Sinha A K, Colman R W
Eur J Biochem. 1977 Mar 1;73(2):367-71. doi: 10.1111/j.1432-1033.1977.tb11327.x.
Using the principle of affinity elution chromatography, a microassay has been developed for the determination of picomolar and even smaller quantities of adenosine 3'.5' -monophosphate (cyclic AMP) in biological samples. In principle, cyclic AMP along with the other anions is adsorbed onto an anion exchange cellulose and subsequently, the cyclic nucleotide is specifically eluted with cyclic-AMP -dependent protein kinase. In the actual assay cyclic AMP is determined by competitive displacement of radiolabelled cyclic AMP by non-labelled cyclic nucleotide. The described procedure results in a linear dependence for the displacement of the radioactive nucleotide on cyclic AMP concentration and obviates the usual procedure of sample enrichment by lyophilization. The degree of interference of different nucleotides including cyclic GMP was found to be minimal and in this respect the use affinity elution chromatography was comparable to radioimmunoassay.
利用亲和洗脱色谱原理，已开发出一种微量测定法，用于测定生物样品中皮摩尔甚至更小量的3',5'-环磷酸腺苷（环磷酸腺苷）。原则上，环磷酸腺苷与其他阴离子一起吸附到阴离子交换纤维素上，随后，环核苷酸用依赖环磷酸腺苷的蛋白激酶特异性洗脱。在实际测定中，环磷酸腺苷是通过未标记的环核苷酸竞争性取代放射性标记的环磷酸腺苷来测定的。所描述的方法导致放射性核苷酸的取代与环磷酸腺苷浓度呈线性关系，并且避免了通常通过冻干进行样品富集的过程。发现包括环鸟苷酸在内的不同核苷酸的干扰程度最小，在这方面，亲和洗脱色谱的使用与放射免疫测定相当。
