中国科学院大连化学物理研究所机构知识库
Advanced  
DICP OpenIR  > 中国科学院大连化学物理研究所  > 期刊论文
题名: Chemiluminescently labeled aptamers as the affinity probe for interaction analysis by capillary electrophoresis
作者: Li, Hong-Yi1;  Deng, Qin-Pei1;  Zhang, De-Wen1;  Zhou, Ying-Lin1;  Zhang, Xin-Xiang1
关键词: Affinity probe ;  Aptamer ;  CE ;  Chemiluminescence ;  Interaction analysis
刊名: ELECTROPHORESIS
发表日期: 2010-07-01
DOI: 10.1002/elps.201000131
卷: 31, 期:14, 页:2452-2460
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: Aptamers are nucleic acid oligonucleotides, which can recognize targets with high affinity and specificity. Fluorescently labeled aptamers have been used as affinity probes in CE for interaction analysis. In this study, a method of labeling aptamers chemiluminescently with isoluminol isothiocyanate (ILITC) through covalent bonds was proposed and realized. The ILITC-labeled aptamers were characterized by HPLC-MS and purified by HPLC. After desalination, the ILITC-labeled aptamers were employed as the affinity probe for interaction analysis in CE coupled with chemiluminescence detection (CE-CL) by interface of end column reaction mode, the apparatus of which was home-designed and setup. CE-CL experiment conditions, including buffer pH, concentrations of horseradish peroxidase and H(2)O(2), were optimized first. The system of thrombin and its 29-mer aptamer was chosen as the model. Binding parameters, namely the dissociation constant (K(d)) and the binding site number (n), were calculated. The K(d) obtained was 124.0 +/- 6.9 nM in agreement with the reported values. Thus, interaction analysis method based on chemiluminescently labeled aptamers as the affinity probe in CE-CL has been established. This method can be widely applied due to the ease and universality of the labeling method, simplicity of CE-CL apparatus and combination with aptamers for a wide range of targets.
关键词[WOS]: IN-VITRO SELECTION ;  HUMAN THROMBIN ;  IMMUNOASSAY ;  CELL ;  RNA ;  SIMPLIFICATION ;  SENSITIZATION ;  COMPLEXES ;  LIGANDS ;  SELEX
语种: 英语
WOS记录号: WOS:000280709700021
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/141729
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

Files in This Item:

There are no files associated with this item.


作者单位: 1.Peking Univ, BNLMS, Key Lab Bioorgan Chem & Mol Engn, Minist Educ,Inst Analyt Chem,Coll Chem, Beijing 100871, Peoples R China

Recommended Citation:
Li, Hong-Yi,Deng, Qin-Pei,Zhang, De-Wen,et al. Chemiluminescently labeled aptamers as the affinity probe for interaction analysis by capillary electrophoresis[J]. ELECTROPHORESIS,2010,31(14):2452-2460.
Service
 Recommend this item
 Sava as my favorate item
 Show this item's statistics
 Export Endnote File
Google Scholar
 Similar articles in Google Scholar
 [Li, Hong-Yi]'s Articles
 [Deng, Qin-Pei]'s Articles
 [Zhang, De-Wen]'s Articles
CSDL cross search
 Similar articles in CSDL Cross Search
 [Li, Hong-Yi]‘s Articles
 [Deng, Qin-Pei]‘s Articles
 [Zhang, De-Wen]‘s Articles
Related Copyright Policies
Null
Social Bookmarking
  Add to CiteULike  Add to Connotea  Add to Del.icio.us  Add to Digg  Add to Reddit 
所有评论 (0)
暂无评论
 
评注功能仅针对注册用户开放,请您登录
您对该条目有什么异议,请填写以下表单,管理员会尽快联系您。
内 容:
Email:  *
单位:
验证码:   刷新
您在IR的使用过程中有什么好的想法或者建议可以反馈给我们。
标 题:
 *
内 容:
Email:  *
验证码:   刷新

Items in IR are protected by copyright, with all rights reserved, unless otherwise indicated.

 

 

Valid XHTML 1.0!
Powered by CSpace