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1.
Adv Funct Mater ; 32(8)2022 Feb 16.
Artículo en Inglés | MEDLINE | ID: mdl-35603230

RESUMEN

We report innovative scalable, vertical, ultra-sharp nanowire arrays that are individually addressable to enable long-term, native recordings of intracellular potentials. Stable amplitudes of intracellular potentials from 3D tissue-like networks of neurons and cardiomyocytes are obtained. Individual electrical addressability is necessary for high-fidelity intracellular electrophysiological recordings. This study paves the way toward predictive, high-throughput, and low-cost electrophysiological drug screening platforms.

2.
Acc Chem Res ; 51(11): 2756-2763, 2018 11 20.
Artículo en Inglés | MEDLINE | ID: mdl-30339358

RESUMEN

Metal nanoclusters containing a few to several hundred atoms with sizes ranging from sub-nanometer to ∼2 nm occupy an intermediate size regime that bridges larger plasmonic nanoparticles and smaller metal complexes. With strong quantum confinement, metal nanoclusters exhibit molecule-like properties. This Account focuses on noble metal nanoclusters that are synthesized within a single stranded DNA template. Compared to other ligand protected metal nanoclusters, DNA-templated metal nanoclusters manifest intriguing physical and chemical properties that are heavily influenced by the design of DNA templates. For example, DNA-templated silver nanoclusters can show bright fluorescence, tunable emission colors, and enhanced stability by tuning the sequence of the encapsulating DNA template. DNA-templated gold nanoclusters can also serve as excellent cocatalysts, which are integratable with other biocatalysts such as enzymes. In this Account, DNA-templated silver and gold nanoclusters are selected as paradigm systems to showcase their emergent properties and unique applications. We first discuss the DNA-templated silver nanoclusters with a focus on the creation of a complementary palette of emission colors, which has potential applications for multiplex assays. The importance of the DNA template toward enhanced stability of silver nanoclusters is also demonstrated. We then introduce a special class of activable fluorescence probes that are based on the fluorescence turn-on phenomena of DNA-templated silver nanoclusters, which are named nanocluster beacons (NCBs). NCBs have distinct advantages over molecular beacons for nucleic acid detection, and their emission mechanisms are also discussed in detail. We then discuss a universal method of creating novel DNA-silver nanocluster aptamers for protein detection with high specificity. The remainder of the Account is devoted to the DNA-templated gold nanoclusters. We demonstrate that DNA-gold nanoclusters can serve as enhancers for enzymatic reduction of oxygen, which is one of the most important reactions in biofuel cells. Although DNA-templated metal nanoclusters are still in their infancy, we anticipate they will emerge as a new type of functional nanomaterial with wide applications in biology and energy science. Future research will focus on the synthesis of size selected DNA-metal nanoclusters with atomic monodispersity, structural determination of different sized DNA-metal nanoclusters, and establishment of structure-property correlations. Some long-standing mysteries, such as the origin of fluorescence and mechanism for emission color tunability, constitute the central questions regarding the photophysical properties of DNA-metal nanoclusters. On the application side, more studies are required to understand the interaction between nanocluster and biological systems. In the foreseeable future, one can expect that new biosensors, catalysts, and functional devices will be invented based on the intriguing properties of well-designed DNA-metal nanoclusters and their composites. Overall, DNA-metal nanoclusters can add additional spotlights into the highly vibrant field of ligand protected, quantum sized metal nanoclusters.


Asunto(s)
ADN/química , Nanopartículas del Metal/química , Aptámeros de Nucleótidos/química , Fuentes de Energía Bioeléctrica , Técnicas Biosensibles/métodos , Oro/química , Ácidos Nucleicos/análisis , Oxidación-Reducción , Oxígeno/química , Polimorfismo de Nucleótido Simple , Proteínas/análisis , Plata/química
3.
Bioconjug Chem ; 29(8): 2654-2664, 2018 08 15.
Artículo en Inglés | MEDLINE | ID: mdl-29979588

RESUMEN

Protein-ligand conjugations are usually carried out in aqueous media in order to mimic the environment within which the conjugates will be used. In this work, we focus on the conjugation of amphiphilic variants of elastin-like polypeptide (ELP), short elastin (sEL), to poorly water-soluble compounds like OPPVs ( p-phenylenevinylene oligomers), triarylamines, and polypyridine-metal complexes. These conjugations are problematic when carried out in aqueous phase because hydrophobic ligands tend to avoid exposure to water, which in turn causes the ligand to self-aggregate and/or interact noncovalently with hydrophobic regions of the amphiphile. Ultimately, this behavior leads to low conjugation efficiency and contamination with strong noncovalent "conjugates". After exploring the solubility of sEL in various organic solvents, we have established an efficient conjugation methodology for obtaining covalent conjugates virtually free of contaminating noncovalent complexes. When conjugating carboxylated ligands to the amphiphile amines, we demonstrate that even when only one amine (the N-terminus) is present, its derivatization is 98% efficient. When conjugating amine moieties to the amphiphile carboxyls (a problematic configuration), protein multimerization is avoided, 98-100% of the protein is conjugated, and the unreacted ligand is recovered in pure form. Our syntheses occur in "one pot", and our purification procedure is a simple workup utilizing a combination of water and organic solvent extractions. This conjugation methodology might provide a solution to problems arising from solubility mismatch of protein and ligand, and it is likely to be widely applied for modification of recombinant amphiphiles used for drug delivery (PEG-antibodies, polymer-enzymes, food proteins), cell adhesion (collagen, hydrophobins), synthesis of nanostructures (peptides), and engineering of biocompatible optoelectronics (biological polymers), to cite a few.


Asunto(s)
Aminas/química , Elastina/química , Metales/química , Compuestos Orgánicos/química , Polímeros/química , Piridinas/química , Solventes/química , Electroforesis en Gel de Poliacrilamida , Interacciones Hidrofóbicas e Hidrofílicas , Ligandos , Multimerización de Proteína , Espectroscopía de Protones por Resonancia Magnética , Solubilidad , Espectrofotometría Ultravioleta
4.
J Biol Inorg Chem ; 23(7): 1153-1157, 2018 10.
Artículo en Inglés | MEDLINE | ID: mdl-29934674

RESUMEN

An optically active metallo-polymer assembly is demonstrated via conjugation of a genetically engineered elastin-like polypeptide (ELP) and a ruthenium(II) polypyridyl complex. By taking advantage of the phase transition of ELPs in water, photophysical properties of the resultant conjugate are investigated for both phases, below and above the critical transition temperature. Upon coacervation, the luminescence of the metallo-ELP is greatly enhanced as a consequence of local effects on the metal-ligand luminophore. These findings open a possibility to harness the temperature control of stimuli-responsive properties of biopolymers.


Asunto(s)
Biopolímeros/química , Complejos de Coordinación/química , Elastina/química , Luminiscencia , Péptidos/química , Rutenio/química , Complejos de Coordinación/síntesis química , Conformación Molecular , Procesos Fotoquímicos , Temperatura
5.
J Am Chem Soc ; 137(36): 11678-87, 2015 Sep 16.
Artículo en Inglés | MEDLINE | ID: mdl-26288369

RESUMEN

We report the synthesis and characterization of a new DNA-templated gold nanocluster (AuNC) of ∼1 nm in diameter and possessing ∼7 Au atoms. When integrated with bilirubin oxidase (BOD) and single walled carbon nanotubes (SWNTs), the AuNC acts as an enhancer of electron transfer (ET) and lowers the overpotential of electrocatalytic oxygen reduction reaction (ORR) by ∼15 mV as compared to the enzyme alone. In addition, the presence of AuNC causes significant enhancements in the electrocatalytic current densities at the electrode. Control experiments show that such enhancement of ORR by the AuNC is specific to nanoclusters and not to plasmonic gold particles. Rotating ring disk electrode (RRDE) measurements confirm 4e(-) reduction of O2 to H2O with minimal production of H2O2, suggesting that the presence of AuNC does not perturb the mechanism of ORR catalyzed by the enzyme. This unique role of the AuNC as enhancer of ET at the enzyme-electrode interface makes it a potential candidate for the development of cathodes in enzymatic fuel cells, which often suffer from poor electronic communication between the electrode surface and the enzyme active site. Finally, the AuNC displays phosphorescence with large Stokes shift and microsecond lifetime.


Asunto(s)
ADN/química , Enzimas/química , Oro/química , Nanopartículas del Metal/química , Oxígeno/química , Microscopía Electrónica de Transmisión
6.
Phys Chem Chem Phys ; 17(24): 15675-8, 2015 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-25894733

RESUMEN

Water-soluble cationic conjugated poly(phenylene vinylene) (PPV) and cationic fullerene were complexed with negatively charged single stranded DNA and double stranded DNA via electrostatic interactions to achieve photoinduced charge transfer with efficiencies as high as those observed from oppositely charged, cationic PPV and anionic fullerene but with distinctly different quenching mechanisms.


Asunto(s)
ADN/química , Fulerenos/química , Polivinilos/química , Cationes/química , Estructura Molecular , Procesos Fotoquímicos , Electricidad Estática
7.
J Nanosci Nanotechnol ; 14(1): 250-72, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-24730262

RESUMEN

Polythiophene and its derivatives have shown tremendous potential for interfacing electrically conducting polymers with biological applications. These semiconducting organic polymers are relatively soft, conduct electrons and ions, have low cytotoxicity, and can undergo facile chemical modifications. In addition, the reduction in electrical impedance of electrodes coated with polythiophenes may prove to be invaluable for a stable and permanent connection between devices and biological tissues. This review article focuses on the synthesis and some key applications of polythiophenes in multidisciplinary areas at the interface with biology. These polymers have shown tremendous potential in biological applications such as diagnostics, therapy, drug delivery, imaging, implant devices and artificial organs.


Asunto(s)
Electrodos Implantados , Nanocápsulas , Polímeros , Tiofenos , Materiales Biocompatibles/química , Materiales Biocompatibles/uso terapéutico , Conductividad Eléctrica , Nanocápsulas/química , Nanocápsulas/uso terapéutico , Polímeros/química , Polímeros/uso terapéutico , Diseño de Prótesis , Tiofenos/química , Tiofenos/uso terapéutico
8.
J Am Chem Soc ; 134(28): 11550-8, 2012 Jul 18.
Artículo en Inglés | MEDLINE | ID: mdl-22775452

RESUMEN

Rapid and precise screening of small genetic variations, such as single-nucleotide polymorphisms (SNPs), among an individual's genome is still an unmet challenge at point-of-care settings. One crucial step toward this goal is the development of discrimination probes that require no enzymatic reaction and are easy to use. Here we report a new type of fluorescent molecular probe, termed a chameleon NanoCluster Beacon (cNCB), that lights up into different colors upon binding SNP targets. NanoCluster Beacons (NCBs) are collections of a small number of Ag atoms templated on single-stranded DNA that fluoresce strongly when placed in proximity to particular DNA sequences, termed enhancers. Here we show the fluorescence emission color of a NCB can change substantially (a shift of 60-70 nm in the emission maximum) depending upon the alignment between the silver nanocluster and the DNA enhancer sequence. Chameleon NCBs exploit this color shift to directly detect SNPs, based on the fact that different SNPs produce a different alignment between the Ag nanocluster and the enhancer. This SNP detection method has been validated on all single-nucleotide substitution scenarios in three synthetic DNA targets, in six disease-related SNP targets, and in two clinical samples taken from patients with ovarian serous borderline tumors. Samples with single-nucleotide variations can be easily identified by the naked eye under UV excitation, making this method a reliable and low-cost assay with a simple readout format.


Asunto(s)
Fluorescencia , Nanoestructuras , Polimorfismo Genético , Color , Sondas Moleculares
9.
ACS Appl Mater Interfaces ; 14(13): 14871-14886, 2022 Apr 06.
Artículo en Inglés | MEDLINE | ID: mdl-35344326

RESUMEN

Photodynamic therapy (PDT) has been explored as a therapeutic strategy to clear toxic amyloid aggregates involved in neurodegenerative disorders such as Alzheimer's disease. A major limitation of PDT is off-target oxidation, which can be lethal for the surrounding cells. We have shown that a novel class of oligo-p-phenylene ethynylenes (OPEs) exhibit selective binding and fluorescence turn-on in the presence of prefibrillar and fibrillar aggregates of disease-relevant proteins such as amyloid-ß (Aß) and α-synuclein. Concomitant with fluorescence turn-on, OPE also photosensitizes singlet oxygen under illumination through the generation of a triplet state, pointing to the potential application of OPEs as photosensitizers in PDT. Herein, we investigated the photosensitizing activity of an anionic OPE for the photo-oxidation of Aß fibrils and compared its efficacy to the well-known but nonselective photosensitizer methylene blue (MB). Our results show that, while MB photo-oxidized both monomeric and fibrillar conformers of Aß40, OPE oxidized only Aß40 fibrils, targeting two histidine residues on the fibril surface and a methionine residue located in the fibril core. Oxidized fibrils were shorter and more dispersed but retained the characteristic ß-sheet rich fibrillar structure and the ability to seed further fibril growth. Importantly, the oxidized fibrils displayed low toxicity. We have thus discovered a class of novel theranostics for the simultaneous detection and oxidization of amyloid aggregates. Importantly, the selectivity of OPE's photosensitizing activity overcomes the limitation of off-target oxidation of traditional photosensitizers and represents an advancement of PDT as a viable strategy to treat neurodegenerative disorders.


Asunto(s)
Enfermedad de Alzheimer , Péptidos beta-Amiloides , Enfermedad de Alzheimer/metabolismo , Amiloide/química , Péptidos beta-Amiloides/metabolismo , Proteínas Amiloidogénicas , Humanos , Fragmentos de Péptidos/química , Conformación Proteica en Lámina beta
10.
J Am Chem Soc ; 133(31): 11837-9, 2011 Aug 10.
Artículo en Inglés | MEDLINE | ID: mdl-21770404

RESUMEN

DNA-templated silver nanoclusters are promising biological fluorescence probes due to their useful fluorescence properties, including tunability of emission wavelength through DNA template sequence variations. Ag K-edge EXAFS analysis of DNA-templated silver nanoclusters has been used to obtain insight into silver nanocluster bonding, size, and structural correlations to fluorescence. The results indicate the presence of small silver nanoclusters (<30 silver atoms) containing Ag-Ag bonds and Ag-N/O ligations to DNA. The DNA sequence used leads to differences in silver-DNA ligation as well as silver nanocluster size. The results support a model in which cooperative effects of both Ag-DNA ligation and variations in cluster size lead to the tuning of the fluorescence emission of DNA-templated silver nanoclusters.


Asunto(s)
ADN/química , Fluorescencia , Nanopartículas del Metal/química , Plata/química , Secuencia de Bases , Estructura Molecular , Espectroscopía de Absorción de Rayos X
11.
Nano Lett ; 10(8): 3106-10, 2010 Aug 11.
Artículo en Inglés | MEDLINE | ID: mdl-20698624

RESUMEN

DNA-templated silver nanoclusters (DNA/Ag NCs) are an emerging set of fluorophores that are smaller than semiconductor quantum dots and can have better photostability and brightness than commonly used organic dyes. Here we find the red fluorescence of DNA/Ag NCs can be enhanced 500-fold when placed in proximity to guanine-rich DNA sequences. On the basis of this new phenomenon, we have designed a DNA detection probe (NanoCluster Beacon, NCB) that "lights up" upon target binding. Since NCBs do not rely on Forster energy transfer for quenching, they can easily reach high (>100) signal-to-background ratios (S/B ratios) upon target binding. Here, in a separation-free assay, we demonstrate NCB detection of an influenza target with a S/B ratio of 175, a factor of 5 better than a conventional molecular beacon probe. Since the observed fluorescence enhancement is caused by intrinsic nucleobases, our detection technique is simple, inexpensive, and compatible with commercial DNA synthesizers.


Asunto(s)
ADN/química , Sondas Moleculares , Nanoestructuras , Plata/química , Unión Competitiva , Hibridación de Ácido Nucleico
12.
Chem Commun (Camb) ; 57(77): 9922-9925, 2021 Sep 28.
Artículo en Inglés | MEDLINE | ID: mdl-34498621

RESUMEN

We report a critical advance in the generation and characterization of peptoid hetero-oligomers. A library of sub-monomers with amine and carboxylate side-chains are combined in different sequences using microwave-assisted synthesis. Their sequence-structure propensity is confirmed by circular dichroism, and conformer subtypes are enumerated by NMR. Biasing the ψ-angle backbone to trans (180°) in Monte Carlo modelling favors i to i + 3 naphthyl-naphthyl stacking, and matches experimental ensemble distributions. Taken together, high-yield synthesis of heterooligomers and NMR with structure prediction enables rapid determination of sequences that induce secondary structural propensities for predictive design of hydrophilic peptidomimetic foldamers and their future libraries.

13.
Int J Pharm ; 597: 120340, 2021 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-33545284

RESUMEN

Biocompatible nanoparticles composed of poly(lactic-co-glycolic acid) (PLGA) are used as drug and vaccine delivery systems because of their tunability in size and sustained release of cargo molecules. While the use of toxic stabilizers such as polyvinyl alcohol (PVA) limit the utility of PLGA, stabilizer-free PLGA nanoparticles are rarely used because they can be challenging to prepare. Here, we developed a tunable, stabilizer-free PLGA nanoparticle formulation capable of encapsulating plasmid DNA and demonstrated the formation of an elastin-like polymer PLGA hybrid nanoparticle with exceptional stability and biocompatibility. A suite of PLGAs were fabricated using solvent evaporation methods and assessed for particle size and stability in water. We find that under physiological conditions (PBS at 37˚C), the most stable PLGA formulation (P4) was found to contain a greater L:G ratio (65:35), lower MW, and carboxyl terminus. Subsequent experiments determined P4 nanoparticles were as stable as those made with PVA, yet significantly less cytotoxic. Variation in particle size was achieved through altering PLGA stoichiometry while maintaining the ability to encapsulate DNA and were modified with elastin-like polymers for increased immune tolerance. Overall, a useful method for tunable, stabilizer-free PLGA nanoparticle formulation was developed for use in drug and vaccine delivery, and immune targeting.


Asunto(s)
Nanopartículas , Ácido Poliglicólico , Sistemas de Liberación de Medicamentos , Elastina , Ácido Láctico , Tamaño de la Partícula , Copolímero de Ácido Poliláctico-Ácido Poliglicólico
14.
Anal Chem ; 82(1): 136-44, 2010 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-20000585

RESUMEN

No single biomarker can accurately predict disease. An ideal biodetection technology should be capable of the quantitative, reproducible, and sensitive detection of a limited suite of such molecules. To this end, we have developed a multiplex biomarker assay for protective antigen and lethal factor of the Bacillus anthracis lethal toxin using semiconductor quantum dots as the fluorescence reporters on our waveguide-based biosensor platform. The platform is extendable to a wide array of biomarkers, facilitating rapid, quantitative, sensitive, and multiplex detection, better than achievable by conventional immunoassay. Our assay allows for the sensitive (limit of detection 1 pM each), specific (minimal nonspecific binding), and rapid (15 min) detection of these biomarkers in complex biological samples (e.g., serum). To address the issue of reproducibility in measurement and to increase our sample throughput, we have incorporated multichannel waveguides capable of simultaneous multiplex detection of biomarkers in three samples in quadruplicate. In this paper, we present the design, fabrication, and development of multichannel waveguides for the simultaneous detection of lethal factor and protective antigen in serum. Evaluation of the multichannel waveguide shows an excellent concordance with single-channel data and effective, simultaneous, and reproducible measurement of lethal toxins in three samples.


Asunto(s)
Biomarcadores , Inmunohistoquímica/instrumentación , Inmunohistoquímica/métodos , Anticuerpos Antibacterianos , Antígenos Bacterianos , Toxinas Bacterianas , Proteínas Inmovilizadas , Puntos Cuánticos , Reproducibilidad de los Resultados , Sensibilidad y Especificidad
15.
Sensors (Basel) ; 9(7): 5783-809, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-22346727

RESUMEN

Optical phenomena such as fluorescence, phosphorescence, polarization, interference and non-linearity have been extensively used for biosensing applications. Optical waveguides (both planar and fiber-optic) are comprised of a material with high permittivity/high refractive index surrounded on all sides by materials with lower refractive indices, such as a substrate and the media to be sensed. This arrangement allows coupled light to propagate through the high refractive index waveguide by total internal reflection and generates an electromagnetic wave-the evanescent field-whose amplitude decreases exponentially as the distance from the surface increases. Excitation of fluorophores within the evanescent wave allows for sensitive detection while minimizing background fluorescence from complex, "dirty" biological samples. In this review, we will describe the basic principles, advantages and disadvantages of planar optical waveguide-based biodetection technologies. This discussion will include already commercialized technologies (e.g., Corning's EPIC(®) Ô, SRU Biosystems' BIND(™), Zeptosense(®), etc.) and new technologies that are under research and development. We will also review differing assay approaches for the detection of various biomolecules, as well as the thin-film coatings that are often required for waveguide functionalization and effective detection. Finally, we will discuss reverse-symmetry waveguides, resonant waveguide grating sensors and metal-clad leaky waveguides as alternative signal transducers in optical biosensing.

16.
Materials (Basel) ; 12(4)2019 Feb 17.
Artículo en Inglés | MEDLINE | ID: mdl-30781571

RESUMEN

Polystyrene-b-polyethylene glycol (PS-b-PEG) amphiphilic block copolymers featuring a terminal tridentate N,N,N-ligand (terpyridine) were synthesized for the first time through an efficient route. In this approach, telechelic chain-end modified polystyrenes were produced via reversible addition-fragmentation chain-transfer (RAFT) polymerization by using terpyridine trithiocarbonate as the chain-transfer agent, after which the hydrophilic polyethylene glycol (PEG) block was incorporated into the hydrophobic polystyrene (PS) block in high yields via a thiol-ene process. Following metal-coordination with Mn2+, Fe2+, Ni2+, and Zn2+, the resulting metallo-polymers were self-assembled into spherical, vesicular nanostructures, as characterized by dynamic light scattering and transmission electron microscopy (TEM) imaging.

17.
Nucleic Acids Res ; 34(19): e132, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-17023486

RESUMEN

In the use of non-antibody proteins as affinity reagents, diversity has generally been derived from oligonucleotide-encoded random amino acids. Although specific binders of high-affinity have been selected from such libraries, random oligonucleotides often encode stop codons and amino acid combinations that affect protein folding. Recently it has been shown that specific antibody binding loops grafted into heterologous proteins can confer the specific antibody binding activity to the created chimeric protein. In this paper, we examine the use of such antibody binding loops as diversity elements. We first show that we are able to graft a lysozyme-binding antibody loop into green fluorescent protein (GFP), creating a fluorescent protein with lysozyme-binding activity. Subsequently we have developed a PCR method to harvest random binding loops from antibodies and insert them at predefined sites in any protein, using GFP as an example. The majority of such GFP chimeras remain fluorescent, indicating that binding loops do not disrupt folding. This method can be adapted to the creation of other nucleic acid libraries where diversity is flanked by regions of relative sequence conservation, and its availability sets the stage for the use of antibody loop libraries as diversity elements for selection experiments.


Asunto(s)
Regiones Determinantes de Complementariedad/genética , Genes Reporteros , Proteínas Fluorescentes Verdes/genética , Reacción en Cadena de la Polimerasa/métodos , Proteínas Recombinantes de Fusión/inmunología , Diversidad de Anticuerpos , Secuencia de Bases , Clonación Molecular/métodos , Ensayo de Inmunoadsorción Enzimática , Biblioteca de Genes , Proteínas Fluorescentes Verdes/análisis , Humanos , Datos de Secuencia Molecular , Muramidasa/inmunología , Proteínas Recombinantes de Fusión/análisis , Proteínas Recombinantes de Fusión/genética , Análisis de Secuencia de ADN
18.
J Immunol Methods ; 321(1-2): 60-9, 2007 Apr 10.
Artículo en Inglés | MEDLINE | ID: mdl-17336997

RESUMEN

Rodent-borne hantaviruses cause hemorrhagic fever with renal syndrome (HFRS) in the old world and hantavirus cardio-pulmonary syndrome (HCPS) in the new. Most cases of HCPS in North America are caused by Sin Nombre Virus (SNV). Current viral detection technologies depend upon the identification of anti-viral antibodies in patient serum. Detection of viral antigen may facilitate earlier detection of the pathogen. We describe here the characterization of two single-chain Fv antibodies (scFvs), selected from a large naïve phage antibody library, which are capable of identifying the Sin Nombre Virus nucleocapsid protein (SNV-N), with no cross reactivity with the nucleocapsid protein from other hantaviruses. The utility of such selected scFvs was increased by the creation of an scFv-alkaline phosphatase fusion protein which was able to directly detect virally produced material without the need for additional reagents.


Asunto(s)
Anticuerpos Antivirales/inmunología , Síndrome Pulmonar por Hantavirus/inmunología , Región Variable de Inmunoglobulina/inmunología , Proteínas de la Nucleocápside/inmunología , Virus Sin Nombre/inmunología , Fosfatasa Alcalina/genética , Fosfatasa Alcalina/inmunología , Animales , Anticuerpos Antivirales/genética , Afinidad de Anticuerpos , Especificidad de Anticuerpos , Chlorocebus aethiops , Clonación Molecular , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática , Síndrome Pulmonar por Hantavirus/diagnóstico , Región Variable de Inmunoglobulina/genética , Proteínas de la Nucleocápside/análisis , Proteínas de la Nucleocápside/genética , Biblioteca de Péptidos , Proteínas Recombinantes de Fusión/inmunología , Proteínas Recombinantes/inmunología , Virus Sin Nombre/genética , Células Vero
19.
J Inorg Biochem ; 101(11-12): 1692-8, 2007 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-17868890

RESUMEN

Marinobactins A-E are a suite of amphiphilic siderophores which have a common peptidic head group that coordinates Fe(III), and a fatty acid which varies in length and saturation. As a result of the amphiphilic properties of these siderophores it is difficult to study siderophore-mediated uptake of iron, because the amphiphilic siderophores partition indiscriminately in microbial and other membranes. An alternative method to distinguish amphiphilic siderophore partitioning versus siderophore-mediated active uptake for Fe(III)-marinobactin E has been developed. In addition, a new member of the marinobactin family of siderophores is also reported, marinobactin F, which has a C(18) fatty acid with one double bond and which is substantially more hydrophobic that marinobactins A-E.


Asunto(s)
Oligopéptidos/química , Ácidos Palmíticos/química , Sideróforos/química , Membrana Celular/metabolismo , Electroforesis en Gel de Poliacrilamida , Hierro/química , Hierro/metabolismo , Marinobacter/metabolismo , Estructura Molecular , Oligopéptidos/metabolismo , Oligopéptidos/farmacocinética , Ácidos Palmíticos/metabolismo , Ácidos Palmíticos/farmacocinética , Sideróforos/metabolismo , Sideróforos/farmacocinética , Espectrometría de Masa por Ionización de Electrospray
20.
J Biomed Opt ; 21(9): 94003, 2016 09 01.
Artículo en Inglés | MEDLINE | ID: mdl-27685813

RESUMEN

Chromosome ends are shielded from exonucleolytic attack and inappropriate end-joining by terminal structures called telomeres; these structures are potential targets for anticancer drugs. Telomeres are composed of a simple DNA sequence (5?-TTAGGG-3? in humans) repeated more than a thousand times, a short 3? single-stranded overhang, and numerous proteins. Electron microscopy has shown that the 3? overhang pairs with the complementary strand at an internal site creating a small displacement loop and a large double-stranded "t-loop." Our goal is to determine whether all telomeres adopt the t-loop configuration, or whether there are two or more distinct configurations. Progress in optimizing super-resolution (SR) microscopy for this ongoing investigation is reported here. Results suggest that under certain conditions sample preparation procedures may disrupt chromatin by causing loss of nucleosomes. This finding may limit the use of SR microscopy in telomere studies.

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