Co-reporter:Shi-Bo Cheng, Min Xie, Yan Chen, Jun Xiong, Ya Liu, Zhen Chen, Shan Guo, Ying Shu, Ming Wang, Bi-Feng Yuan, Wei-Guo Dong, and Wei-Hua Huang
Analytical Chemistry August 1, 2017 Volume 89(Issue 15) pp:7924-7924
Publication Date(Web):June 29, 2017
DOI:10.1021/acs.analchem.7b00905
Tumor metastasis is attributed to circulating tumor cells (CTC) or CTC clusters. Many strategies have hitherto been designed to isolate CTCs, but there are few methods that can capture and gently release CTC clusters as efficient as single CTCs. Herein, we developed a three-dimensional (3D) scaffold chip with thermosensitive coating for high-efficiency capture and release of individual and cluster CTCs. The 3D scaffold chip successfully combines the specific recognition and physically obstructed effect of 3D scaffold structure to significantly improve cell clusters capture efficiency. Thermosensitive gelatin hydrogel uniformly coated on the scaffold dissolves at 37 °C quickly, and the captured cells are gently released from chip with high viability. Notably, this platform was applied to isolate CTCs from cancer patients’ blood samples. This allows global DNA and RNA methylation analysis of collected single CTC and CTC clusters, indicating the great potential of this platform in cancer diagnosis and downstream analysis at the molecular level.
Co-reporter:Zi-He Jin, Yan-Ling Liu, Jing-Jing Chen, Si-Liang Cai, Jia-Quan Xu, and Wei-Hua Huang
Analytical Chemistry 2017 Volume 89(Issue 3) pp:
Publication Date(Web):December 28, 2016
DOI:10.1021/acs.analchem.6b04616
Carbon nanotube (CNT)-based flexible sensors have been intensively developed for physical sensing. However, great challenges remain in fabricating stretchable CNT films with high electrochemical performance for real-time chemical sensing, due to large sheet resistance of CNT film and further resistance increase caused by separation between each CNT during stretching. Herein, we develop a facile and versatile strategy to construct single-walled carbon nanotubes (SWNTs)-based stretchable and transparent electrochemical sensors, by coating and binding each SWNT with conductive polymer. As a polymer with high conductivity, good electrochemical activity, and biocompatibility, poly(3,4-ethylenedioxythiophene) (PEDOT) acting as a superior conductive coating and binder reduces contact resistance and greatly improves the electrochemical performance of SWNTs film. Furthermore, PEDOT protects the SWNTs junctions from separation during stretching, which endows the sensor with highly mechanical compliance and excellent electrochemical performance during big deformation. These unique features allow real-time monitoring of biochemical signals from mechanically stretched cells. This work represents an important step toward construction of a high performance CNTs-based stretchable electrochemical sensor, therefore broadening the way for stretchable sensors in a diversity of biomedical applications.
Co-reporter:Xin-Wei Zhang;Quan-Fa Qiu;Hong Jiang;Fu-Li Zhang;Dr. Yan-Lin Liu; Dr. Christian Amatore; Dr. Wei-Hua Huang
Angewandte Chemie International Edition 2017 Volume 56(Issue 42) pp:12997-13000
Publication Date(Web):2017/10/09
DOI:10.1002/anie.201707187
AbstractNanoelectrodes allow precise and quantitative measurements of important biological processes at the single living-cell level in real time. Cylindrical nanowire electrodes (NWEs) required for intracellular measurements create a great challenge for achieving excellent electrochemical and mechanical performances. Herein, we present a facile and robust solution to this problem based on a unique SiC-core–shell design to produce cylindrical NWEs with superior mechanical toughness provided by the SiC nano-core and an excellent electrochemical performance provided by the ultrathin carbon shell that can be used as such or platinized. The use of such NWEs for biological applications is illustrated by the first quantitative measurements of ROS/RNS in individual phagolysosomes of living macrophages. As the shell material can be varied to meet any specific detection purpose, this work opens up new opportunities to monitor quantitatively biological functions occurring inside cells and their organelles.
Co-reporter:Xin-Wei Zhang;Quan-Fa Qiu;Hong Jiang;Fu-Li Zhang;Dr. Yan-Lin Liu; Dr. Christian Amatore; Dr. Wei-Hua Huang
Angewandte Chemie 2017 Volume 129(Issue 42) pp:13177-13180
Publication Date(Web):2017/10/09
DOI:10.1002/ange.201707187
AbstractNanoelectrodes allow precise and quantitative measurements of important biological processes at the single living-cell level in real time. Cylindrical nanowire electrodes (NWEs) required for intracellular measurements create a great challenge for achieving excellent electrochemical and mechanical performances. Herein, we present a facile and robust solution to this problem based on a unique SiC-core–shell design to produce cylindrical NWEs with superior mechanical toughness provided by the SiC nano-core and an excellent electrochemical performance provided by the ultrathin carbon shell that can be used as such or platinized. The use of such NWEs for biological applications is illustrated by the first quantitative measurements of ROS/RNS in individual phagolysosomes of living macrophages. As the shell material can be varied to meet any specific detection purpose, this work opens up new opportunities to monitor quantitatively biological functions occurring inside cells and their organelles.
Co-reporter:Shan Guo, Jiaquan Xu, Min Xie, Wei Huang, Erfeng Yuan, Ya Liu, Liping Fan, Shibo Cheng, Songmei Liu, Fubing Wang, Bifeng Yuan, Weiguo Dong, Xiaolian Zhang, Weihua Huang, and Xiang Zhou
ACS Applied Materials & Interfaces 2016 Volume 8(Issue 25) pp:15917-15925
Publication Date(Web):June 6, 2016
DOI:10.1021/acsami.6b04002
Circulating tumor cells (CTCs) play a significant role in cancer diagnosis and personalized therapy, and it is still a significant challenge to efficiently capture and gently release CTCs from clinical samples for downstream manipulation and molecular analysis. Many CTC devices incorporating various nanostructures have been developed for CTC isolation with sufficient capture efficiency, however, fabricating such nanostructured substrates often requires elaborate design and complicated procedures. Here we fabricate a degradable zinc-phosphate-based hierarchical nanosubstrate (HZnPNS), and we demonstrate its excellent CTC-capture performance along with effective cell-release capability for downstream molecular analysis. This transparent hierarchical architecture prepared by a low-temperature hydrothermal method, enables substantially enhanced capture efficiency and convenient imaging. Biocompatible sodium citrate could rapidly dissolve the architecture at room temperature, allowing that 88 ± 4% of captured cells are gently released with a high viability of 92 ± 1%. Furthermore, antiepithelial cell adhesion molecule antibody functionalized HZnPNS (anti-EpCAM/HZnPNS) was successfully applied to isolate CTCs from whole blood samples of cancer patients, as well as release CTCs for global DNA methylation analysis, indicating it will serve as a simple and reliable alternative platform for CTC detection.
Co-reporter:Changliang Zhang, Jia-Quan Xu, Yu-Tao Li, Le Huang, Dai-Wen Pang, Yong Ning, Wei-Hua Huang, Zhiyong Zhang, and Guo-Jun Zhang
Analytical Chemistry 2016 Volume 88(Issue 7) pp:4048
Publication Date(Web):March 8, 2016
DOI:10.1021/acs.analchem.6b00374
The field-effect transistor (FET) biosensor has attracted extensive attentions, due to its unique features in detecting various biomolecules with high sensitivity and selectivity. However, currently used FET biosensors obtaining from expensive and elaborate micro/nanofabrication are always disposable because the analyte cannot be efficiently removed after detection. In this work, we established a photocatalysis-induced renewable graphene-FET (G-FET) biosensor for protein detection, by layer-to-layer assembling reduced graphene oxide (RGO) and RGO-encapsulated TiO2 composites to form a sandwiching RGO@TiO2 structure on a prefabricated FET sensor surface. After immobilization of anti-D-Dimer on the graphene surface, sensitive detection of D-Dimer was achieved with the detection limits of 10 pg/mL in PBS and 100 pg/mL in serum, respectively. Notably, renewal of the FET biosensor for recycling measurements was significantly realized by photocatalytically cleaning the substances on the graphene surface. This work demonstrates for the first time the development and application of photocatalytically renewable G-FET biosensor, paving a new way for G-FET sensor toward a plethora of diverse applications.
Co-reporter:Shi-Bo Cheng, Min Xie, Jia-Quan Xu, Jing Wang, Song-Wei Lv, Shan Guo, Ying Shu, Ming Wang, Wei-Guo Dong, and Wei-Hua Huang
Analytical Chemistry 2016 Volume 88(Issue 13) pp:6773
Publication Date(Web):June 11, 2016
DOI:10.1021/acs.analchem.6b01130
Effective isolation of circulating tumor cells (CTCs) has great significance for cancer research but is highly challenged. Here, we developed a microchip embedded with a three-dimensional (3D) PDMS scaffold by a quadratic-sacrificing template method for high-efficiency capture of CTCs. The microchip was gifted with a 3D interconnected macroporous structure, strong toughness, and excellent flexibility and transparency, enabling fast isolation and convenient observation of CTCs. Especially, 3D scaffold chip perfectly integrates the two main strategies currently used for enhancement of cell capture efficiency. Spatially distributed 3D scaffold compels cells undergoing chaotic or vortex migration in the channel, and the spatially distributed nanorough skeleton offers ample binding sites, which synergistically and significantly improve CTCs capture efficiency. Our results showed that 1–118 CTCs/mL were identified from 14 cancer patients’ blood and 5 out of these cancer patients showed 1–14 CTC clusters/mL. This work demonstrates for the first time the development of microchip with transparent interconnected 3D scaffold for isolation of CTCs and CTC clusters, which may promote in-depth analysis of CTCs.
Co-reporter:Hui Xie, Yu-Tao Li, Yong-Min Lei, Yan-Ling Liu, Meng-Meng Xiao, Chuan Gao, Dai-Wen Pang, Wei-Hua Huang, Zhi-Yong Zhang, and Guo-Jun Zhang
Analytical Chemistry 2016 Volume 88(Issue 22) pp:11115
Publication Date(Web):October 25, 2016
DOI:10.1021/acs.analchem.6b03208
An ultrasensitive and highly efficient assay for real-time monitoring of nitric oxide (NO) at single-cell level based on a reduced graphene oxide (RGO) and iron–porphyrin-functionalized graphene (FGPCs) field-effect transistor (FET) biosensor is reported. A layer-to-layer assembly of RGO and FGPCs on a prefabricated FET sensor surface through π–π stacking interaction allowed superior electrical conductivity caused by RGO, and highly catalytic specificity induced by metalloporphyrin, ensuring the ultrasensitive and highly specific detection of NO. The results demonstrated that the RGO/FGPCs FET biosensor was capable of real-time monitoring of NO in the range from 1 pM to 100 nM with the limit of detection as low as 1 pM in phosphate-buffered saline (PBS) and 10 pM in the cell medium, respectively. Moreover, the developed biosensor could be used for real-time monitoring of NO released from human umbilical vein endothelial cells (HUVECs) at single-cell level. Along with its miniaturized sizes, ultrasensitive characteristics, and fast response, the FET biosensor is promising as a new platform for potential biological and diagnostic applications.
Co-reporter:Wei Huang, Chu-Bo Qi, Song-Wei Lv, Min Xie, Yu-Qi Feng, Wei-Hua Huang, and Bi-Feng Yuan
Analytical Chemistry 2016 Volume 88(Issue 2) pp:1378
Publication Date(Web):December 27, 2015
DOI:10.1021/acs.analchem.5b03962
DNA methylation (5-methylcytosine, 5-mC) is the best characterized epigenetic mark that has regulatory roles in diverse biological processes. Recent investigation of RNA modifications also raises the possible functions of RNA adenine and cytosine methylations on gene regulation in the form of “RNA epigenetics.” Previous studies demonstrated global DNA hypomethylation in tumor tissues compared to healthy controls. However, DNA and RNA methylation in circulating tumor cells (CTCs) that are derived from tumors are still a mystery due to the lack of proper analytical methods. In this respect, here we established an effective CTCs capture system conjugated with a combined strategy of sample preparation for the captured CTCs lysis, nucleic acids digestion, and nucleosides extraction in one tube. The resulting nucleosides were then further analyzed by liquid chromatography–electrospray ionization–tandem mass spectrometry (LC-ESI-MS/MS). With the developed method, we are able to detect DNA and RNA methylation (5-methyl-2′-deoxycytidine, 5-methylcytidine, and N6-methyladenosine) in a single cell. We then further successfully determined DNA and RNA methylation in CTCs from lung cancer patients. Our results demonstrated, for the first time, a significant decrease of DNA methylation (5-methyl-2′-deoxycytidine) and increase of RNA adenine and cytosine methylations (N6-methyladenosine and 5-methylcytidine) in CTCs compared with whole blood cells. The discovery of DNA hypomethylation and RNA hypermethylation in CTCs in the current study together with previous reports of global DNA hypomethylation in tumor tissues suggest that nucleic acid modifications play important roles in the formation and development of cancer cells. This work constitutes the first step for the investigation of DNA and RNA methylation in CTCs, which may facilitate uncovering the metastasis mechanism of cancers in the future.
Co-reporter:Jia-Quan Xu, Huan-Huan Duo, Yu-Ge Zhang, Xin-Wei Zhang, Wei Fang, Yan-Ling Liu, Ai-Guo Shen, Ji-Ming Hu, and Wei-Hua Huang
Analytical Chemistry 2016 Volume 88(Issue 7) pp:3789
Publication Date(Web):March 1, 2016
DOI:10.1021/acs.analchem.5b04810
Biosensors always suffer from passivation that prevents their reutilization. To address this issue, photocatalytically renewable sensors composed of semiconductor photocatalysts and sensing materials have emerged recently. In this work, we developed a robust and versatile method to construct different kinds of renewable biosensors consisting of ZnO nanorods and nanostructured Au. Via a facile and efficient photochemical reduction, various nanostructured Au was obtained successfully on ZnO nanorods. As-prepared sensors concurrently possess excellent sensing capability and desirable photocatalytic cleaning performance. Experimental results demonstrate that dendritic Au/ZnO composite has the strongest surface-enhanced Raman scattering (SERS) enhancement, and dense Au nanoparticles (NPs)/ZnO composite has the highest electrochemical activity, which was successfully used for electrochemical detection of NO release from cells. Furthermore, both of the SERS and electrochemical sensors can be regenerated efficiently for renewable applications via photodegrading adsorbed probe molecules and biomolecules. Our strategy provides an efficient and versatile method to construct various kinds of highly sensitive renewable sensors and might expand the application of the photocatalytically renewable sensor in the biosensing area.
Co-reporter:Huan-Huan Duo, Jia-Quan Xu, Yan-Ling Liu, Zi-He Jin, Xue-Bo Hu, Wei-Hua Huang
Journal of Electroanalytical Chemistry 2016 Volume 781() pp:371-376
Publication Date(Web):15 November 2016
DOI:10.1016/j.jelechem.2016.06.046
•Construction of a renewable electrode by combination of photocatalysts with biosensing materials•Excellent renewable performance in visible light irradiation•Excellent electrochemical performance with low detection limit to nitric oxide•Real-time monitoring of nitric oxide release from living cells and regeneration for reutilizationUltraviolet (UV) light-induced photocatalysts have been utilized to construct renewable electrode to solve the problem of electrode fouling and passivation. However, considering the damages of UV irradiation to environments and biosystems, it is of great significance to develop and apply visible light-induced photocatalysts for biosensing. But for intrinsic visible light photocatalysts, the high electron-hole recombination rate results in the poor photocatalytic performance. Herein, we design the poly(3,4-ethylenedioxythiophene) (PEDOT)-modified TiO2/CdS nanocomposites electrode, which can be efficiently renewed under visible light irradiation for living cell detection. The formation of TiO2/CdS heterojunction structure greatly enhances photocatalysis in visible light region by promoting separation of photogenerated electron–hole pairs. Additionally, the absorption in visible light of PEDOT further accelerates the electrode renewal. PEDOT coating provides a sensitive biosensing interface for electrochemical detection, and meanwhile prevents the cytotoxicity of CdS to cells. This allows electrochemical monitoring of nitric oxide release from living cells and subsequent visible light-induced electrode regeneration, demonstrating great potential of this renewable electrodes in biosensing.
Co-reporter:Yan-Ling Liu;Zi-He Jin;Yan-Hong Liu;Xue-Bo Hu;Yu Qin;Jia-Quan Xu;Dr. Cui-Fang Fan; Wei-Hua Huang
Angewandte Chemie International Edition 2016 Volume 55( Issue 14) pp:4537-4541
Publication Date(Web):
DOI:10.1002/anie.201601276
Abstract
Stretchable electrochemical sensors are conceivably a powerful technique that provides important chemical information to unravel elastic and curvilinear living body. However, no breakthrough was made in stretchable electrochemical device for biological detection. Herein, we synthesized Au nanotubes (NTs) with large aspect ratio to construct an effective stretchable electrochemical sensor. Interlacing network of Au NTs endows the sensor with desirable stability against mechanical deformation, and Au nanostructure provides excellent electrochemical performance and biocompatibility. This allows for the first time, real-time electrochemical monitoring of mechanically sensitive cells on the sensor both in their stretching-free and stretching states as well as sensing of the inner lining of blood vessels. The results demonstrate the great potential of this sensor in electrochemical detection of living body, opening a new window for stretchable electrochemical sensor in biological exploration.
Co-reporter:Yan-Ling Liu;Zi-He Jin;Yan-Hong Liu;Xue-Bo Hu;Yu Qin;Jia-Quan Xu;Dr. Cui-Fang Fan; Wei-Hua Huang
Angewandte Chemie 2016 Volume 128( Issue 14) pp:4613-4617
Publication Date(Web):
DOI:10.1002/ange.201601276
Abstract
Stretchable electrochemical sensors are conceivably a powerful technique that provides important chemical information to unravel elastic and curvilinear living body. However, no breakthrough was made in stretchable electrochemical device for biological detection. Herein, we synthesized Au nanotubes (NTs) with large aspect ratio to construct an effective stretchable electrochemical sensor. Interlacing network of Au NTs endows the sensor with desirable stability against mechanical deformation, and Au nanostructure provides excellent electrochemical performance and biocompatibility. This allows for the first time, real-time electrochemical monitoring of mechanically sensitive cells on the sensor both in their stretching-free and stretching states as well as sensing of the inner lining of blood vessels. The results demonstrate the great potential of this sensor in electrochemical detection of living body, opening a new window for stretchable electrochemical sensor in biological exploration.
Co-reporter:Song-Wei Lv, Ya Liu, Min Xie, Jing Wang, Xue-Wei Yan, Zhen Li, Wei-Guo Dong, and Wei-Hua Huang
ACS Nano 2016 Volume 10(Issue 6) pp:6201
Publication Date(Web):June 14, 2016
DOI:10.1021/acsnano.6b02208
Isolation of single circulating tumor cells (CTCs) from patients is a very challenging technique that may promote the process of individualized antitumor therapies. However, there exist few systems capable of highly efficient capture and release of single CTCs with high viability for downstream analysis and culture. Herein, we designed a near-infrared (NIR) light-responsive substrate for highly efficient immunocapture and biocompatible site-release of CTCs by a combination of the photothermal effect of gold nanorods (GNRs) and a thermoresponsive hydrogel. The substrate was fabricated by imprinting target cancer cells on a GNR-pre-embedded gelatin hydrogel. Micro/nanostructures generated by cell imprinting produce artificial receptors for cancer cells to improve capture efficiency. Temperature-responsive gelatin dissolves rapidly at 37 °C; this allows bulk recovery of captured CTCs at physiological temperature or site-specific release of single CTCs by NIR-mediated photothermal activation of embedded GNRs. Furthermore, the system has been applied to capture, individually release, and genetically analyze CTCs from the whole blood of cancer patients. The multifunctional NIR-responsive platform demonstrates excellent performance in capture and site-release of CTCs with high viability, which provides a robust and versatile means toward individualized antitumor therapies and also shows promising potential for dynamically manipulating cell–substrate interactions in vitro.Keywords: cell imprinting; cell isolation; gold nanorods; NIR photothermal effect; selective site-release; thermoresponsive hydrogel
Co-reporter:Yan-Ling Liu, Xue-Ying Wang, Jia-Quan Xu, Chong Xiao, Yan-Hong Liu, Xin-Wei Zhang, Jun-Tao Liu and Wei-Hua Huang
Chemical Science 2015 vol. 6(Issue 3) pp:1853-1858
Publication Date(Web):28 Jan 2015
DOI:10.1039/C4SC03123G
It is a great challenge to develop electrochemical sensors with superior sensitivity that concurrently possess high biocompatibility for monitoring at the single cell level. Herein we report a novel and reusable biomimetic micro-electrochemical sensor array with nitric oxide (NO) sensing-interface based on metalloporphyrin and 3-aminophenylboronic acid (APBA) co-functionalized reduced graphene oxide (rGO). The assembling of high specificity catalytic but semi-conductive metalloporphyrin with high electric conductive rGO confers the sensor with sub-nanomolar sensitivity. Further coupling with the small cell-adhesive molecule APBA obviously enhances the cytocompatibility of the microsensor without diminishing the sensitivity, while the reversible reactivity between APBA and cell membrane carbohydrates allows practical reusability. The microsensor was successfully used to sensitively monitor, in real-time, the release of NO molecules from human endothelial cells being cultured directly on the sensor. This demonstrates its potential application in the detection of NO with very low bioactive concentrations for the better understanding of its physiological function and for medical tracking of patient states.
Co-reporter:Song-Wei Lv, Jing Wang, Min Xie, Ning-Ning Lu, Zhen Li, Xue-Wei Yan, Si-Liang Cai, Ping-An Zhang, Wei-Guo Dong and Wei-Hua Huang
Chemical Science 2015 vol. 6(Issue 11) pp:6432-6438
Publication Date(Web):30 Jul 2015
DOI:10.1039/C5SC01380A
Isolation, release and culture of rare circulating tumor cells (CTCs) may, if implemented, promote the progress of individualized anti-tumor therapies. To realize the release of CTCs without disruption of their viability for further culture and analysis, we designed an effective photocontrolled CTC capture/release system by combination of photochemistry and immunomagnetic separation. 7-Aminocoumarin was synthesized as the phototrigger to bridge the connection between the anti-EpCAM antibody and the magnetic beads. The coumarin moieties produced cleavage of a C–O bond under both ultraviolet (UV) and near-infrared (NIR) light illumination, breaking the bridge and releasing CTCs from the immunomagnetic beads. Compared with conventional immunomagnetic separation systems, the negative influence of absorbed immunomagnetic beads on further CTCs culture and analysis was effectively eliminated. The system can specifically recognize 102 MCF-7 cells in 1 mL of human whole blood samples with 90% efficiency and 85% purity. Under the irradiation of UV and NIR light, 73 ± 4% and 52 ± 6% of captured cells were released with a viability of 90% and 97%, respectively. Furthermore, this technique has been used to detect CTCs from whole blood of cancer patients with high purity. This study demonstrates that the photochemical-based immunomagnetic separation method for isolating, releasing and culturing CTCs from clinic patients may provide new opportunities for cancer diagnosis and personalized therapy.
Co-reporter:Ning-Ning Lu, Min Xie, Jing Wang, Song-Wei Lv, Jia-Sheng Yi, Wei-Guo Dong, and Wei-Hua Huang
ACS Applied Materials & Interfaces 2015 Volume 7(Issue 16) pp:8817
Publication Date(Web):April 8, 2015
DOI:10.1021/acsami.5b01397
Isolation of rare, pure, and viable circulating tumor cells (CTCs) provides a significant insight in early cancer diagnosis, and release of captured CTCs without damage for ex vivo culture may offer an opportunity for personalized cancer therapy. In this work, we described a biotin-triggered decomposable immunomagnetic system, in which peptide-tagged antibody designed by chemical conjugation was specifically immobilized on engineered protein-coated magnetic beads. The interaction between peptide and engineered protein can be reversibly destroyed by biotin treatment, making capture and release of CTCs possible. Furthermore, the peptide could mediate multiple antibodies’ coimmobilization on engineered protein-coated magnetic beads, by which capture efficiency for CTCs was obviously improved. Quantitative results showed that 70% of captured cells could be released by biotin addition, and 85% of released cells remained viable. In addition, 79% of cancer cells spiked in human whole blood were captured and could also be successfully released for culture. Finally, immunomagnetic beads simultaneously loaded with anti-EpCAM, anti-HER2, and anti-EGFR were successfully applied to isolate and detect CTCs in 17 cancer patients’ peripheral blood samples, and 2–215 CTCs were identified with high purity. These results suggest that our method is reliable and has great potential in CTC detection for CTC-based molecular profiling, diagnosis, and therapy.Keywords: capture; cell viability; circulating tumor cells; peptide; release;
Co-reporter:Xue-Ying Wang, Ying Pei, Min Xie, Zi-He Jin, Ya-Shi Xiao, Yang Wang, Li-Na Zhang, Yan Li and Wei-Hua Huang
Lab on a Chip 2015 vol. 15(Issue 4) pp:1178-1187
Publication Date(Web):22 Dec 2014
DOI:10.1039/C4LC00973H
Reproducing a tumor microenvironment consisting of blood vessels and tumor cells for modeling tumor invasion in vitro is particularly challenging. Here, we report an artificial blood vessel implanted 3D microfluidic system for reproducing transvascular migration of tumor cells. The transparent, porous and elastic artificial blood vessels are obtained by constructing polysaccharide cellulose-based microtubes using a chitosan sacrificial template, and possess excellent cytocompatibility, permeability, and mechanical characteristics. The artificial blood vessels are then fully implanted into the collagen matrix to reconstruct the 3D microsystem for modeling transvascular migration of tumor cells. Well-defined simulated vascular lumens were obtained by proliferation of the human umbilical vein endothelial cells (HUVECs) lining the artificial blood vessels, which enables us to reproduce structures and functions of blood vessels and replicate various hemodynamic parameters. Based on this model, the adhesion and transvascular migration of tumor cells across the artificial blood vessel have been well reproduced.
Co-reporter:Chong Xiao, Yan-Ling Liu, Jia-Quan Xu, Song-Wei Lv, Shan Guo and Wei-Hua Huang
Analyst 2015 vol. 140(Issue 11) pp:3753-3758
Publication Date(Web):12 Jan 2015
DOI:10.1039/C4AN02056A
Here, we report a self-supported nanoporous gold microelectrode decorated with well-dispersed and tiny platinum nanoparticles as an electrochemical nonenzymatic hydrogen peroxide biosensor. Nanoporous gold was fabricated by electrochemical alloying/dealloying and then small-sized platinum nanoparticles were electrodeposited uniformly on them. This novel hybrid nanostructure endows the sensor with high sensitivity and selectivity towards the reduction of hydrogen peroxide with a low detection limit of 0.3 nM. The sensor has been successfully applied for the measurement of H2O2 release from a single isolated human breast cancer cell, demonstrating its great potential for further physiological and pathological applications.
Co-reporter:Jia-Quan Xu;Yan-Ling Liu;Qian Wang;Huan-Huan Duo;Xin-Wei Zhang;Dr. Yu-Tao Li ; Wei-Hua Huang
Angewandte Chemie 2015 Volume 127( Issue 48) pp:14610-14614
Publication Date(Web):
DOI:10.1002/ange.201507354
Abstract
Electrode fouling and passivation is a substantial and inevitable limitation in electrochemical biosensing, and it is a great challenge to efficiently remove the contaminant without changing the surface structure and electrochemical performance. Herein, we propose a versatile and efficient strategy based on photocatalytic cleaning to construct renewable electrochemical sensors for cell analysis. This kind of sensor was fabricated by controllable assembly of reduced graphene oxide (RGO) and TiO2 to form a sandwiching RGO@TiO2 structure, followed by deposition of Au nanoparticles (NPs) onto the RGO shell. The Au NPs-RGO composite shell provides high electrochemical performance. Meanwhile, the encapsulated TiO2 ensures an excellent photocatalytic cleaning property. Application of this renewable microsensor for detection of nitric oxide (NO) release from cells demonstrates the great potential of this strategy in electrode regeneration and biosensing.
Co-reporter:Jia-Quan Xu;Yan-Ling Liu;Qian Wang;Huan-Huan Duo;Xin-Wei Zhang;Dr. Yu-Tao Li ; Wei-Hua Huang
Angewandte Chemie International Edition 2015 Volume 54( Issue 48) pp:14402-14406
Publication Date(Web):
DOI:10.1002/anie.201507354
Abstract
Electrode fouling and passivation is a substantial and inevitable limitation in electrochemical biosensing, and it is a great challenge to efficiently remove the contaminant without changing the surface structure and electrochemical performance. Herein, we propose a versatile and efficient strategy based on photocatalytic cleaning to construct renewable electrochemical sensors for cell analysis. This kind of sensor was fabricated by controllable assembly of reduced graphene oxide (RGO) and TiO2 to form a sandwiching RGO@TiO2 structure, followed by deposition of Au nanoparticles (NPs) onto the RGO shell. The Au NPs-RGO composite shell provides high electrochemical performance. Meanwhile, the encapsulated TiO2 ensures an excellent photocatalytic cleaning property. Application of this renewable microsensor for detection of nitric oxide (NO) release from cells demonstrates the great potential of this strategy in electrode regeneration and biosensing.
Co-reporter:Rong-Rong Xiao, Lei Wang, Lin Zhang, Yu-Ning Liu, Xiao-Lei Yu, and Wei-Hua Huang
Analytical Chemistry 2014 Volume 86(Issue 23) pp:11649
Publication Date(Web):November 10, 2014
DOI:10.1021/ac504159g
Axons are very sensitive to molecular gradients and can discriminate extremely small differences in gradient steepness. Microfluidic devices capable of generating chemical gradients and adjusting their steepness could be used to quantify the sensitivity of axonal response. Here, we present a versatile and robust microfluidic device that can generate substrate-bound molecular gradients with evenly varying steepness on a single chip to precisely quantify axonal response. In this device, two solutions are perfused into a central channel via two inlets while partially flowing into two peripheral channels through interconnecting grooves, which gradually decrease the fluid velocity along the central channel. Molecular gradients with evenly and gradually decreased steepness can therefore be generated with a high resolution that is less than 0.05%/mm. In addition, the overall distribution range and resolution of the gradient steepness can be highly and flexibly controlled by adjusting various parameters of the device. Using this device, we quantified the hippocampal axonal response to substrate-bound laminin and ephrin-A5 gradients with varying steepnesses. Our results provided more detailed information on how and to what extent different steepnesses guide hippocampal neuron development during the initial outgrowth. Furthermore, our results show that axons can sensitively respond to very shallow laminin and ephrin-A5 gradients, which could effectively initiate biased differentiation of hippocampal neurons in the steepness range investigated in this study.
Co-reporter:Min Xie, Ning-Ning Lu, Shi-Bo Cheng, Xue-Ying Wang, Ming Wang, Shan Guo, Cong-Ying Wen, Jiao Hu, Dai-Wen Pang, and Wei-Hua Huang
Analytical Chemistry 2014 Volume 86(Issue 9) pp:4618
Publication Date(Web):April 9, 2014
DOI:10.1021/ac500820p
Early detection and isolation of circulating tumor cells (CTCs) can provide helpful information for diagnosis, and functional readouts of CTCs can give deep insight into tumor biology. In this work, we presented a new strategy for simple isolation and release of CTCs using engineered nanobioprobes. The nanobioprobes were constructed by Ca2+-assisted layer-by-layer assembly of alginate onto the surface of fluorescent-magnetic nanospheres, followed by immobilization of biotin-labeled anti-EpCAM. As-prepared anti-EpCAM-functionalized nanobioprobes were characterized with integrated features of anti-EpCAM-directed specific recognition, fluorescent magnetic-driven cell capture, and EDTA-assisted cell release, which can specifically recognize 102 SK-BR-3 cells spiked in 1 mL of lysed blood or human whole blood samples with 89% and 86% capture efficiency, respectively. Our proof-of-concept experiments demonstrated that 65% of captured SK-BR-3 cells were released after EDTA treatment, and nearly 70% of released SK-BR-3 cells kept their viability, which may facilitate molecular profiling and functional readouts of CTCs.
Co-reporter:Xue-Ying Wang, Zi-He Jin, Bo-Wen Gan, Song-Wei Lv, Min Xie and Wei-Hua Huang
Lab on a Chip 2014 vol. 14(Issue 15) pp:2709-2716
Publication Date(Web):12 May 2014
DOI:10.1039/C4LC00069B
Engineering 3D perfusable vascular networks in vitro and reproducing the physiological environment of blood vessels is very challenging for tissue engineering and investigation of blood vessel function. Here, we engineer interconnected 3D microfluidic vascular networks in hydrogels using molded sodium alginate lattice as sacrificial templates. The sacrificial templates are rapidly replicated in polydimethylsiloxane (PDMS) microfluidic chips via Ca2+-crosslinking and then fully encapsulated in hydrogels. Interconnected channels with well controlled size and morphology are obtained by dissolving the monolayer or multilayer templates with EDTA solution. The human umbilical vein endothelial cells (HUVECs) are cultured on the channel linings and proliferated to form vascular lumens. The strong cell adhesion capability and adaptive response to shear stress demonstrate the excellent cytocompatibility of both the template and template-sacrificing process. Furthermore, the barrier function of the endothelial layer is characterized and the results show that a confluent endothelial monolayer is fully developed. Taken together, we develop a facile and rapid approach to engineer a vascular model that could be potentially used in physiological studies of vascular functions and vascular tissue engineering.
Co-reporter:Yu-Tao Li;Shu-Hui Zhang;Dr. Li Wang;Rong-Rong Xiao;Wei Liu;Xin-Wei Zhang; Zhuan Zhou; Christian Amatore; Wei-Hua Huang
Angewandte Chemie 2014 Volume 126( Issue 46) pp:12664-12668
Publication Date(Web):
DOI:10.1002/ange.201404744
Abstract
Chemical neurotransmission occurs at chemical synapses and endocrine glands, but up to now there was no means for direct monitoring of neurotransmitter exocytosis fluxes and their precise kinetics from inside an individual synapse. The fabrication of a novel finite conical nanoelectrode is reported perfectly suited in size and electrochemical properties for probing amperometrically inside what appears to be single synapses and monitoring individual vesicular exocytotic events in real time. This allowed obtaining direct and important physiological evidences which may yield important and new insights into the nature of synaptic communications.
Co-reporter:Yu-Tao Li;Shu-Hui Zhang;Dr. Li Wang;Rong-Rong Xiao;Wei Liu;Xin-Wei Zhang; Zhuan Zhou; Christian Amatore; Wei-Hua Huang
Angewandte Chemie International Edition 2014 Volume 53( Issue 46) pp:12456-12460
Publication Date(Web):
DOI:10.1002/anie.201404744
Abstract
Chemical neurotransmission occurs at chemical synapses and endocrine glands, but up to now there was no means for direct monitoring of neurotransmitter exocytosis fluxes and their precise kinetics from inside an individual synapse. The fabrication of a novel finite conical nanoelectrode is reported perfectly suited in size and electrochemical properties for probing amperometrically inside what appears to be single synapses and monitoring individual vesicular exocytotic events in real time. This allowed obtaining direct and important physiological evidences which may yield important and new insights into the nature of synaptic communications.
Co-reporter:Jun-Tao Liu;Liang-Sheng Hu;Yan-Ling Liu;Dr. Rong-Sheng Chen;Zhi Cheng;Shi-Jing Chen; Christian Amatore; Wei-Hua Huang; Kai-Fu Huo
Angewandte Chemie International Edition 2014 Volume 53( Issue 10) pp:2643-2647
Publication Date(Web):
DOI:10.1002/anie.201308972
Abstract
Recent biochemical results suggest that auxin (IAA) efflux is mediated by a vesicular cycling mechanism, but no direct detection of vesicular IAA release from single plant cells in real-time has been possible up to now. A TiC@C/Pt-QANFA micro-electrochemical sensor has been developed with high sensitivity in detection of IAA, and it allows real-time monitoring and quantification of the quantal release of auxin from single plant protoplast by exocytosis.
Co-reporter:Jun-Tao Liu;Liang-Sheng Hu;Yan-Ling Liu;Dr. Rong-Sheng Chen;Zhi Cheng;Shi-Jing Chen; Christian Amatore; Wei-Hua Huang; Kai-Fu Huo
Angewandte Chemie 2014 Volume 126( Issue 10) pp:2681-2685
Publication Date(Web):
DOI:10.1002/ange.201308972
Abstract
Recent biochemical results suggest that auxin (IAA) efflux is mediated by a vesicular cycling mechanism, but no direct detection of vesicular IAA release from single plant cells in real-time has been possible up to now. A TiC@C/Pt-QANFA micro-electrochemical sensor has been developed with high sensitivity in detection of IAA, and it allows real-time monitoring and quantification of the quantal release of auxin from single plant protoplast by exocytosis.
Co-reporter:Rong-Rong Xiao, Wen-Juan Zeng, Yu-Tao Li, Wei Zou, Lei Wang, Xue-Fei Pei, Min Xie, and Wei-Hua Huang
Analytical Chemistry 2013 Volume 85(Issue 16) pp:7842
Publication Date(Web):July 18, 2013
DOI:10.1021/ac4022055
Over the past decades, various microfluidic devices have been developed to investigate the role of the molecular gradient in axonal development; however, there are very few devices providing quantitative information about the response of axons to molecular gradients with different slopes. Here, we propose a novel laminar-based microfluidic device enabling simultaneous generation of multiple gradients with gradually changed slope on a single chip. This device, with two asymmetrically designed peripheral channels and opposite flow direction, could generate gradients with gradually changed slope in the center channel, enabling us to investigate simultaneously the response of axons to multiple slope gradients with the same batch of neurons. We quantitatively investigated the response of axon growth rate and growth direction to substrate-bound laminin gradients with different slopes using this single-layer chip. Furthermore, we compartmented this gradient generation chip and a cell culture chip by a porous membrane to investigate quantitatively the response of axon growth rate to the gradient of soluble factor netrin-1. The results suggested that contacting with a molecular gradient would effectively accelerate neurites growth and enhance axonal formation, and the axon guidance ratio obviously increased with the increase of gradient slope in a proper range. The capability of generating a molecular gradient with continuously variable slopes on a single chip would open up opportunities for obtaining quantitative information about the sensitivity of axons and other types of cells in response to gradients of various proteins.
Co-reporter:Qin-Shu Kang, Xiao-Fan Shen, Na-Na Hu, Meng-Jia Hu, Hui Liao, Han-Zhong Wang, Zhi-Ke He and Wei-Hua Huang
Analyst 2013 vol. 138(Issue 9) pp:2613-2619
Publication Date(Web):12 Feb 2013
DOI:10.1039/C3AN36744D
In this work, we demonstrate the immunocapture and on-line fluorescence immunoassay of protein and virus based on porous polymer monoliths (PPM) in microfluidic devices. Poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) [poly(GMA-co-EGDMA)] monoliths were successfully synthesized in the polydimethylsiloxane (PDMS) microfluidic channels by in situ UV-initiated free radical polymerization. After surface modification, PPM provides a high-surface area and specific affinity 3D substrate for immunoassays. Combining with well controlled microfluidic devices, the direct immunoassay of IgG and sandwich immunoassay of inactivated H1N1 influenza virus using 5 μL sample has been accomplished, with detection limits of 4 ng mL−1 and less than 10 pg mL−1, respectively. The enhanced detection sensitivity is due to both high surface area of PPM and flow-through design. The detection time was obviously decreased mainly due to the shortened diffusion distance and improved convective mass transfer inside the monolith, which accelerates the reaction kinetics between antigen and antibody. This work provides a novel microfluidic immunoassay platform with high efficiency thereby enabling fast and sensitive immunoassay.
Co-reporter:Lin-Mei Li, Wei Wang, Shu-Hui Zhang, Shi-Jing Chen, Shi-Shang Guo, Olivier Français, Jie-Ke Cheng, and Wei-Hua Huang
Analytical Chemistry 2011 Volume 83(Issue 24) pp:9524
Publication Date(Web):November 16, 2011
DOI:10.1021/ac202302t
Electrochemical techniques based on ultramicroelectrodes (UMEs) play a significant role in real-time monitoring of chemical messengers’ release from single cells. Conversely, precise monitoring of cells in vitro strongly depends on the adequate construction of cellular physiological microenvironment. In this paper, we developed a multilayer microdevice which integrated high aspect ratio poly(dimethylsiloxane) (PDMS) microfluidic device for long-term automated perfusion culture of cells without shear stress and an independently addressable microelectrodes array (IAMEA) for electrochemical monitoring of the cultured cells in real time. Novel design using high aspect ratio between circular “moat” and ring-shaped micropillar array surrounding cell culture chamber combined with automated “circular-centre” and “bottom-up” perfusion model successfully provided continuous fresh medium and a stable and uniform microenvironment for cells. Two weeks automated culture of human umbilical endothelial cell line (ECV304) and neuronal differentiation of rat pheochromocytoma (PC12) cells have been realized using this device. Furthermore, the quantal release of dopamine from individual PC12 cells during their culture or propagation process was amperometrically monitored in real time. The multifunctional microdevice developed in this paper integrated cellular microenvironment construction and real-time monitoring of cells during their physiological process, and would possibly provide a versatile platform for cell-based biomedical analysis.
Co-reporter:Junhua Wang, Linmei Li, Weihua Huang and Jieke Cheng
Analytical Chemistry 2010 Volume 82(Issue 12) pp:5380
Publication Date(Web):May 14, 2010
DOI:10.1021/ac100007d
Many efforts have been made toward the advancement of capillary electrophoresis chemiluminescence (CE−CL) detection and its applications through continuous development and improvement of interfaces. In this study, a novel rotary cell for CE−CL detection was fabricated and evaluated. A ring-shaped narrow channel with a quartz bottom is made in a cell body to hold CL reactants and act as the reaction chamber. The CE capillary is placed closely to the bottom of the reaction chamber where analyte is deposited into the CL reactants for reactions to occur. Detection is achieved with a photomultiplier tube below the reaction channel. An advantage of the rotary reaction cell is that it renews the reactants at the capillary end as it revolves at a preset speed during experiments. The rotary detection cell presents a new concept to the conventional coaxial-flow configuration by solving the problems of bubble formation and flow blockage that often interrupt the electrophoresis. Two standard proteins, horseradish peroxidase (HRP) and hemoglobin (Hb), were used to evaluate the interface’s performance with luminol/H2O2 CL system. Satisfactory sensitivities (LOD of 0.91 × 10−9 M for HRP, and 4.37 × 10−8 M for Hb at S/N = 3) were obtained in this proof-of-concept experiment. This novel design provides a straightforward and robust interface for CE-CL hyphenation.
Co-reporter:Feng Ai, Hong Chen, Shu-Hui Zhang, Sheng-Yi Liu, Fang Wei, Xu-Yan Dong, Jie-Ke Cheng and Wei-Hua Huang
Analytical Chemistry 2009 Volume 81(Issue 20) pp:8453
Publication Date(Web):September 24, 2009
DOI:10.1021/ac901300b
Oxidative bursts from plants play significant roles in plant disease defense and signal transduction; however, it has not hitherto been investigated on individual living plant cells. In this article, we fabricated a novel sensitive electrochemical sensor based on electrochemical deposition of Pt nanoparticles on the surface of carbon fiber microdisk electrodes via a nanopores containing polymer matrix, Nafion. The numerous hydrophilic nanochannels in the Nafion clusters coated on the electrode surface served as the molecular template for the deposition and dispersion of Pt, which resulted in the uniform construction of small Pt nanoparticles. The novel sensor displayed a high sensitivity for detection of H2O2 with a detection limit of 5.0 × 10−9 M. With the use of this microelectrochemical sensor, the oxidative burst from individual living plant protoplasts have been real-time monitored for the first time. The results showed that oxidative burst from single protoplasts triggered by a pathogen analogue were characterized by quanta release with a large number of “transient oxidative microburst” events, and protoplasts from the transgenic plants biologically displayed better disease-resistance and showed a distinguished elevation and longer-lasting oxidative burst.
Co-reporter:Wei Wang;Shu-Hui Zhang;Lin-Mei Li;Zong-Li Wang
Analytical and Bioanalytical Chemistry 2009 Volume 394( Issue 1) pp:17-32
Publication Date(Web):2009 May
DOI:10.1007/s00216-009-2703-2
Communication between cells by release of specific chemical messengers via exocytosis plays crucial roles in biological process. Electrochemical detection based on ultramicroelectrodes (UMEs) has become one of the most powerful techniques in real-time monitoring of an extremely small number of released molecules during very short time scales, owing to its intrinsic advantages such as fast response, excellent sensitivity, and high spatiotemporal resolution. Great successes have been achieved in the use of UME methods to obtain quantitative and kinetic information about released chemical messengers and to reveal the molecular mechanism in vesicular exocytosis. In this paper, we review recent developments in monitoring exocytosis by use of UMEs-electrochemical-based techniques including electrochemical detection using micrometer and nanometer-sized sensors, scanning electrochemical microscopy (SECM), and UMEs implemented in lab-on-a-chip (LOC) microsystems. These advances are of great significance in obtaining a better understanding of vesicular exocytosis and chemical communications between cells, and will facilitate developments in many fields, including analytical chemistry, biological science, and medicine. Furthermore, future developments in electrochemical probing of exocytosis are also proposed.
Co-reporter:Baoxian Shi;Weihua Huang ;Jieke Cheng
Journal of Separation Science 2008 Volume 31( Issue 6-7) pp:1144-1150
Publication Date(Web):
DOI:10.1002/jssc.200700529
Abstract
A rapid and sensitive method was developed for the analysis of amino acids by microchip electrophoresis with Hg-lamp excitation fluorescence detection. Fluorescein-isothiocyanate (FITC) was chosen to estimate the sensitivity of this system, and the detection limit (S/N = 3) with FITC was 1.7 nM, which showed that the system was sensitive as well as simple. Two derivatizing agents, FITC and ortho-phthalaldehyde (OPA) were employed to label amino acids and were compared in the same fluorescence detection system with an Hg lamp as the excitation source. The separation parameters were optimized in detail. Optimum separation of OPA-labeled amino acids was obtained in less than 200 s with 20 mM borate buffer (pH 9.0) containing 20% acetonitrile and 10 mM β-cyclodextrin. Detection limits for amino acids (alanine (Ala), taurine (Tau), glycine (Gly), glutamic acid (Glu), and aspartic acid (Asp)) of 0.38–1.0 μM were achieved. The method was successfully applied to analysis of amino acids in human vascular endothelial cells (ECV-304). The average amount of amino acids in single ECV-304 cells is estimated to be 5.84 fmol for Ala, 2.78 fmol for Tau, 1.15 fmol for Gly, 3.10 fmol for Glu, and 1.30 fmol for Asp.
Co-reporter:Fuying Du, Weihua Huang, Yinxiang Shi, Zongli Wang, Jieke Cheng
Biosensors and Bioelectronics 2008 Volume 24(Issue 3) pp:415-421
Publication Date(Web):15 November 2008
DOI:10.1016/j.bios.2008.04.020
In this paper, a novel NO electrochemical microsensor, which is fabricated by modifying the surface of a carbon fiber microdisk electrode (CFMDE, diameter: 5–7 μm) with single-walled carbon nanotubes (SWNTs) and Nafion membrane, is reported for the first time. The modification of SWNTs dramatically improves the sensitivity of CFMDEs, and the detection limit for NO is 4.3 nM that is nearly 10 times lower than that from the bare one and lower than most NO electrochemical sensors reported before. The Nafion membrane offers a good barrier to some interferents such as nitrite and ascorbic acid without losing response speed to NO. The sensor has been successfully applied to the measurement of NO release from single isolated human umbilical vein endothelial cells (HUVECs). Real-time amperometric data show that the addition of l-arginine (l-arg) or acetylcholine (ACh) can cause a quick increase in NO production with a maximum concentration of 232 ± 44 nM (n = 5) and 159 ± 29 nM (n = 5), respectively.
Co-reporter:Yan-Ling Liu, Xue-Ying Wang, Jia-Quan Xu, Chong Xiao, Yan-Hong Liu, Xin-Wei Zhang, Jun-Tao Liu and Wei-Hua Huang
Chemical Science (2010-Present) 2015 - vol. 6(Issue 3) pp:NaN1858-1858
Publication Date(Web):2015/01/28
DOI:10.1039/C4SC03123G
It is a great challenge to develop electrochemical sensors with superior sensitivity that concurrently possess high biocompatibility for monitoring at the single cell level. Herein we report a novel and reusable biomimetic micro-electrochemical sensor array with nitric oxide (NO) sensing-interface based on metalloporphyrin and 3-aminophenylboronic acid (APBA) co-functionalized reduced graphene oxide (rGO). The assembling of high specificity catalytic but semi-conductive metalloporphyrin with high electric conductive rGO confers the sensor with sub-nanomolar sensitivity. Further coupling with the small cell-adhesive molecule APBA obviously enhances the cytocompatibility of the microsensor without diminishing the sensitivity, while the reversible reactivity between APBA and cell membrane carbohydrates allows practical reusability. The microsensor was successfully used to sensitively monitor, in real-time, the release of NO molecules from human endothelial cells being cultured directly on the sensor. This demonstrates its potential application in the detection of NO with very low bioactive concentrations for the better understanding of its physiological function and for medical tracking of patient states.
Co-reporter:Song-Wei Lv, Jing Wang, Min Xie, Ning-Ning Lu, Zhen Li, Xue-Wei Yan, Si-Liang Cai, Ping-An Zhang, Wei-Guo Dong and Wei-Hua Huang
Chemical Science (2010-Present) 2015 - vol. 6(Issue 11) pp:NaN6438-6438
Publication Date(Web):2015/07/30
DOI:10.1039/C5SC01380A
Isolation, release and culture of rare circulating tumor cells (CTCs) may, if implemented, promote the progress of individualized anti-tumor therapies. To realize the release of CTCs without disruption of their viability for further culture and analysis, we designed an effective photocontrolled CTC capture/release system by combination of photochemistry and immunomagnetic separation. 7-Aminocoumarin was synthesized as the phototrigger to bridge the connection between the anti-EpCAM antibody and the magnetic beads. The coumarin moieties produced cleavage of a C–O bond under both ultraviolet (UV) and near-infrared (NIR) light illumination, breaking the bridge and releasing CTCs from the immunomagnetic beads. Compared with conventional immunomagnetic separation systems, the negative influence of absorbed immunomagnetic beads on further CTCs culture and analysis was effectively eliminated. The system can specifically recognize 102 MCF-7 cells in 1 mL of human whole blood samples with 90% efficiency and 85% purity. Under the irradiation of UV and NIR light, 73 ± 4% and 52 ± 6% of captured cells were released with a viability of 90% and 97%, respectively. Furthermore, this technique has been used to detect CTCs from whole blood of cancer patients with high purity. This study demonstrates that the photochemical-based immunomagnetic separation method for isolating, releasing and culturing CTCs from clinic patients may provide new opportunities for cancer diagnosis and personalized therapy.
