Amperometric sandwich-type aptasensor based on peroxidase mimetic Au@Pt for the carcinoembryonic antigen

For sensitive analysis of cancer biomarker carcinoembryonic antigen (CEA), an amperometric sandwich-type aptasensor is proposed based on a signal amplification strategy of Au@Pt bimetallic nanoprobes. As the excellent catalytic activity to hydrogen peroxide (H₂O₂), core-shell Au@Pt nanoparticles are employed as nanoprobes by conjugating directly with the secondary aptamer of CEA (Apt-II). Due to the synergic recognition effect of dual aptamers and the excellent catalytic activity of nanoprobes, this amperometric sandwich-type aptasensor for CEA exhibits high specificity and good sensitivity with a limit of detection of 0.31 ng/mL, along with a wide linear range from 0.1 ng/mL to 100 ng/mL.     

Facile and sustainable synthesis of sodium lignosulfonate derived carbon quantum dots for the detection of total Mn and ascorbic acid

A fluorescent nanoprobe based on sodium lignosulfonate derived carbon quantum dots (SLS-CQDs) was fabricated through a facile and sustainable one-step hydrothermal treatment. The obtained SLS-CQDs had excellent fluorescence properties and were applied to detect total Mn and ascorbic acid (AA). The fluorescence of SLS-CQDs could be effectively quenched by Mn based on a static quenching process and recovered by the addition of AA. An “on–off-on” fluorescent nanoprobe was established to determine total Mn and AA with the detection limits of 6.9 µg/L and 0.65 µM with the corresponding linear ranges of 0.25–2.25, 5–25 mg/L, and 5–110 µM, respectively. This nanoprobe could accurately detect total Mn in surface water and AA in Vitamin C effervescent tablet samples. The proposed method is simple and can be performed easily, indicating that SLS-CQDs might be used in environmental monitoring, food and drug testing.     

孔雀石绿的磁性免疫层析快速检测方法研究

开发了一种适用于现场快速检测孔雀石绿(MG)的免疫层析试纸条,在超顺磁性纳米微球上偶联MG单克隆抗体作为检测探针,分别将孔雀石绿完全抗原(MG-BSA)和羊抗鼠IgG喷涂于NC膜的T线和C线。结果发现,T线最佳喷涂量为0.25 mg/mL,抗体最佳偶联量为20μg,构建的试纸条可在25 min内实现养殖用水及鱼肉组织的定性与定量分析,检出限为1μg/mL,定量限分别为0.60,0.97 ng/mL,对无色孔雀石绿无交叉反应,批内、批间相对标准偏差均<12%;通过与HPLC和市售胶体金试剂盒对比发现,水样的回收率在82.0%~117.4%,灵敏度高于市售胶体金试剂盒。该孔雀石绿磁性免疫层析条适用于现场快速检测鱼肉或水体中的残留孔雀石绿。     

3,4-Dinitrobenzamide Functionalized CdTe/ZnTe Quantum Dots as a Nanoprobe for Imaging Glutathione S-Transferase in Living Cells

A novel fluorescent nanoprobe for glutathione S‐transferase (GST) has been developed by incorporating 3,4‐dinitrobenzamide (a specific substrate of GST) onto CdTe/ZnTe quantum dots. The probe itself displays a low background signal due to the strong quenching effect of the electron‐withdrawing unit of 3,4‐dinitrobenzamide on the quantum dots. However, GST can efficiently catalyze the nucleophilic substitution of reduced glutathione on the p‐nitro group of the nanoprobe, leading to a large fluorescence enhancement. Most notably, this enhancement shows high selectivity and sensitivity towards GST instead of the other biological substances. With this nanoprobe, a simple fluorescence imaging method for intracellular GST has been established, and its applicability has been successfully demonstrated for imaging GST in different living cells, which reveals that A549 cells express GST about 3 times higher than NIH‐3T3 and Hela cells.     

pH‐sensing nanostar probe using surface‐enhanced Raman scattering (SERS): theoretical and experimental studies

Local pH environment has been considered to be a potential biomarker for tumor diagnosis because solid tumors contain highly acidic environments. A pH‐sensing nanoprobe based on surface‐enhanced Raman scattering (SERS) using nanostars under near‐infrared excitation has been developed for potential biomedical applications. To theoretically investigate the effect of protonation state on SERS spectra of p‐mercaptobenzoic acid (pMBA), we used the density functional theory (DFT) with the B3LYP functional to calculate Raman vibrational spectra of pMBA‐Au/Ag complex in both protonated and deprotonated states. Vibrational spectral bands were assigned with DFT calculation and used to investigate SERS spectral changes observed from experiment when varying pH value between five and nine. The SERS peak position of pMBA at ~1580 cm⁻¹ was identified to be a novel pH‐sensing index, which has small but noticeable downshift with pH increase. This phenomenon is confirmed and well‐explained with theoretical simulation. The study demonstrates that SERS is a sensitive tool to monitor minor structural changes due to local pH environment, and DFT calculations can be used to investigate Raman spectra changes associated with minor differences in molecular structure. Copyright © 2013 John Wiley & Sons, Ltd.     

乙二醛诱导细胞凋亡的近红外表面增强拉曼光谱

将60 nm金纳米粒子导入到活的人骨肉瘤细胞中, 用近红外表面增强拉曼散射(SERS)技术获取细胞内化学成分的高分辨SERS信息. 对正常活性细胞和乙二醛诱导的凋亡细胞的比较研究表明, 对于正常活性的细胞, 金纳米探针主要分布在细胞质内(围绕细胞核), 而凋亡细胞内的金纳米探针的分布较为均匀, 在遍布凋亡细胞内的各个位置包括细胞表面均容易找到DNA片段的信息.     

Synthesis of PAMAM-g-PEG-g-DS-g-RB@IO towards dual-modal imaging in atherosclerosis

This study aimed to synthesize a dual-modal magnetic resonance imaging (MRI)/fluorescence imaging (FLI) nanoprobe in order to detect atherosclerosis. Herein, we had prepared the polyamidoamine-graft-poly (ethylene glycol) (PAMAM-g-PEG) as the carrier. Dextran sulfate (DS) and Rhodamine B (RB) were grafted to PAMAM-g-PEG continuously through reductive amination and amidation reaction to synthesize PAMAM-g-PEG-g-DS-g-RB. The structure of PAMAM-g-PEG-g-DS-g-RB was characterized through nuclear magnetic resonance (¹H-NMR) and Fourier transform infrared spectroscopy (FTIR). A new water-soluble superparamagnetic iron oxide nanoparticles (IONPs) has been synthesized through simple ligand exchange between the iron oxide nanaparticles and PAMAM-g-PEG-g-DS-g-RB. The analysis of transmission electron microscopy (TEM) indicated that micelles were well dispersed in water and had uniform sizes. The result of thermogravimetric analysis (TGA) proved that about 83% (mass fraction) polymers were coated on the surface of IONPs. The MRI in vitro evaluation demonstrated a high R2 value (130.8 mM^?1s^?1) to be served as a T2-weighted contrast agent. The cell counting kit (CCK) assay showed no significant toxicity in RAW264.7. The above results confirmed that PAMAM-g-PEG-g-DS-g-RB@IO could play an significant role of MRI and FLI in the atherosclerosis.     

Development of Nanosensors and Bioprobes

We describe the development and application of nanosensors having bioreceptor probes for bioanalysis. The nanoprobes were fabricated with optical fibers pulled down to tips having distal end sizes of approximately 30–60nm. The use of two different types of receptors was investigated. Fiberoptic nanoprobes were covalently bound either with bioreceptors, such as antibodies, or with other receptors, such as cyclodextrins that are selective for the size and chemical structure of the analyte molecules. Theoretical calculations were performed to model the binding of beta-cyclodextrin with pyrene and 5,6-benzoquinoline, and to illustrate the possibility of comparing experimental data with theoretical data. The antibody-based nanoprobe was used for in situ measurements of benzopyrene tetrol in single cells. The performance of the nanosensor is illustrated by intracellular measurements performed on a rat liver epithelial cell line (Clone 9) used as the model cell system. The usefulness and potential of these nanotechnology-based biosensors in biological research and applications are discussed.     

纳米荧光探针用于核酸分子的检测及成像研究

核酸,包括脱氧核糖核酸和核糖核酸,在生物的生长、发育、突变、炎症、癌症等正常或异常的生命活动中发挥着重要的作用,它们的异常表达与多种疾病的发生、发展也密切相关.因此,发展准确、有效的方法实现核酸分子的检测,对深入探究核酸的功能调控以及相关疾病的早期检测与治疗都具有重要的意义.荧光检测法与荧光成像技术具有灵敏度高、时空分辨率高等优点,为实时、准确的检测核酸分子提供了有力的工具.本文着重综述了近年来发展的纳米荧光探针用于疾病相关核酸分子的检测与细胞和活体成像工作的研究进展,最后提出了进一步构建新型纳米荧光探针用于核酸检测面临的挑战、未来发展方向与展望.