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Development of a Sphingosylphosphorylcholine Detection System Using RNA Aptamers
VALWAY Technology Center, NEC Soft, Ltd., Tokyo 136-8627, Japan
Biotechnology Research Group, Fundamental Research Department, Fujirebio Inc., Tokyo, 192-0031, Japan
* Author to whom correspondence should be addressed.
Received: 27 July 2010; in revised form: 16 August 2010 / Accepted: 17 August 2010 / Published: 20 August 2010
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.
Keywords: aptamer; enzyme-linked aptamer assay; sphingosine 1-phosphate; sphingosylphosphorylcholine; systematic evolution of ligands by exponential enrichment
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Horii, K.; Omi, K.; Yoshida, Y.; Imai, Y.; Sakai, N.; Oka, A.; Masuda, H.; Furuichi, M.; Tanimoto, T.; Waga, I. Development of a Sphingosylphosphorylcholine Detection System Using RNA Aptamers. Molecules 2010, 15, 5742-5755.
Horii K, Omi K, Yoshida Y, Imai Y, Sakai N, Oka A, Masuda H, Furuichi M, Tanimoto T, Waga I. Development of a Sphingosylphosphorylcholine Detection System Using RNA Aptamers. Molecules. 2010; 15(8):5742-5755.
Horii, Katsunori; Omi, Kazuya; Yoshida, Yoshihito; Imai, Yuka; Sakai, Nobuya; Oka, Asako; Masuda, Hiromi; Furuichi, Makio; Tanimoto, Tetsuji; Waga, Iwao. 2010. "Development of a Sphingosylphosphorylcholine Detection System Using RNA Aptamers." Molecules 15, no. 8: 5742-5755.