Highly amplified detection of visceral adipose tissue-derived serpin (vaspin) using a cognate aptamer duo

被引:51
作者
Raston, Nurul Hanun Ahmad [1 ]
Gu, Man Bock [1 ]
机构
[1] Korea Univ, Coll Life Sci & Biotechnol, Seoul 136713, South Korea
基金
新加坡国家研究基金会;
关键词
SERINE-PROTEASE INHIBITOR; SENSITIVE DETECTION; OBESITY; ADIPOCYTOKINE; THROMBIN; ANTIBODY; SITES; BIND;
D O I
10.1016/j.bios.2015.03.042
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
A cognate aptamer duo for visceral adipose tissue-derived serpin (vaspin) which distinctively bind to two different sites on vaspin with high affinity and specificity were successfully developed by using graphene oxide-based systematic evolution of ligands by exponential enrichment (GO-SELEX), which offers immobilization-free screening of aptamers. The specific and simultaneous bindings of this aptamer duo (V1 and V49 aptamers) to the different sites of vaspin were confirmed by circular dichroism (CD) analysis and both sandwich-type surface plasmon resonance (SPR) and quantum dot labelled fluorescence imaging analysis (V1 aptamer serves as primary capturing aptamer and V49 aptamer as secondary signalling aptamer or vice versa). With this vaspin cognate aptamer duo on SPR platform, the detection of the target vaspin were improved to the limit of detection down to 3.5 ng/ml in buffer and 4.7 ng/ml in human serum samples. This cognate aptamer duo based biosensor could be utilized in the early diagnosis of type-2 diabetes. (C) 2015 The Authors. Published by Elsevier B.V.
引用
收藏
页码:261 / 267
页数:7
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