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Multiple Analytes Detection Based on Gold Nanoparticles and Fluorescent Oligonucleotides in One-Pot

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## Multiple Analytes Detection Based on Gold Nanoparticles and Fluorescent Oligonucleotides in One-Pot

###### 通讯作者: CHENG Sheng, chengsh@ustc.edu.cn ; YU Yongqiang, cjr.yuyongqiang@vip.163.com ; HU Jinming, jmhu@ustc.edu.cn
• 中图分类号: O652.7

## Multiple Analytes Detection Based on Gold Nanoparticles and Fluorescent Oligonucleotides in One-Pot

###### Corresponding author: CHENG Sheng, chengsh@ustc.edu.cn ;YU Yongqiang, cjr.yuyongqiang@vip.163.com ;HU Jinming, jmhu@ustc.edu.cn
• CLC number: O652.7

• 摘要: 利用具有不同荧光修饰的寡聚核苷酸(DNA)作为响应基团、纳米金粒子作为淬灭基团构建了多组分“turn-on”传感器，从而达到制备便捷、检测快速且灵敏，同时尽量减少干扰的目的。由于不同检测物识别序列的互补链上修饰的荧光基团不同，可通过不同的荧光达到对不同检测物检测的目的。利用此原理构建了针对腺苷和钾离子的传感器，对腺苷的检出限达387.9 nmol/L，检测范围0～15 μmol/L，通过调节识别序列检测范围可提高到mmol/L级别；对钾离子的检出限达1.6 μmol/L，检测范围2～6 μmol/L。
• 图 FIG. 173.  FIG. 173.

Figure FIG. 173..

Figure 1.  A schematic representation of multiple detection by fluorescent DNA and AuNPs

Figure 2.  TEM images of AuNPs at different magnifications

Figure 3.  (a) Fluorescent signals at 520 nm with (red) and without (black) 10 mmol/L adenosine in response to varying aptcpl concentrations; (b) Linear relationship between fluorescence emission intensity at 520 nm and adenosine concentration in the range of 0–15 μmol/L (Error bars represent the standard deviation of three measurements)

Figure 4.  Fluorescence spectra of detection system in response to adenosine with concentrations ranging from 1 mmol/L to 20 mmol/L

Figure 5.  Fluorescence emission at 520 nm as a function of adenosine concentrations (Error bars represent the standard deviation of three measurements)

Figure 6.  (a) Fluorescence spectra of detection system in response to potassium ions concentrations from 0 to 8.0 μmol/L; (b) Fluorescence emission at 605 nm as a function of potassium ion concentrations

Figure 7.  (a) Fluorescence spectra of detection system in response to potassium ions concentrations from 0.3 μmol/L to 6.0 μmol/L. (b) Linear relationship between fluorescence emission intensity at 605 nm and potassium ions concentrations in the range of 2−6 μmol/L (Error bars represent the standard deviation of three measurements)

Figure 8.  Specificity of multiple detection system over analogues: adenosine, uridine, cytidine and guanosine (all 3 mmol/L)

Figure 9.  Specificity of multiple detection system over analogues: potassium ions (3 μmol/L), lithium ions and sodium ions (1 mmol/L for both)

•  [1] 谷梦鑫 , 张良顺 , 林嘉平 . DNA非均匀功能化纳米粒子的可编程自组装行为. 功能高分子学报, 2020, 33(3): 269-274. doi: 10.14133/j.cnki.1008-9357.20190429003 [2] 李淼 , 楼一层 . 基于脂质体-DNA复合体的“条形码分子开关”及其生物识别. 功能高分子学报, 2014, 27(3): -. [3] 李小芳1 , 冯小强1 , 杨声2 . 羧甲基壳聚糖-Cu (Ⅱ)配合物的抑菌活性及其与DNA的相互作用. 功能高分子学报, 2014, 27(1): -.

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##### 出版历程
• 收稿日期:  2019-01-24
• 网络出版日期:  2019-09-09
• 刊出日期:  2020-02-01

## Multiple Analytes Detection Based on Gold Nanoparticles and Fluorescent Oligonucleotides in One-Pot

###### 通讯作者: HU Jinming, jmhu@ustc.edu.cn
• 1. School of Chemistry and Chemical Engineering, Hefei Normal University, Hefei 230061, China
• 2. Department of Radiology, The First Affiliated Hospital of Anhui Medical University, Hefei 230022, China
• 3. Instrumental Analysis Center, Hefei University of Technology, Hefei 230009, China
• 4. Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026, China

### English Abstract

• Sensor is a concept that is inspired by biological system which can respond to stimuli and generate corresponding outputs. It is extremely useful in the fields of research and practical applications, including diagnostics in biomedicine, monitoring in environment and quality control in industry[1, 2]. Up to now, a number of studies on single analyte detection have been reported, and significant outcomes have been obtained for the constructions of single analyte sensors[3-5]. However, the simultaneous detection of multiple analytes is still challenging, because multiple detection procedures should be performed under the same conditions at the same time while generate different signals. This is pretty important in biological system where the synergistic or antagonistic effects of different substrates will have enormous effects on human body. Therefore, simultaneous monitor or detection of different substances is of great importance.

Ghosh et al[6] reported the detection of dopamine and ions simultaneously using trypsin mediated gold nanoclusters as signal reporters. The detection limits for carbidopa, dopamine, Cu2+, Co2+ and Hg2+ were 6.5, 0.14, 5.2, 0.007 8 nmol/L and 0.005 nmol/L, respectively. However, this method was based on a fluorescence turning off mechanism, leading to negative effects on the results. A tripodal receptor containing both nitrogen and oxygen binding sites was investigated to bind Cu2+ selectively with a detection limit of 4.6 μmol/L. The receptor-Cu2+ complex presented good recognition capacity towards ${\rm Br}^{-}$ which can be used as a potential chemosensor for multiple analytes detection[7]. This sensor only detected ${\rm Br}^{-}$ in case the receptor-Cu2+ complex was formed, therefore, the simultaneous detection is limited by the adding sequence. Wang et al[8] synthesized a terbium metal-organic framework (Ln-MOF) sensor that can not only detect Fe3+, Cr2O72−, CrO42−, ${\rm MnO}_4^{\;\;-}$, but also monitor nitromethane as well as trace amounts of nitro-aromatic compounds in aqueous solutions. Cyclodextrin modified supramolecular functional polymers were synthesized using molecular imprinting technique which can simultaneously recognize nitrophenol and bisphenol. The system presented good selectivity in practical water detection. However, the synthetic process prevented the large-scale production of the polymers[9]. A phospholipid removal micro-elution solid phase extraction method for sample preparation was developed for liquid chromatography coupled with mass spectroscopy (LC-MS)/mass spectroscopy (MS) instrument which can detect six antipsychotics simultaneously[10]. Gold nanoparticles (AuNPs) and nanostructures were also investigated for multiple analytes detection[11]. Good effects were arrived, but the detection was relied on expensive instrumentations.

To construct a biocompatible sensor, the selection of sensing element is a critical step. Antibodies, due to their high specificity, have long been utilized for sensing. However, the production and stability have limited their large-scale preparations and applications[12]. Nucleic acids, also one kind of biological molecules, are found to specifically bind with targets and can be separated and sequenced using Systematic Evolution of Ligands by Exponential Enrichment (SELEX) technique[13]. Since nucleic acids have higher stability and can be synthesized in vitro, a new name aptamer for this kind of nucleic acids with high affinity for target binding has been established since it was first discovered in 1990s[14, 15].

Different strategies were adopted for the construction of multiple detection. Liu et al[16] advanced a new and general method utilizing the disassembly of AuNPs during encountering targets. Luo et al[17] immobilized DNA modified with fluorophores with different emission wavelengths onto polystyrene for multiple detections. To minimize strands for one target detection, Li et al[18] used two-strand linking AuNPs strategy for multiple analyses. However, the simultaneous modification of different strands onto the same nanoparticle led to the problem of uneven arrangement of DNA with different strand lengths. Liu et al[19] explored one single strand DNA with fluorescence label for simultaneous detection of two heavy ions. This method was much simpler but had little generality. Mancuso et al[20] utilized the aggregations of AuNPs and silver nanoparticles as multiple sensing signals which had no overlapping spectra to distinguish the signals. Utilizing the porous characteristics of metal-organic frameworks (MOFs), electro-active dyes were capped using dsDNA as the capper. The surfaces of MOFs were modified with aptamers that could recognize different tumor biomarkers which would trigger the release of signals from MOFs[21]. The resulting sensor showed a detection limit as low as 3.6 fmol/L. Desirable progress has been made, yet multiple strands modified onto different nanoparticles with different fluorescences on each strand has not been explored.

In our previous work, the single detection of biological elements has been achieved, such as the monitor of pH[22], adenosine[23], ATP[24-26], lead[27], potassium ions[28], ochratoxin A[29] and saxitoxin[30, 31]. To pave the way for future practical application, simultaneous detection of adenosine and potassium ions in one-pot was investigated in this study. Utilizing the special recognition capability of aptamer, potassium and adenosine aptamers are linked onto AuNPs. Another partly complementary strand with fluorescence label can base pair with aptamer in case the target cannot induce fluorescence quenching, while fluorescence turning on when target is presented.

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