 1 ng of spermine per tube. The anti-spermine serum showed 88% cross-reaction with spermidine but only 0.13% with putrescine, 0.08% with 1,3-diaminopropane, and 0.04% with cadaverine. The method has been used to measure serum polyamine levels in healthy subjects and cancer patients, in whom mean concentrations of 58.1 ng/ml and 94.8 ng/ml (as spermine), were respectively noted. This enzyme immunoassay is specific, accurate and easy to perform, and appears suitable for routine clinical use.
D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D005696:Galactosidases / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D009369:Neoplasms / Q000032:analysis; D011817:Rabbits; D013095:Spermidine / Q000097:blood*; D013096:Spermine / Q000097:blood* / Q000276:immunology
D013096:Spermine; D005696:Galactosidases; D013095:Spermidine