Development of a sphingosylphosphorylcholine detection system using RNA aptamers.
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.
Horii
Katsunori
K
VALWAY Technology Center, NEC Soft, Ltd., Tokyo 136-8627, Japan. horii-katsunori@mxb.nes.nec.co.jp
Omi
Kazuya
K
Yoshida
Yoshihito
Y
Imai
Yuka
Y
Sakai
Nobuya
N
Oka
Asako
A
Masuda
Hiromi
H
Furuichi
Makio
M
Tanimoto
Tetsuji
T
Waga
Iwao
I
D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000235:genetics; D001483:Base Sequence; D001681:Biological Assay; D007700:Kinetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry; D015203:Reproducibility of Results; D052156:SELEX Aptamer Technique / Q000379:methods*; D013110:Sphingosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry; D020349:Surface Plasmon Resonance
D052157:Aptamers, Nucleotide; C005356:sphingosine phosphorylcholine; D010767:Phosphorylcholine; D013110:Sphingosine