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利用 RNA 适体开发鞘氨醇磷酸胆碱检测系统。

Development of a sphingosylphosphorylcholine detection system using RNA aptamers.

机构信息

VALWAY Technology Center, NEC Soft, Ltd., Tokyo 136-8627, Japan.

出版信息

Molecules. 2010 Aug 20;15(8):5742-55. doi: 10.3390/molecules15085742.

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.

摘要

鞘氨醇磷胆碱(SPC)是一种溶血神经鞘脂,具有多种功能,包括作为痉挛原、各种类型细胞的有丝分裂原,有时还作为炎症介质。目前,液相色谱/串联质谱(LC/MS/MS)用于 SPC 的定量。然而,由于需要复杂的程序,它可能不具有成本效益,阻碍了其在常规实际 SPC 监测中的常规使用。在本报告中,我们使用体外选择程序生成了与 SPC 具有高亲和力的 RNA 适体,并开发了一种使用最小化的 SPC 适体的酶联适体测定系统,该系统能够成功区分 SPC 与结构上相关的鞘氨醇 1-磷酸(S1P)。这是首次使用溶血神经鞘脂进行系统进化配体的指数富集(SELEX)过程。SPC 适体将是开发基于适体的医学诊断和阐明 SPC 生物学作用的有价值的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/73aa/6257670/809c322cc85a/molecules-15-05742-g001.jpg

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