کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1246792 | 969768 | 2009 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
An aptameric molecular beacon-based “Signal-on” approach for rapid determination of rHuEPO-α
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کلمات کلیدی
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
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چکیده انگلیسی
The aim of this work is to describe the first example of aptameric molecular beacon (MB)-based probe for the detection of recombinant human erythropoietin (rHuEPO-α) in physiological buffer, using a novel 35 nt ssDNA aptamer (807-35 nt) originally isolated by Systematic Evolution of Ligands by Exponential enrichment (SELEX) technique in our laboratory. Both “Signal-on” and “Signal-off” MB modes were developed, respectively, in which the conformational alteration of aptamer before and after binding to rHuEPO-α can be demonstrated in terms of the correspondingly fluorescent changes. Comparing with “Signal-off” mode, “Signal-on” mode provided higher sensitivity, while with the addition of target rHuEPO-α, quenching between fluorescent 807-35 nt aptamer (F-Apt) and a short quencher-labeled complementary sequence (QDNA) was disturbed by the specific binding between rHuEPO-α and F-Apt. QDNA was thus loosened and released from F-Apt, leading to a consequently full fluorescent restoration. Systematic optimization of parameters in “Signal-on” mode were carried out, the choice of QDNA length, the hybridization site of a small supplementary DNA (SDNA) stabilizer, and the existence of Mg2+ cation played essential roles for the performance characterization. A convenient and sensitive determination of rHuEPO-α with a LOD of 0.4 nM was achieved.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Talanta - Volume 80, Issue 2, 15 December 2009, Pages 985-990
Journal: Talanta - Volume 80, Issue 2, 15 December 2009, Pages 985-990
نویسندگان
Zhaoyang Zhang, Lei Guo, Jijun Tang, Xingjie Guo, Jianwei Xie,