Development of a sphingosylphosphorylcholine detection system using RNA aptamers

利用 RNA 适体开发鞘氨醇磷酸胆碱检测系统

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作者:Katsunori Horii, Kazuya Omi, Yoshihito Yoshida, Yuka Imai, Nobuya Sakai, Asako Oka, Hiromi Masuda, Makio Furuichi, Tetsuji Tanimoto, Iwao Waga

Abstract

Sphingosylphosphorylcholine (SPC) is a lysosphingolipid that exerts multiple functions, including acting as a spasmogen, as a mitogenic factor for various types of cells, and sometimes as an inflammatory mediator. Currently, liquid chromatography/tandem mass spectrometry (LC/MS/MS) is used for the quantitation of SPC. However, because of the complicated procedures required it may not be cost effective, hampering its regular usage in a routine practical SPC monitoring. In this report, we have generated RNA aptamers that bind to SPC with high affinity using an in vitro selection procedure and developed an enzyme-linked aptamer assay system using the minimized SPC aptamer that can successfully distinguish SPC from the structurally related sphingosine 1-phosphate (S1P). This is the first case of the Systematic Evolution of Ligands by EXponential enrichment (SELEX) process being performed with a lysosphingolipid. The SPC aptamers would be valuable tools for the development of aptamer-based medical diagnosis and for elucidating the biological role of SPC.

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