基于AuNPs/PANI-NF复合膜的电化学细胞传感器对姜黄素诱导HeLa细胞凋亡的抑制作用检测

以聚苯胺纳米纤维(PANI-NF)-纳米金(AuNPs)复合膜构建传感界面,在AuNPs/PANI-NF界面上修饰转铁蛋白(Tf),利用转铁蛋白与人宫颈癌细胞(HeLa)表面大量表达的转铁蛋白受体(TfR)之间的特异性识别作用,将细胞捕获到传感器界面,导致电化学阻抗值变化。利用电化学阻抗谱研究姜黄素对HeLa细胞的抑制作用。结果表明,随着药物浓度的增大和作用时间的延长,电化学阻抗值下降,表明姜黄素对细胞的抑制作用增强。电化学阻抗值的变化率与药物浓度和作用时间具有量效关系;电化学方法检测显示,姜黄素对HeLa细胞的抑制作用趋势与MTT法及倒置显微镜检测到的姜黄素对HeLa细胞的抑制作用结果相吻合,从而建立了一种检测药物对细胞抑制作用的新方法。该方法具有简便、灵敏度高、费用低廉等优点。     

点击功能化的叶酸受体靶向荧光纳米探针用于肿瘤细胞成像

采用点击化学偶联法对荧光二氧化硅纳米粒子表面进行叶酸功能化修饰,构建了一种叶酸受体靶向的荧光纳米探针,并成功用于肿瘤细胞的成像研究.首先通过St?ber法制备包裹钌联吡啶的荧光二氧化硅纳米粒子(RSiNPs),然后利用叠氮化硅烷偶联剂(Az-PTES)的水解反应在其表面引入叠氮基团,最后通过点击化学反应将炔丙基叶酸衍生物偶联到粒子表面.利用红外光谱对其偶联前后的叠氮基特征峰(2105 cm-1)进行表征,证实了叶酸功能化的荧光纳米探针(RSiNPs-Folate)已被成功制备.在生理pH条件下,以458 nm为激发波长,RSiNPs-Folate在601 nm处发射较强的红色荧光,且光稳定性较好.细胞成像结果表明,这种叶酸受体靶向的荧光纳米探针能够有效地标记叶酸受体呈阳性的人宫颈癌细胞(HeLa),而叶酸受体呈阴性的人肺癌细胞(A549)未观察到明显的荧光.叶酸竞争性结合实验证明了这种叶酸受体介导的肿瘤细胞成像机制.此探针能够实现混合细胞体系中HeLa细胞的选择性识别与荧光成像.与酰胺化反应偶联叶酸相比,这种点击功能化的纳米探针的合成方法简单、反应条件温和、产率高,可用于不同肿瘤细胞的荧光标记与成像.     

基于生物条形码的粘蛋白和特定细胞电化学发光适配体生物传感方法研究

本文基于适配体识别和生物条形码放大策略,以MCF-7细胞和粘蛋白(MUC1)为目标物,MUC1的特异性适配体(rcDNA)为分子识别物质,Ru(phen)₃²⁺为信号物质,rcDNA通过巯基自组装于金电极表面作为传感界面,发卡DNA(hpDNA)和rcDNA通过巯基自组装在金纳米粒子(AuNP)表面合成的hpDNA/AuNP/rcDNA为生物条形码探针,建立了测定MUC1和特定细胞的电化学发光适配体生物传感新方法.当目标物被传感界面上的rcDNA俘获后,进而与生物条形码探针形成夹心复合物,Ru(phen)32+嵌入hpDNA中.在共反应剂的存在和+0.95 V恒电位下测量电化学发光强度.电化学发光强度的变化值与MUC1和MCF-7细胞浓度的对数在4~800 pg/mL和30~5.0×10^(4 )cells/mL范围内呈良好的线性关系,检出限分别为0.5 pg/mL和9 cells/mL.将该法应用于监测两种物质刺激下MCF-7细胞表面MUC1含量的变化,发现芹黄素刺激下细胞表面MUC1含量降低,而过氧化氢刺激下细胞表面MUC1含量不变.     

基于功能纳米材料的灵敏生物传感策略

21世纪的第一个十年被称为＂传感的十载＂.功能纳米材料为灵敏的生物传感器件（包括光学和电生物传感）的制备提供了优秀的平台.这方面的大多数工作主要聚焦于不同纳米材料的生物功能化,例如金属纳米粒子、半导体纳米粒子和碳纳米粒子,功能化方式包括物理吸附、静电结合、特异性识别或共价键合.这些生物功能化纳米材料可以用作催化剂、电导体、光发射剂、载体或示踪剂,以获取被放大的检测信号、稳定的识别探针或生物传感界面.设计的信号放大策略已经极大地促进了不同领域中稳定、特异、具有选择性和灵敏的生物传感器的发展.本文介绍了基于功能纳米材料的一些生物传感新原理和检测新策略,也讨论了纳米材料的生物功能化方法和生物传感在蛋白质的免疫分析、DNA检测、糖分析和细胞传感中的应用.     

荧光无机/有机杂化纳米分子口袋的构建

利用能量匹配原则将无机半导体ZnO纳米粒子与有机半导体四苯基卟啉四羧酸(TCPP)分子进行组装, 在无机-有机组分之间的界面构建了杂化纳米分子口袋ZNPs-TCPP, 该纳米分子口袋对四苯基卟啉(TPP)分子具有高选择性识别功能, 同时其荧光增强了5倍以上, 并成功地将该杂化纳米分子口袋用于肺腺癌细胞(A549)的细胞成像.     

稀土掺杂LaF3超小发光纳米晶的合成及癌细胞靶向上转换发光成像研究

采用溶剂热法合成了超小尺寸三氟化镧（LaF₃）纳米晶. 利用XRD、TEM等对其进行结构与形貌表征,结果显示制得的纳米颗粒为LaF₃纳米晶,且具有高度均一的尺寸分布和良好的结晶度,颗粒尺寸为6±0.4 nm. 通过共掺杂镱铒两种稀土元素,所得材料在近红外光激发下（980 nm）可发射出明亮的绿光;发光光谱显示,在524,544,655 nm等位置有较强的发射峰. 利用聚琥珀酰亚胺高分子（PSI）对油溶性的纳米颗粒表面进行功能化修饰,将其成功转移水相. 利用叶酸对其进行进一步生物标记,可与肝癌细胞表面叶酸受体特异性识别,成功地实现了肝癌细胞上转换绿色发光成像研究.     

基于DNA纳米结构的传感界面调控及生物检测应用

生物传感技术在环境、安全和医学诊断等应用中具有重要意义。如何精确调控自组装界面上生物识别探针与界面的相互作用来提高生物传感的性能则是其中的关键问题。常规界面组装过程中,DNA等生物分子往往在界面形成非均一的自组装层,分子结合能量壁垒高,识别效率低。我们通过构建有序DNA纳米结构,发展了纳米尺度精确调控界面性质的方法。通过在界面上形成以熵驱动主导的均匀自组装层,增加探针分子间的有效距离,并通过精确调控界面上DNA纳米结构的尺寸,显著提高界面DNA杂交效率与速率。我们在DNA四面体上修饰不同的生物识别分子(DNA、抗体、核酸适配体等),可构建通用检测平台,实现对核酸、蛋白、小分子及细胞的高灵敏检测,并且在复杂样本中同样保持了优异的检测性能。在此基础上,我们将四面体三维结构探针应用于细胞内以及活体检测,研究了DNA四面体在细胞内的运输途径及靶向定位方式,并实现对细胞内ATP分布的传感成像及小鼠体内肿瘤组织的靶向成像,有望发展活体生物传感的新探针。     

靶向性纳米金探针对宫颈癌细胞的激光扫描共聚焦散射成像

制备了粒径均一的纳米金颗粒, 再对其表面进行叶酸修饰, 制得具有靶向性的纳米金探针. 利用激光扫描共聚焦显微镜(LSCM), 对靶向性纳米金的细胞特异性散射成像进行研究. 实验结果表明, 人宫颈癌细胞(Hela)对纳米金-叶酸的摄取作用强于对纳米金的摄取, 但随着时间的延长, 两者的差别逐渐减小. 表明在适当的时间内纳米金-叶酸探针对宫颈癌细胞具有良好的靶向性.     

界面聚合法合成手性掺杂剂诱导螺旋形聚苯胺纳米纤维

以手性试剂D-樟脑磺酸(D-CSA)和L-樟脑磺酸(L-CSA)为掺杂剂和构象诱导剂,采用界面聚合法合成了螺旋形聚苯胺纳米纤维。通过FESEM、TEM、FTIR和UV-Vis吸收光谱等测试技术对螺旋形聚苯胺纳米纤维结构进行了表征。结果表明,所得聚苯胺纤维具有螺旋形构象,形貌均一,平均直径约为50nm,长度为300～600nm,具有较高的长径比(6:1～12:1)。在水溶液中,聚苯胺纳米纤维以伸展的螺旋形分子链构象存在,调节溶液的pH值,螺旋形聚苯胺纳米纤维表现出可逆的掺杂和脱掺杂性质。循环伏安(CV)测试表明,螺旋形聚苯胺纳米纤维在0.5mol/LHCl溶液中表现出良好的电化学活性。     

基于纳米探针的肿瘤细胞电化学传感研究进展

纳米材料在纳米尺度下具有独特的光、电、磁等性质,已广泛应用于细胞电化学传感的研究,该研究工作提高了肿瘤细胞检测的灵敏度与特异性,为癌症的早期诊断与治疗提供了新的方法与思路。本文综述了基于纳米探针进行肿瘤细胞电化学传感研究的工作,总结了纳米电化学细胞传感技术在肿瘤细胞固定、识别、检测及诱导凋亡等方面的研究进展,并对未来研究动向进行了展望。     

Electrochemical current rectifier as a highly sensitive and selective cytosensor for cancer cell detection

Signal amplification originating from electrochemical current rectifier (ECR) was firstly applied to construct a cytosensor for rapid and non-invasive detection of folate receptor-rich cancer cells with high sensitivity. It exhibits a broad linear range with a detection limit as low as 10 cells mL(-1) even in the presence of a large number of normal cells.     

Label-free electrochemical aptasensor constructed by layer-by-layer technology for sensitive and selective detection of cancer cells

Here, a cytosensor was constructed with ferrocene-appended poly(allylamine hydrochloride) (Fc-PAH) functionalized graphene (Fc-PAH-G), poly(sodium-p-styrenesulfonate) (PSS) and aptamer (AS1411) by layer-by-layer assembly technology. The hybrid nanocomposite Fc-PAH-G not only brings probes on the electrode and also promotes electron transfer between the probes and the substrate electrode. Meanwhile, LBL technology provides more effective probes to enhance amplified signal for improving the sensitivity of the detection. While AS1411 forming G-quardruplex structure and binding cancer cells, the current response of the sensing electrode decreased due to the insulating properties of cellular membrane. Differential pulse voltammetry (DPV) was performed to investigate the electrochemical detection of HeLa cells attributing to its sensitivity of the current signal change. The as-prepared aptasensor showed a high sensitivity and good stability, a widely detection range from 10 to 10⁶ cells/mL with a detection limit as low as 10 cells/mL for the detection of cancer cells.     

基于AuNPs/PDDA-GO纳米复合物的电化学免疫传感器的构建及对SirT1蛋白的检测

基于AuNPs/PDDA-GO纳米复合物制备了一种新型电化学免疫传感器, 并将其用于SirT1的检测. 首先, 在电极表面修饰复合材料AuNPs/PDDA-GO, 然后将目标蛋白SirT1固定到修饰了AuNPs/PDDA-GO的电极表面, 再通过特异性免疫反应结合一抗(Ab₁)和辣根过氧化酶标记的二抗分子(HRP-Ab₂), 最后用示差脉冲伏安法检测电流信号, 实现了对SirT1蛋白水平的测定. 在优化的实验条件下, SirT1蛋白的浓度在0.1~100 ng/mL范围内与响应电流呈良好线性关系, 检出限为0.029 ng/mL.     

An electrochemical method to detect folate receptor positive tumor cells

This paper reports an electrochemical method to detect folate receptor positive tumor cells by making use of the interaction between folic acid immobilized on gold nanoparticles and its receptor over-expressed on tumor cell membrane. Experimental results have shown that a gold electrode modified with folic acid functionalized gold nanoparticles can clearly denote folate receptor positive tumor cells, such as ovarian tumor cells and human cervical cancer cells. So, electrochemical technique has been introduced for cancer cells detection and a simple method to detect folate receptor positive tumor cells has been developed.     

Dual Amplified Electrochemical Immunosensor for Hepatitis B Virus Surface Antigen Detection Using Hemin/G‐Quadruplex Immobilized onto Fe3O4‐AuNPs or (Hemin‐Amino‐rGO‐Au) Nanohybrid

A sensitive electrochemical immunosensor for Hepatitis B virus surface antigen (HBsAg) detection was fabricated based on hemin/G‐quadruplex interlaced onto Fe₃O₄‐AuNPs or hemin ‐amino‐reduced graphene oxide nanocomposite (H‐amino‐rGO‐Au). G‐quadruplex DNAzyme, which is composed of hemin and guanine‐rich nucleic acid, is an effective signal amplified tool for its outstanding peroxidase activity and Fe₃O₄‐AuNPs or (H‐amino‐rGO‐Au) nanocomposites with quasi‐enzyme activity provide appropriate support for the immobilization of hemin/G‐quadruplex. The target protein was sandwiched between the primary antibody immobilized on the GO and secondary antibody immobilized on the Fe₃O₄‐AuNPs or (H‐amino‐rGO‐Au) nanocomposites and glutaraldehyde was used as linking agent for the immobilization of primary antibody on the surface of GO. Both Fe₃O₄‐AuNPs and H‐amino‐rGO‐Au nanocomposite and also hemin/G‐quadruplex can cooperate the electrocatalytic reduction of H₂O₂ in the presence of methylene blue as mediator. The proposed immunosensor has a wide linear dynamic range of 0.1 pg/ml to 300 pg/ml with a detection limit of 60 fg/ml when Fe₃O₄‐AuNPs was used for immobilization of hemin/G‐quadruplex, while the dynamic range and DL were 0. 1–1000 pg/mL and 10 fg/mL, respectively in the presence of H‐amino‐rGO‐ Au nanocomposite as platform for immobilizing of hemin/G‐quadruplex. The proposed immunosensor was also used for analysis of HBsAg in spiked human serum samples with satisfactory results.     

A repeatable assembling and disassembling electrochemical aptamer cytosensor for ultrasensitive and highly selective detection of human liver cancer cells

In this work, a repeatable assembling and disassembling electrochemical aptamer cytosensor was proposed for the sensitive detection of human liver hepatocellular carcinoma cells (HepG2) based on a dual recognition and signal amplification strategy. A high-affinity thiolated TLS11a aptamer, covalently attached to a gold electrode through Au–thiol interactions, was adopted to recognize and capture the target HepG2 cells. Meanwhile, the G-quadruplex/hemin/aptamer and horseradish peroxidase (HRP) modified gold nanoparticles (G-quadruplex/hemin/aptamer–AuNPs–HRP) nanoprobe was designed. It could be used for electrochemical cytosensing with specific recognition and enzymatic signal amplification of HRP and G-quadruplex/hemin HRP-mimicking DNAzyme. With the nanoprobes as recognizing probes, the HepG2 cancer cells were captured to fabricate an aptamer-cell-nanoprobes sandwich-like superstructure on a gold electrode surface. The proposed electrochemical cytosensor delivered a wide detection range from 1 × 10² to 1 × 10⁷ cells mL⁻¹ and high sensitivity with a low detection limit of 30 cells mL⁻¹. Furthermore, after the electrochemical detection, the activation potential of −0.9 to −1.7 V was performed to break Au–thiol bond and regenerate a bare gold electrode surface, while maintaining the good characteristic of being used repeatedly. The changes of gold electrode behavior after assembling and desorption processes were investigated by electrochemical impedance spectroscopy and cyclic voltammetry techniques. These results indicate that the cytosensor has great potential in disease diagnostic of cancers and opens new insight into the reusable gold electrode with repeatable assembling and disassembling in the electrochemical sensing.     

基于AuNPs/PANI/TNTs纳米复合材料的电化学检测妥布霉素的适配体传感器

本文提出了一种新型的检测妥布霉素的电化学适配体传感器,以差分脉冲伏安法（DPV）作为检测技术,亚甲基蓝作为电化学响应信号. 构建了以纳米复合材料金纳米粒子/聚苯胺/二氧化钛纳米管(AuNPs/PANI/TNTs)修饰玻碳电极的电极支架. 通过透射电子显微镜和X-射线光电子能谱对纳米复合材料进行详细的表征. 循环伏安图和电化学阻抗谱显示AuNPs/PANI/TNTs可以很好地增加电极的界面传导性能. DPV结果显示电流密度的响应和妥布霉素浓度之间存在一个很好的线性关系,并且得到一个宽广的检测范围为0.5 μmol·L^-1到70 μmol·L^-1. 本文提出的适配体基的传感器有很好的重复性和稳定性,作为一个潜在的手段可以应用在生物分析和医疗诊断中.     

Dopamine-loaded Liposomes-amplified Electrochemical Immunoassay Based on MXene (Ti3C2)−AuNPs

A simple and novel electrochemical immunoassay based on MXene (Ti₃C₂)−Au nanoparticles (AuNPs) was designed for sensitive screening of a disease-related biomarker, prostate-specific antigen (PSA), by using dopamine-loaded liposomes (DLL) for signal amplification. The system involves two parts, namely, sandwich-type immunoreaction to capture DLL and electrochemical measurement of dopamine. The target PSA can cause a specific antigen-antibody reaction and DLL are enriched in the enzyme-labeled pores. After Triton X-100 is injected into the detection cell, the carried DLL was quickly cracked to release dopamine wrapped in the cavity. A nanocomposite consisting of MXene (Ti₃C₂) support to immobilize Au nanoparticles (Ti₃C₂−Au) was utilized to modify a glassy carbon electrode, which gives a strongly enhanced differential pulse voltammetric (DPV) signals for dopamine. In this case, the change of DPV signal depends on the amount of dopamine released by liposomes, which is further positively correlated with the concentration of the analyte PSA. Combining the of MXene (Ti₃C₂)−AuNPs nanomaterials (large specific surface area, excellent electrical conductivity, and good electrocatalytic properties) with the liposome signal amplification strategy, the electrochemical immunoassay exhibited excellent performance toward PSA determination with a broad linear range of 1 pg/mL to 50 ng/mL and limit of detection down to 0.31 pg/mL (S/N=3) under the optimized testing conditions. High specificity for PSA over other disease-related biomarkers and acceptable nanocomposite/electrode stability were acquired. The excellent analytical performance shows that the current strategy provides an effective detection platform for clinical sample analysis.