ICCAS OpenIR
Chemiluminescently labeled aptamers as the affinity probe for interaction analysis by capillary electrophoresis
Li, Hong-Yi; Deng, Qin-Pei; Zhang, De-Wen; Zhou, Ying-Lin; Zhang, Xin-Xiang
2010-07-01
Source PublicationELECTROPHORESIS
ISSN0173-0835
Volume31Issue:14Pages:2452-2460
AbstractAptamers 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.
KeywordAffinity Probe Aptamer Ce Chemiluminescence Interaction Analysis
DOI10.1002/elps.201000131
Indexed BySCI
Language英语
WOS IDWOS:000280709700021
PublisherWILEY-V C H VERLAG GMBH
Citation statistics
Cited Times:11[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.iccas.ac.cn/handle/121111/69192
Collection中国科学院化学研究所
Corresponding AuthorZhang, Xin-Xiang
AffiliationPeking Univ, BNLMS, Key Lab Bioorgan Chem & Mol Engn, Minist Educ,Inst Analyt Chem,Coll Chem, Beijing 100871, Peoples R China
Recommended Citation
GB/T 7714
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.
APA Li, Hong-Yi,Deng, Qin-Pei,Zhang, De-Wen,Zhou, Ying-Lin,&Zhang, Xin-Xiang.(2010).Chemiluminescently labeled aptamers as the affinity probe for interaction analysis by capillary electrophoresis.ELECTROPHORESIS,31(14),2452-2460.
MLA Li, Hong-Yi,et al."Chemiluminescently labeled aptamers as the affinity probe for interaction analysis by capillary electrophoresis".ELECTROPHORESIS 31.14(2010):2452-2460.
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