Preparation, characterization, and use of an antiserum specific for 2',5'-oligoadenylates

Abstract Periodate-oxidized ppp(A2'p)3A was coupled in high yield to an acyl hydrazide derivative of bovine serum albumin and the conjugate was used to immunize rabbits. Several potent and specific antisera were obtained, and one was tested extensively. Fifty per cent of the bound probe ppp(A2'p)3A3'[32P]pCp was displaced by compounds containing the moiety -pA2'pA-at concentrations of 20-60 nM and by compounds containing the moiety -p(A2'p)2A- at 1 nM. The 3',5'-oligoadenylate (A3'p)3A was bound more than 10,000 times less tightly than (A2'p)3A. The antiserum can be used in competition assays in which the amount of an antibody-probe complex is measured after binding to Millipore filters or to polystyrene beads. These assays allow the clear detection of as little as 20 fmol (20 microliters of 1 nM) of 2',5'-oligoadenylates.