用于环磷酸鸟苷的阿托摩尔灵敏放射免疫测定法。

Attomole sensitive radioimmunoassay for cyclic GMP.

作者信息

Axelsson K L, Bornefeldt K E, Norlander B, Wikberg J E

机构信息

Department of Pharmacology, Linköping University, Sweden.

出版信息

Second Messengers Phosphoproteins. 1988;12(4):145-54.

PMID:2854558

Abstract

The sensitivity of the radioimmunoassay for cyclic GMP has been improved to readily detect attomole (200-300) amounts of the nucleotide in tissue extracts. The improved sensitivity has been achieved by using a high specific radioactivity [125I] cyclic GMP 2'O-succinyl tyrosine methyl ester (2200 Ci/mmole) as labeled antigen in conjunction with acetylation of the cyclic GMP in the samples as well as by using a selected cyclic GMP antibody. The high specific [125I] cyclic GMP 2'O-succinyl tyrosine methyl ester was prepared radiochemically pure by radioiodination of cyclic GMP 2'O-succinyl tyrosine methyl ester and purification using HPLC. The new attomole sensitive cyclic GMP radioimmunoassay is simple to perform and fast and has been applied with success to determine attomole quantities of cyclic GMP in cultured smooth muscle cells and AG 1523 fibroblasts.

摘要

用于环磷酸鸟苷（cGMP）的放射免疫分析的灵敏度已得到提高，能够轻松检测组织提取物中阿托摩尔（200 - 300）量的该核苷酸。通过使用高比放射性的[125I]环磷酸鸟苷2'-琥珀酰酪氨酸甲酯（2200 Ci/mmol）作为标记抗原，并结合对样品中环磷酸鸟苷进行乙酰化处理，以及使用选定的环磷酸鸟苷抗体，实现了灵敏度的提高。高比放射性的[125I]环磷酸鸟苷2'-琥珀酰酪氨酸甲酯通过对环磷酸鸟苷2'-琥珀酰酪氨酸甲酯进行放射性碘化并使用高效液相色谱法（HPLC）纯化而制备成放射化学纯品。新型阿托摩尔灵敏的环磷酸鸟苷放射免疫分析操作简单、快速，已成功应用于测定培养的平滑肌细胞和AG 1523成纤维细胞中阿托摩尔量的环磷酸鸟苷。