Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 12 de 12
Filtrar
Más filtros













Base de datos
Intervalo de año de publicación
1.
Biosens Bioelectron ; 169: 112572, 2020 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-32916610

RESUMEN

Convalescent serum with a high abundance of neutralization IgG is a promising therapeutic agent for rescuing COVID-19 patients in the critical stage. Knowing the concentration of SARS-CoV-2 S1-specific IgG is crucial in selecting appropriate convalescent serum donors. Here, we present a portable microfluidic ELISA technology for rapid (15 min), quantitative, and sensitive detection of anti-SARS-CoV-2 S1 IgG in human serum with only 8 µL sample volume. We first identified a humanized monoclonal IgG that has a high binding affinity and a relatively high specificity towards SARS-CoV-2 S1 protein, which can subsequently serve as the calibration standard of anti-SARS-CoV-2 S1 IgG in serological analyses. We then measured the abundance of anti-SARS-CoV-2 S1 IgG in 16 convalescent COVID-19 patients. Due to the availability of the calibration standard and the large dynamic range of our assay, we were able to identify "qualified donors" for convalescent serum therapy with only one fixed dilution factor (200 ×). Finally, we demonstrated that our technology can sensitively detect SARS-CoV-2 antigens (S1 and N proteins) with pg/mL level sensitivities in 40 min. Overall, our technology can greatly facilitate rapid, sensitive, and quantitative analysis of COVID-19 related markers for therapeutic, diagnostic, epidemiologic, and prognostic purposes.


Asunto(s)
Anticuerpos Antivirales/sangre , Betacoronavirus/inmunología , Infecciones por Coronavirus/virología , Ensayo de Inmunoadsorción Enzimática/instrumentación , Inmunoglobulina G/sangre , Técnicas Analíticas Microfluídicas/instrumentación , Neumonía Viral/virología , Adolescente , Adulto , Anticuerpos Antivirales/inmunología , Antígenos Virales/sangre , Antígenos Virales/inmunología , Técnicas Biosensibles/economía , Técnicas Biosensibles/instrumentación , COVID-19 , Infecciones por Coronavirus/terapia , Ensayo de Inmunoadsorción Enzimática/economía , Diseño de Equipo , Humanos , Inmunización Pasiva , Inmunoglobulina G/inmunología , Límite de Detección , Mediciones Luminiscentes/economía , Mediciones Luminiscentes/instrumentación , Técnicas Analíticas Microfluídicas/economía , Persona de Mediana Edad , Pandemias , Neumonía Viral/terapia , SARS-CoV-2 , Factores de Tiempo , Adulto Joven , Sueroterapia para COVID-19
2.
Lab Chip ; 20(3): 634-646, 2020 02 07.
Artículo en Inglés | MEDLINE | ID: mdl-31922156

RESUMEN

The human-derived orthotopic xenograft mouse model is an effective platform for performing in vivo bladder cancer studies to examine tumor development, metastasis, and therapeutic effects of drugs. To date, the surveillance of tumor progression in real time for orthotopic bladder xenografts is highly dependent on semi-quantitative in vivo imaging technologies such as bioluminescence. While these imaging technologies can estimate tumor progression, they are burdened with requirements such as anesthetics, specialized equipment, and genetic modification of the injected cell line. Thus, a convenient and non-invasive technology to quantitatively monitor the growth of bladder cancer in orthotopic xenografts is highly desired. In this work, using a microfluidic chemiluminescent ELISA platform, we have successfully developed a rapid, multiparameter urine-based and non-invasive biomolecular prognostic technology for orthotopic bladder cancer xenografts. This method consists of two steps. First, the concentrations of a panel of four urinary biomarkers are quantified from the urine of mice bearing orthotopic bladder xenografts. Second, machine learning and principal component analysis (PCA) algorithms are applied to analyze the urinary biomarkers, and subsequently, a score is assigned to indicate the tumor growth. With this methodology, we have quantitatively monitored the orthotopic growth of human bladder cancer that was inoculated with low, medium, and high cancer cell numbers. We also employed this method and performed a proof of principle experiment to examine the in vivo therapeutic efficacy of the EGFR inhibitor, dacomitinib.


Asunto(s)
Neoplasias de la Vejiga Urinaria/orina , Animales , Línea Celular Tumoral , Ensayo de Inmunoadsorción Enzimática , Humanos , Dispositivos Laboratorio en un Chip , Mediciones Luminiscentes , Ratones , Vigilancia de la Población , Neoplasias de la Vejiga Urinaria/diagnóstico por imagen
3.
Analyst ; 145(4): 1346-1354, 2020 Feb 17.
Artículo en Inglés | MEDLINE | ID: mdl-31967116

RESUMEN

Rapid and sensitive detection of drugs of abuse plays an important role in monitoring of drug use and treatment compliance. Sweat based drug analysis shows great advantages due to its non-invasive nature. However, most of the related methods developed to date are qualitative, slow, or costly, which significantly hinders their application in field use. Here we report rapid, sensitive, quantitative detection of drugs of abuse in sweat based on capillary arrays combined with competitive enzyme-linked immunosorbent assay. Using four common drugs of abuse, methadone, methamphetamine, amphetamine, and tetrahydrocannabinol, spiked in artificial sweat as a model system, we demonstrate rapid, quantitative, and multiplexed detection of the four drugs in ∼16 minutes with a low sweat volume (∼4 µL per analyte) and a large dynamic range (methadone: 0.0016 ng mL-1-1 ng mL-1; METH: 0.016 ng mL-1-25 ng mL-1; amphetamine: 0.005 ng mL-1-10 ng mL-1; THC: 0.02 ng mL-1-1000 ng mL-1). In addition, we show that the detection range can be tuned for different applications by adjusting the competitors' concentrations. Our work paves a way to develop an autonomous, portable, and cost-effective device for hospital testing, workplace drug-use screening, roadside testing, and patient monitoring in drug rehabilitation centers.


Asunto(s)
Técnicas Biosensibles/métodos , Drogas Ilícitas/análisis , Detección de Abuso de Sustancias/métodos , Sudor/química , Anfetamina/análisis , Técnicas Biosensibles/instrumentación , Dronabinol/análisis , Ensayo de Inmunoadsorción Enzimática , Humanos , Metadona/análisis , Metanfetamina/análisis , Modelos Teóricos , Sensibilidad y Especificidad , Detección de Abuso de Sustancias/instrumentación , Factores de Tiempo
4.
ACS Sens ; 3(11): 2327-2334, 2018 11 26.
Artículo en Inglés | MEDLINE | ID: mdl-30335974

RESUMEN

Follicle stimulating hormone (FSH) plays a critical role in female reproductive development and homeostasis. The blood/serum concentration of FSH is an important marker for reporting multiple endocrinal functions. The standardized method for mouse FSH (mFSH) quantification based on radioimmunoassay (RIA) suffers from long assay time (∼2 days), relatively low sensitivity, larger sample volume (60 µL), and small dynamic range (2-60 ng/mL); thus, it is insufficient for monitoring fast developing events with relatively small mFSH fluctuations (e.g., estrous cycles of mammals). Here, we developed an automated microfluidic chemiluminescent ELISA device along with the disposal sensor array and the corresponding detection protocol for rapid and quantitative analysis of mFSH from mouse tail serum samples. With this technology, highly sensitive quantification of mFSH can be accomplished within 30 min using only 8 µL of the serum sample. It is further shown that our technique is able to generate results comparable to RIA but has a significantly improved dynamic range that covers 0.5-250 ng/mL. The performance of this technology was evaluated with blood samples collected from ovariectomized animals and animals with reimplanted ovarian tissues, which restored ovarian endocrine function and correlated with estrus cycle analysis study.


Asunto(s)
Ensayo de Inmunoadsorción Enzimática/métodos , Ciclo Estral/fisiología , Hormona Folículo Estimulante/sangre , Técnicas Analíticas Microfluídicas/métodos , Animales , Anticuerpos Monoclonales de Origen Murino/inmunología , Biomarcadores/sangre , Biomarcadores/metabolismo , Ensayo de Inmunoadsorción Enzimática/instrumentación , Femenino , Hormona Folículo Estimulante/inmunología , Hormona Folículo Estimulante/metabolismo , Dispositivos Laboratorio en un Chip , Ratones , Técnicas Analíticas Microfluídicas/instrumentación , Ovario/citología , Vagina/citología
5.
Biosens Bioelectron ; 96: 351-357, 2017 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-28525854

RESUMEN

We report an optofluidic catalytic laser for sensitive sulfide ion detection. In the catalytic reaction, horseradish peroxidase (HRP) enzyme is used for catalyzing the non-fluorescent substrate, 10-Acetyl-3,7-dihydroxyphenox-azine (ADHP), to produce highly fluorescent resorufin, which was used as gain medium for lasing. Using sulfide ions as inhibitors, the catalytic reaction slows down, resulting in a delay in the lasing onset time, which is used as the sensing signal. The sensing mechanism of the catalytic laser is theoretically analyzed and the performance is experimentally characterized. Sulfide anion is chosen as a model ion because of its broad adverse impacts on both environment and human health. Due to the optical feedback provided by the laser, the small difference in the sulfide ion concentration can be amplified. Consequently, a detection limit of 10nM is achieved with a dynamic range as large as three orders of magnitude, representing significant improvement over the traditional fluorescence and colorimetric methods. This work will open a door to a new catalytic-laser-based chemical sensing platform for detecting a wide range of species that could inhibit the catalytic reaction.


Asunto(s)
Técnicas Biosensibles/instrumentación , Sulfuros/análisis , Contaminantes Ambientales/análisis , Diseño de Equipo , Peroxidasa de Rábano Silvestre/química , Rayos Láser , Límite de Detección , Dispositivos Ópticos , Oxazinas/química , Espectrometría de Fluorescencia/instrumentación
6.
Analyst ; 142(13): 2378-2385, 2017 Jun 26.
Artículo en Inglés | MEDLINE | ID: mdl-28548141

RESUMEN

Enzyme-linked immunosorbent assay (ELISA) is widely used in medical diagnostics and fundamental biological research due to its high specificity and reproducibility. However, the traditional 96-well-plate based ELISA still suffers from several notable drawbacks, such as long assay time (4-6 hours), burdensome procedures and large sample/reagent volumes (∼100 µl), which significantly limit traditional ELISA's applications in rapid clinical diagnosis and quasi-real-time prognosis of some fast-developing diseases. Here, we developed a user friendly glass capillary array based microfluidic ELISA device. Benefiting from the high surface-to-volume ratio of the capillary and the rapid chemiluminescent photo-imaging method with a commercial camera, our capillary based ELISA device significantly reduced the sample volume to 20 µL and shortened the total assay time to as short as 16 minutes (including detection time), which represent approximately 10-fold and 5-fold reduction in assay time and sample volume, respectively, in comparison with the traditional plate-based method. Furthermore, through the double exposure method, a nearly 10-fold increase in the detection dynamic range was achieved over the traditional well-based ELISA. Our device can be broadly used in rapid biochemical analysis for biomedicine and research/development laboratories.


Asunto(s)
Ensayo de Inmunoadsorción Enzimática , Dispositivos Laboratorio en un Chip , Microfluídica/métodos , Forma MB de la Creatina-Quinasa/análisis , Vidrio , Humanos , Interleucina-6/análisis , Masculino , Reproducibilidad de los Resultados
7.
Anal Chem ; 84(9): 4214-20, 2012 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-22468727

RESUMEN

We proposed and investigated a novel adaptive two-dimensional (2-D) microgas chromatography system, which consists of one 1st-dimensional column, multiple parallel 2nd-dimensional columns, and a decision-making module. The decision-making module, installed between the 1st- and 2nd-dimensional columns, normally comprises an on-column nondestructive vapor detector, a flow routing system, and a computer that monitors the detection signal from the detector and sends out the trigger signal to the flow routing system. During the operation, effluents from the 1st-dimensional column are first detected by the detector and, then, depending on the signal generated by the detector, routed to one of the 2nd-dimensional columns sequentially for further separation. As compared to conventional 2-D GC systems, the proposed adaptive GC scheme has a number of unique and advantageous features. First and foremost, the multiple parallel columns are independent of each other. Therefore, their length, stationary phase, flow rate, and temperature can be optimized for best separation and maximal versatility. In addition, the adaptive GC significantly lowers the thermal modulator modulation frequency and hence power consumption. Finally, it greatly simplifies the postdata analysis process required to reconstruct the 2-D chromatogram. In this paper, the underlying working principle and data analysis of the adaptive GC was first discussed. Then, separation of a mixture of 20 analytes with various volatilities and polarities was demonstrated using an adaptive GC system with a single 2nd-dimensional column. Finally, an adaptive GC system with dual 2nd-dimensional columns was employed, in conjunction with temperature ramping, in a practical application to separate a mixture of plant emitted volatile organic compounds with significantly shortened analysis time.


Asunto(s)
Cromatografía de Gases/instrumentación , Compuestos Orgánicos Volátiles/aislamiento & purificación , Diseño de Equipo , Volatilización
8.
Anal Chem ; 84(7): 3376-81, 2012 Apr 03.
Artículo en Inglés | MEDLINE | ID: mdl-22413933

RESUMEN

We developed novel flow-through surface-enhanced Raman scattering (SERS) platforms using gold nanoparticle (Au-NP) immobilized multihole capillaries for rapid and sensitive vapor detection. The multihole capillaries consisting of thousands of micrometer-sized flow-through channels provide many unique characteristics for vapor detection. Most importantly, its three-dimensional SERS-active micro-/nanostructures make available multilayered assembly of Au-NPs, which greatly increase SERS-active surface area within a focal volume of excitation and collection, thus improving the detection sensitivity. In addition, the multihole capillary's inherent longitudinal channels offer rapid and convenient vapor delivery, yet its micrometer-sized holes increase the interaction between vapor molecules and SERS-active substrate. Experimentally, rapid pyridine vapor detection (within 1 s of exposure) and ultrasensitive 4-nitrophenol vapor detection (at a sub-ppb level) were successfully achieved in open air at room temperature. Such an ultrasensitive SERS platform enabled, for the first time, the investigation of both pyridine and 4-nitrophenol vapor adsorption isotherms at very low concentrations. Type I and type V behaviors of the International Union of Pure and Applied Chemistry isotherm were well observed, respectively.


Asunto(s)
Oro/química , Nanopartículas del Metal/química , Espectrometría Raman/métodos , Nitrofenoles/química , Piridinas/química , Propiedades de Superficie , Volatilización
9.
ACS Nano ; 6(3): 1939-47, 2012 Mar 27.
Artículo en Inglés | MEDLINE | ID: mdl-22385214

RESUMEN

Photosensitizer, protoporphyrin IX (PpIX), was conjugated with Au nanoparticles (Au NPs) of 19, 66, and 106 nm diameter to study the size-dependent enhancement of reactive oxygen species (ROS) formation enabled by Au NPs. The ROS enhancement ratio is determined to be 1:2.56:4.72 in order of increasing Au NP size, in general agreement with theoretically calculated field enhancement to the fourth power. The convergence of the experimental and simulated results suggests that Au NP-enhanced and size-dependent ROS formation can be attributed directly to the localized electromagnetic field as a result of surface plasmonic resonance of Au NPs under light irradiation. In vitro study on the ROS formation enabled by PpIX-conjugated Au NPs in human breast cancer cells (MDA-MB-231) revealed the similar size-dependent enhancement of intracellular ROS formation, while the enhancement greatly depended on cellular uptake of Au NPs. Cellular photodynamic therapy revealed that cell destruction significantly increased in the presence of Au NPs. Compared to the untreated control (0% destruction), 22.6% cell destruction was seen in the PpIX alone group and more than 50% cell destruction was obtained for all PpIX-conjugated Au NPs. The 66 nm Au NPs yielded the highest cell destruction, consistent with the highest cellular uptake and highest ROS formation. Clearly, the complex cellular environment, size-dependent cellular uptake of Au NPs, and ROS generations are vital contributors to the overall cellular PDT efficacy.


Asunto(s)
Oro/química , Nanopartículas del Metal/química , Tamaño de la Partícula , Protoporfirinas/química , Protoporfirinas/farmacología , Especies Reactivas de Oxígeno/metabolismo , Línea Celular Tumoral , Humanos , Cinética , Modelos Moleculares , Conformación Molecular , Fotoquimioterapia , Fármacos Fotosensibilizantes/química , Fármacos Fotosensibilizantes/farmacología , Relación Estructura-Actividad
10.
Analyst ; 136(13): 2811-7, 2011 Jul 07.
Artículo en Inglés | MEDLINE | ID: mdl-21594246

RESUMEN

We report rapid, sensitive, and direct detection of 2,4-dinitrotoluene (DNT) vapor using tailored gold nanoparticles (Au-NPs) as the SERS substrate. The Au-NPs were synthesized using the UV-assisted photo-chemical reduction method and subsequently formed a monolayer on the glass slide through polymer-mediated self-assembly. The SERS substrate such prepared has high SERS enhancement, high affinity towards DNT vapor, and rapid response to the DNT adsorption/desorption. We systematically studied the effect of the Au-NP size and surface density on the SERS performance such as enhancement factor and response time. With the optimized SERS substrate, an enhancement factor over 5.6 × 10(6) was achieved. Furthermore, real-time detection of DNT vapor with only 0.35 second data acquisition time was demonstrated using a 12 mW laser. Compared to previously reported results, we achieved two orders of magnitude reduction in detection time and more than one order of magnitude reduction in excitation laser power. The detection limit is estimated to be 0.4 attogram, which corresponds to a sub-ppb DNT concentration in air. This work will lead to the development of ultra-fast and ultra-sensitive SERS devices for explosive identification and monitoring.

11.
ACS Nano ; 4(10): 5897-904, 2010 Oct 26.
Artículo en Inglés | MEDLINE | ID: mdl-20925347

RESUMEN

We developed highly sensitive and specific nanosensors based on quantum dots (QDs) and DNAzyme for multiplexed detection of heavy metal ions in liquid. The QDs were coated with a thin silica layer for increased stability and higher quantum yield while maintaining a relatively small size for highly efficient energy transfer. The QD-DNAzyme nanosensors were constructed by conjugating quencher-labeled DNAzymes onto the surface of carboxyl-silanized QDs. In the presence of metal ions, the emission is restored due to the cleavage of DNAzymes. The detection could be completed within 25 min with a single laser excitation source. The detection limit of 0.2 and 0.5 nM was experimentally achieved for Pb(2+) and Cu(2+), respectively, which is a 50- and 70-fold improvement over the recent results obtained with dye molecules. Multiplexed detection was also demonstrated using two different colors of QDs, showing negligible cross-talk between the Pb(2+) detection and Cu(2+) detection.


Asunto(s)
Técnicas Biosensibles/métodos , ADN Catalítico/química , Metales Pesados/química , Nanotecnología/métodos , Puntos Cuánticos , Secuencia de Bases , Cobre/química , Transferencia Resonante de Energía de Fluorescencia , Humanos , Iones , Plomo/química , Metales/química , Datos de Secuencia Molecular , Oligonucleótidos/química , Reproducibilidad de los Resultados , Espectrometría de Fluorescencia/métodos
12.
Opt Lett ; 35(4): 466-8, 2010 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-20160786

RESUMEN

We report numerical simulation and hyperspectral Raman imaging of three index-guiding solid-core photonic crystal fibers (PCFs) of different air-cladding microstructures to assess their respective potential for evanescent-field Raman spectroscopy, with an emphasis on achieving surface-enhanced Raman scattering (SERS) over the entire fiber length. Suspended-core PCF consisting of a silica core surrounded by three large air channels conjoined by a thin silica web is the most robust of the three SERS-active PCFs, with a demonstrated detection sensitivity of 1x10(-10) M R6G in an aqueous solution of only approximately 7.3 microL sampling volume.


Asunto(s)
Fotones , Espectrometría Raman/métodos , Aire , Análisis de Elementos Finitos , Microscopía Electrónica de Rastreo , Dióxido de Silicio/química , Propiedades de Superficie
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA