Rapid purification of iodinated ligands for cyclic nucleotide radioimmunoassays.
The tyrosine methyl esters of succinyl cyclic AMP and succinyl cyclic GMP were iodinated by the chloramine T method and individually applied to C18 cartridges. A solution of 1-propanol/0.1 M sodium acetate pH 4.75 (17.5:82.5) was then pumped onto each cartridge and the eluate collected. A large peak of radioactivity, containing primarily the monoiodo and diiodo derivatives, was eluted. Radioactivity in peak fractions was greater than or equal to 95% the monoiodo derivative and represented 20 to 25% of the starting radioactivity. Contamination by the native cyclic nucleotide analogs was less than 5%. These peak fractions containing primarily monoiodinated products worked well in cyclic nucleotide radioimmunoassays. This fractionation required less than 30 min.
Wilson
S P
SP
Department of Pharmacology, University of South Carolina School of Medicine, Columbia 29208.
D002851:Chromatography, High Pressure Liquid; D000242:Cyclic AMP / Q000032:analysis*; D006152:Cyclic GMP / Q000032:analysis*; D007457:Iodine Radioisotopes; D008024:Ligands / Q000302:isolation & purification*; D011863:Radioimmunoassay
D007457:Iodine Radioisotopes; D008024:Ligands; D000242:Cyclic AMP; D006152:Cyclic GMP