Guanylic-acid-specific antibodies were elicited in rabbits) using as immunogen pG linked through 5′-phosphate to thyroglobulin. Specificity and affinity of antibodies to nucleotides) nucleosides, DNA, and RNA were studied by their binding to radioactive ligands and competition experiments. Guanylic-acid-specific antibodies do not bind to deoxyguanylic acid and have an average association constant of 107 M−1 at 4°C Binding of the antibodies to3H-RNA is G-specifiC. The antibodies do not bind to32P-ssDNA or32P-dsDNA. The pG-specific antibodies could be separated into different fractions by affinity chromatography. These fractions) though specific to pG) differ in their cross-reactivities to nucleosides and nucleotides.
Abbreviations used G, guanosine; pG, guanosine 5′-phosphate (similarly for other nucleotides); dpG, deoxyriboguanosine 5′-phosphate; ssDNA, single-stranded DNA; dsDNA, double-stranded DNA; Tg, thyroglobulin; BSA, bovine serum albumin; RSA, rabbit serum albumin; EDC, 1-ethyl-1-3 (3-dimethylisopropyl) carbodiimide; Ac-Lys-NHMe, N-α-acetyl L-lysine N-methylamide; Ac-Lys-(pG)-NHMe, pG coupled through the phosphate to the ∊-amino group of Ac-Lys-NHMe; AH-Sepharose-pG, aminohexyl-Sepharose-pG; TBS, Tris-buffered saline; SLE, systemic lupus erythematosus;