MICROCHEMICAL JOURNAL
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2020
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IF 3.594
Lulu Hao,Liying Zhao,Guangbao Li,Yingying Li,Ligang Ma,Yanju Liu,Wenbin Wang,Jinming Kong
Cytokeratin fragment antigen 21-1 (CYFRA 21-1) DNA is a significant biomarker with relation to non-small cell lung cancer. Here, we described an ultrasensitive electrochemical biosensor for detection of CYFRA 21-1 DNA (target DNA, tDNA) via surface-initiated reversible addition fragmentation chain transfer (SI-RAFT) polymerization-based DNA metallization signal amplification strategy. Specifically, 5′terminal modified thiol peptide nucleic acid (PNA) probes are immobilized on the gold electrode surface (by Au-S selfassembly) for the identification of specific tDNA sequences. After hybridization, the 4-cyano-4-(phenylcarbonothioylthio) pentanoic acid (CPAD) is tethered to the PNA/DNA duplexes by means of the recognized phosphate-Zr 4+ -carboxylate chemistry. Subsequently, in the situation of C 3 H 4 O as the monomer, SI-RAFT polymerization can bring numerous aldehyde groups sites for the subsequent silver mirror reaction. The acrolein-polymers act as reductant to reduce Ag + into Ag 0 , and then the silver particles are deposited on the polymer skeleton, which significantly amplifies the electrochemical signal. Under optimal conditions, the designed sensor for tDNA detection show a wide linear range from 100 aM to 1 nM, and the detection limit is as low as 0.89 aM. Moreover, the proposed biosensor exhibits prominent applicability for analysis of tDNA in serum samples and high selectivity for different mismatched oligonucleotide fragments. Given the proposed biosensor without extra natural enzyme or bio-labels, this method has enormous potential in bioanalysis.
研究领域:
生物技术
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医学诊断
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分子生物学
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产品关联
DOI
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
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2020
·Vol. 412
·期 11
·2413-2421
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IF 3.637
Wang Xia,Zhang Yawen,Zhao Liying,Wang Dazhong,Yang Huaixia,Kong Jinming
An ultrasensitive fluorescence biosensor for detecting cytokeratin fragment antigen 21-1 (CYFRA 21-1) DNA of non-small cell lung carcinoma (NSCLC) is designed using polysaccharide and activator regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP) signal amplification strategy. Thiolated peptide nucleic acid (PNA) is fixed on magnetic nanoparticles (MNPs) by a cross-linking agent and hybridized with CYFRA 21-1 DNA. Hyaluronic acid (HA) is linked to PNA/tDNA heteroduplexes in the form of carboxy-Zr 4+ -phosphate. Subsequently, multiple 2-bromo-2-methylpropionic acid (BMP) molecules are linked with HA to initiate ARGET ATRP reaction. Finally, a large number of fluorescein o-acrylate (FA) monomers are polymerized on the macro-initiators, and the fluorescence signal is significantly amplified. Under optimal conditions, this biosensor shows a significant linear correlation between the fluorescence intensity and logarithm of CYFRA 21-1 DNA concentration (0.1 fM to 0.1 nM), and the limit of detection is as low as 78 aM. Furthermore, the sensor has a good ability to detect CYFRA 21-1 DNA in serum samples and to recognize mismatched bases. It suggests that the strategy has broad application in early diagnosis by virtue of its high sensitivity and selectivity.
A novel and highly sensitive fluorescence biosensor for quantitatively detecting CYFRA 21-1 DNA via dual signal amplification of hyaluronic acid and ARGET ATRP reaction was developed. This proposed method has a low detection limit, wide detection range, high selectivity, and strong anti-interference.
研究领域:
生物技术
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癌症研究
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纳米技术
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分子生物学
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2019
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IF 9.518
Liying Zhao,Huaixia Yang,Xiaoke Zheng,Jinge Li,Lihe Jian,Weisheng Feng,Jinming Kong
Cytokeratin fragment antigen 21–1 (CYFRA 21–1) DNA is a crucial biomarker closely associated with non-small cell lung cancer. Here, we fabricated a novel electrochemical biosensor for ultrasensitive detection of CYFRA 21–1 DNA via polysaccharide and electrochemically mediated atom transfer radical polymerization (eATRP) dual signal amplification . Specifically, thiolated peptide nucleic acid (PNA) probes at 5′-terminals are immobilized on the gold electrode surface for specific recognition of CYFRA 21–1 DNA (tDNA). After hybridization, hyaluronic acid (HA) is linked to the hybridized PNA/DNA duplexes via the recognized carboxylate-Zr 4+ -phosphate chemistry . Then multiple initiators of the polymerization reaction are introduced via esterification reaction. Lastly, large numbers of electro-active monomers are successfully grafted from the initiation sites of functionalized HA by eATRP reaction, significantly amplifying the electrochemical signal. Under optimal conditions, the constructed sensor can detect as low as 9.04 aM tDNA. Further, this proposed biosensor can also be applied to the direct detection of double-stranded DNA (dsDNA), obtaining 0.12 fM as the detection limit. Besides, this strategy shows high selectivity for mismatched bases and excellent applicability for CYFRA 21–1 DNA detection in the serum samples. Given its high sensitivity, selectivity, ease of operation, low cost and environmental friendliness , this biosensor has considerable potential in early diagnosis and biomedical application .
研究领域:
生物化学
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癌症研究
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分析化学
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
TALANTA
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2020
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IF 5.339
Xiaofei Li,Yawen Zhang,Lulu Hao,Yanju Liu,Xia Wang,Huaixia Yang,Jinming Kong
Cytokeratin fragment antigen 21-1 (CYFRA 21-1) DNA is perceived as sensitive tumor marker for the diagnosis of non-small cell lung cancer and other tumor. Herein, linear chain poly(ε-caprolactone) (PCL) synthesized by ring-opening polymerization is applied to ultrasensitive label-free electrochemical impedance detection system for CYFRA 21-1 DNA. First, thiolated peptide nucleic acid (PNA) is self-assembled into the Au electrode surface through the formation of Au–S bonds, allowing the PNA to act as biomolecular probe and form PNA/DNA heteroduplex with the target DNA via specific hybridization. Then, PCL is conjugated to the immobilized DNA on the electrode via “carboxylate–Zr 4+ –phosphate” bridges. Finally, the electrochemical response of modified PNA/DNA/Zr 4+ /PCL electrode is determined by electrochemical impedance method to quantify of CYFRA 21-1 DNA. Under optimal conditions, this method exhibits highly sensitivity with a broad linear range (0.1 fM – 1 nM) ( R 2 = 0.995) and the limit of detection (LOD) is as low as 10.73 aM, which is equivalent to just 64 molecules in a 10 μL sample. What's more, the high selectivity , good anti-interference, label-free operation, and real-time monitoring in complex samples of the proposed strategy demonstrate its broad application for the early diagnosis and clinical monitoring.
研究领域:
生物医学工程
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分析化学
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分子生物学
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癌症诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICAL SCIENCES
·
2019
·Vol. 36
·期 6
·681-685
·
IF 1.618
Liu Wenwen,Ma Ligang,Guo Zhuangzhuang,Liu Tao,Liu Yanju,Wang Dazhong,Kong Jinming
We propose a novel, ultrasensitive and low-cost sensor using reversible addition-fragmentation chain transfer (RAFT) polymerization as a signal amplification strategy for the detection of CYFRA 21-1 DNA fragment, a tumor marker of non-small cell lung carcinoma. The peptide nucleic acid (PNA) probes were firstly immobilized on magnetic beads (MBs) to capture the CYFRA 21-1 DNA specifically. After hybridization, CPAD was tethered to the hetero duplexes through carboxylate-Zr 4+ -phosphate chemistry. Subsequently, a number of fluorescent tags were introduced to the heteroduplexes through RAFT polymerization, leading to an amplification of the fluorescence signal. The sensor demonstrates a low limit of detection (LOD) of 0.02 fM. It has great selectivity with respect to base mismatch DNA, and high anti-interference ability in normal human serum. Overall findings of the study suggest that proposed sensor holds enormous potential to be used as a tool for the early-stage diagnosis of lung cancers.
研究领域:
生物技术
·
癌症诊断
·
纳米技术
·
分析化学
✓ 已验证
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）