Preparation, production and functional characterization of anti cAMP and cGMP antibodies

Abstract

The present work was intended to evaluate the preparation of antigens, as well as the production and characterization of anti cAMP and cGMP antibodies. Such antibodies were obtained from rabbits, and we used 2'O-succinyl cyclic nucleotide derivate, conjugated with human serum albumin, as antigen. The characterization of the antibodies was monitored by their immunoreactivity with the labelled antigen [125I]-cyclic nucleotide. This assay consists in a competition between a labelled and an unlabelled antigen for a fixed number of binding sites present in the specific antibody. The antibodies were specific for the inducing antigens. Cross-reactivity tests showed low degree competition between the immunogen and other antigens. The very high affinity, high quality and specificity of the generated antibodies indicate that they may be used not only in radioimmunoassay and immunocytochemistry methodologies, but also as bioblockers of physiological pathways.