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1.
Proc Natl Acad Sci U S A ; 111(21): 7588-93, 2014 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-24821774

RESUMO

Droplet interface bilayers are versatile model membranes useful for synthetic biology and biosensing; however, to date they have always been confined to fluid reservoirs. Here, we demonstrate that when two or more water droplets collide on an oil-infused substrate, they exhibit noncoalescence due to the formation of a thin oil film that gets squeezed between the droplets from the bottom up. We show that when phospholipids are included in the water droplets, a stable droplet interface bilayer forms between the noncoalescing water droplets. As with traditional oil-submerged droplet interface bilayers, we were able to characterize ion channel transport by incorporating peptides into each droplet. Our findings reveal that droplet interface bilayers can function in ambient environments, which could potentially enable biosensing of airborne matter.


Assuntos
Técnicas Biossensoriais/métodos , Interações Hidrofóbicas e Hidrofílicas , Canais Iônicos/metabolismo , Bicamadas Lipídicas/química , Água/química , Transporte Biológico/fisiologia
2.
Lab Chip ; 7(9): 1184-91, 2007 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17713618

RESUMO

A nanomechanical transducer is developed to detect and screen endocrine disrupting chemicals (EDCs) combining fluidic sample injection and delivery with bioreceptor protein functionalized microcantilevers (MCs). The adverse affects of EDCs on the endocrine system of humans, livestock, and wildlife provides strong motivation for advances in analytical detection and monitoring techniques. The combination of protein receptors, which include estrogen receptor alpha (ER-alpha) and estrogen receptor beta (ER-beta), as well as monoclonal antibodies (Ab), with MC systems employing modified nanostructured surfaces provides for excellent nanomechanical response sensitivity and the inherent selectivity of biospecific receptor-EDC interactions. The observed ranking of binding interaction of the tested EDCs with ER-beta is diethylstilbestrol (DES) > 17-beta-estradiol > 17-alpha-estradiol > 2-OH-estrone > bisphenol A > p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) with measurements exhibiting intra-day RSDs of about 3%. A comparison of responses of three EDCs, which include 17-beta-estradiol, 17-alpha-estradiol, and 2-OH-estrone, with ER-beta and ER-alpha illustrates which estrogen receptor subtype provides the greatest sensitivity. Antibodies specific to a particular EDC can also be used for analyte specific screening. Calibration plots for a MC functionalized with anti-17-beta-estradiol Ab show responses in the range of 1 x 10(-11) through 1 x 10(-7) M for 17-beta-estradiol with a linear portion extending over two orders of magnitude in concentration.


Assuntos
Técnicas Biossensoriais/instrumentação , Disruptores Endócrinos/análise , Poluentes Ambientais/análise , Análise de Injeção de Fluxo/instrumentação , Nanotecnologia/instrumentação , Transdutores , Técnicas Biossensoriais/métodos , Desenho de Equipamento , Análise de Falha de Equipamento , Análise de Injeção de Fluxo/métodos , Miniaturização , Nanotecnologia/métodos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
3.
Anal Chem ; 79(1): 364-70, 2007 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-17194162

RESUMO

The very simple coupling of a standard, packed-column gas chromatograph with a microcantilever array (MCA) is demonstrated for enhanced selectivity and potential analyte identification in the analysis of volatile organic compounds (VOCs). The cantilevers in MCAs are differentially coated on one side with responsive phases (RPs) and produce bending responses of the cantilevers due to analyte-induced surface stresses. Generally, individual components are difficult to elucidate when introduced to MCA systems as mixtures, although pattern recognition techniques are helpful in identifying single components, binary mixtures, or composite responses of distinct mixtures (e.g., fragrances). In the present work, simple test VOC mixtures composed of acetone, ethanol, and trichloroethylene (TCE) in pentane and methanol and acetonitrile in pentane are first separated using a standard gas chromatograph and then introduced into a MCA flow cell. Significant amounts of response diversity to the analytes in the mixtures are demonstrated across the RP-coated cantilevers of the array. Principal component analysis is used to demonstrate that only three components of a four-component VOC mixture could be identified without mixture separation. Calibration studies are performed, demonstrating a good linear response over 2 orders of magnitude for each component in the primary study mixture. Studies of operational parameters including column temperature, column flow rate, and array cell temperature are conducted. Reproducibility studies of VOC peak areas and peak heights are also carried out showing RSDs of less than 4 and 3%, respectively, for intra-assay studies. Of practical significance is the facile manner by which the hyphenation of a mature separation technique and the burgeoning sensing approach is accomplished, and the potential to use pattern recognition techniques with MCAs as a new type of detector for chromatography with analyte-identifying capabilities.


Assuntos
Técnicas Biossensoriais/métodos , Cromatografia Gasosa/métodos , Compostos Orgânicos/análise , Pentanos/análise , Pentanos/química , Acetona/análise , Acetonitrilas/análise , Etanol/análise , Metanol/análise , Análise de Componente Principal , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Tricloroetileno/análise , Volatilização
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