共查询到20条相似文献,搜索用时 31 毫秒
1.
Azzaroni O Mir M Alvarez M Tiefenauer L Knoll W 《Langmuir : the ACS journal of surfaces and colloids》2008,24(6):2878-2883
In the present work, we describe the properties of a bifunctional redox-labeled bioconjugate at electrode surfaces mediating the electron transfer across the electrode-electrolyte interface. We show that the assembly of ferrocene-labeled streptavidin on biotinylated electrodes results in a reproducible unidirectional current flow in the presence of electron donors in solution. Such rectifying films were built up by spontaneous binding of tetrameric streptavidin molecules to biotin centers immobilized on the electrode surface. Due to the high affinity of biotin to streptavidin, such bifunctional films completely bind any biotinylated compounds. The charge transport between donors in solution and the Au electrode is mediated by the ferrocene moieties, allowing us to develop a molecular rectifier. Our experimental results suggest that such redox-labeled proteins with a high binding capacity constitute a promising alternative to organic compounds used in molecular electronics. 相似文献
2.
Highly fluorescent streptavidin-coated CdSe nanoparticles: preparation in water, characterization, and micropatterning 总被引:1,自引:0,他引:1
Bäumle M Stamou D Segura JM Hovius R Vogel H 《Langmuir : the ACS journal of surfaces and colloids》2004,20(10):3828-3831
CdSe quantum dots (QDs) with a high fluorescence quantum yield of 25% and a narrow size distribution were synthesized in a single step in water using glutathione as a stabilizing molecule. The exceptional optical properties enabled for the first time the detection of in-water-prepared single quantum dots at room temperature. For application as fluorescent bioanalytical probes, the QDs were coated with streptavidin. These QDs self-assemble with high contrast on micropatterned biotin while preserving their optical properties and their capability to bind in addition biotinylated molecules, a prerequisite for the development of novel supramolecular structures and bioassays. 相似文献
3.
Synthesis of monodisperse biotinylated p(NIPAAm)-coated iron oxide magnetic nanoparticles and their bioconjugation to streptavidin 总被引:2,自引:0,他引:2
Narain R Gonzales M Hoffman AS Stayton PS Krishnan KM 《Langmuir : the ACS journal of surfaces and colloids》2007,23(11):6299-6304
We describe here the synthesis of 10 nm, monodisperse, iron oxide nanoparticles that we have coated with temperature-sensitive, biotinylated p(NIPAAm) (b-PNIPAAm). The PNIPAAm was prepared by the reversible addition fragmentation chain transfer polymerization (RAFT), and one end was biotinylated with a PEO maleimide-activated biotin to form a stable thioether linkage. The original synthesized iron oxide particles were stabilized with oleic acid. They were dispersed in dioxane, and the oleic acid molecules were then reversibly exchanged with a mixture of PNIPAAm and b-PNIPAAm at 60 degrees C. The b-PNIPAAm-coated magnetic nanoparticles were found to have an average diameter of approximately 15 nm by dynamic light scattering and transmission electron microscopy. The ability of the biotin terminal groups on the b-PNIPAAm-coated nanoparticles to interact with streptavidin was confirmed by fluorescence and surface plasmon resonance. It was found that the b-PNIPAAm-coated iron oxide nanoparticles can still bind with high affinity to streptavidin in solution or when the streptavidin is immobilized on a surface. We have also demonstrated that the binding of the biotin ligands on the surface of the temperature-responsive magnetic nanoparticles to streptavidin can be turned on and off as a function of temperature. 相似文献
4.
Labeling of the cell surface of Escherichia coli was accomplished by expression of a recombinant outer membrane protein, OmpC, in the presence of the unnatural amino acid azidohomoalanine, which acts as a methionine surrogate. The surface-exposed azide moieties of whole cells were biotinylated via Cu(1)-catalyzed [3+2] azide-alkyne cycloaddition. The specificity of labeling of both wild-type OmpC and a mutant containing additional methionine sites for azidohomoalanine incorporation was confirmed by Western blotting. Flow cytometry was performed to examine the specificity of the labeling. Cells that express the mutant form of OmpC in the presence of azidohomoalanine, which were biotinylated and stained with fluorescent avidin, exhibit a mean fluorescence 10-fold higher than the background. Incorporation of an unnatural amino acid can thus be determined on a single-cell basis. 相似文献
5.
We investigated fluorescence enhancements and lifetime reductions of Cy5 probe molecules at various distances from the deposited silver island film surface using single molecule spectroscopic methods. The proximity of fluorophore molecules to the surface was controlled by alternating layers of biotinylated bovine serum albumin (BSA-biotin) and avidin, followed by binding of Cy5-labeled oligonucleotides to the top of a BSA-biotin layer structure. We observed dramatically varied brightness of fluorophores with distances from metal structures as well with reduced blinking in the presence of silver island films. In addition, distributions of fluorescence lifetimes and apparent emission intensities from individual molecules indicate an inhomogeneous nature of local matrix surface near metallic nanostructures. These studies illustrate the exclusive information that is otherwise hidden in ensemble measurements. 相似文献
6.
Vânia F. Pais Dr. Hamdy S. El‐Sheshtawy Prof. Dr. Rosario Fernández Prof. Dr. José M. Lassaletta Dr. Abel Ros Prof. Dr. Uwe Pischel 《Chemistry (Weinheim an der Bergstrasse, Germany)》2013,19(21):6650-6661
A series of nine borylated arylisoquinolines has been prepared with systematic variation in their electronic properties and their photophysical properties were investigated. The color of their fluorescence can be finely tuned by changing the properties of the aryl moiety, which is involved in internal‐charge‐transfer processes. For example, methoxy‐substituted compound 5 showed an intense green emission, whereas dimethylamino‐substituted compound 6 showed an orange‐red emission. These new fluorophores were tested for their potential as molecular switches with external ionic stimuli, such as protons and fluoride ions. On the one hand, protonation of the isoquinoline moiety led to fluorescence enhancement for compounds that showed weak charge transfer and fluorescence quenching for compounds that showed strong charge transfer. On the other hand, the formation of ate complexes with fluoride led to strong fluorescence quenching in all of the investigated cases. 相似文献
7.
Functional surfaces and especially the control of surface properties depending on external parameters such as light illumination
have gained increasing importance in the last few years. We present the characterization of polymers from the cycloolefin
(co)polymer class (COC/COP) functionalized with an aminosilane as a basis for the further immobilization of compounds. In
a first step, an assay using AlexaFluor?647 fluorescent dye was used to assess surface homogeneity and reproducibility. A
coefficient of variation of less than 15% for dot-to-dot and less than 25% for chip-to-chip could be achieved. The same amino-functionalized
surfaces were then used to immobilize a biotinylated photolabile linker compound, binding AlexaFluor?647-labeled streptavidin.
The linker was photocleaved with high efficiency at λ = 365 nm and P = 0.15 mW/cm2. Fluorescence measurements show that polymers of the COC/COP class can be used as versatile surfaces for the photoinduced
release of compounds immobilized via photolabile linkers. 相似文献
8.
Sulfated glycosaminoglycans were labeled with biotin to study their interaction with cells in culture. Thus, heparin, heparan sulfate, chondroitin 4-sulfate, chondroitin 6-sulfate and dermatan sulfate were labeled using biotin-hydrazide, under different conditions. The structural characteristics of the biotinylated products were determined by chemical (molar ratios of hexosamine, uronic acid, sulfate and biotin) and enzymatic methods (susceptibility to degradation by chondroitinases and heparitinases). The binding of biotinylated glycosaminoglycans was investigated both in endothelial and smooth muscle cells in culture, using a novel time resolved fluorometric method based on interaction of europium-labeled streptavidin with the biotin covalently linked to the compounds. The interactions of glycosaminoglycans were saturable and number of binding sites could be obtained for each individual compound. The apparent dissociation constant varied among the different glycosaminoglycans and between the two cell lines. The interactions of the biotinylated glycosaminoglycans with the cells were also evaluated using confocal microscopy. We propose a convenient and reliable method for the preparation of biotinylated glycosaminoglycans, as well as a sensitive non-competitive fluorescence-based assay for studies of the interactions and binding of these compounds to cells in culture. 相似文献
9.
Qi K Ma Q Remsen EE Clark CG Wooley KL 《Journal of the American Chemical Society》2004,126(21):6599-6607
Shell cross-linked nanoparticles (SCKs) presenting surface- and bioavailable biotin functional groups were synthesized via a mixed micelle methodology, whereby co-micellization of chain terminal biotinylated poly(acrylic acid)-b-poly(methyl acrylate) (PAA-b-PMA) and nonbiotinylated PAA-b-PMA were cross-linked in an intramicellar fashion within the shell layer of the mixed micelles, between the carboxylic acid groups of PAA and the amine functionalities of 2,2'-(ethylenedioxy)diethylamine. The hydrodynamic diameters (D(h)) of the micelles and the SCKs with different biotinylated block copolymer contents were determined by dynamic light scattering (DLS), and the dimensions of the SCKs were characterized with tapping-mode atomic force microscopy (AFM) and transmission electron microscopy (TEM). The amount of surface-available biotin was tuned by varying the stoichiometric ratio of the biotinylated PAA-b-PMA versus the nonbiotinylated PAA-b-PMA, as demonstrated with solution-state, binding interaction analyses, an avidin/HABA (avidin/4'-hydroxyazobenzene-2-carboxylic acid) competitive binding assay, and fluorescence correlation spectroscopy (FCS). The avidin/HABA assay found the amount of available biotin at the surface of the biotinylated SCK nanoparticles to increase with increasing biotin-terminated block copolymer incorporation, but to be less than 25% of the theoretical value. FCS measurements showed the same trend. 相似文献
10.
Matsumoto T Tanaka T Kondo A 《Langmuir : the ACS journal of surfaces and colloids》2012,28(7):3553-3557
A microparticle surface was designed by the unique method incorporating streptavidin-biotin affinity and sortase A (SrtA)-catalyzed transpeptidation. Leucine-proline-glutamate-threonine-glycine-tagged streptavidin (Stav-LPETG)was immobilized on the surface using streptavidin-biotin affinity, and GGGGG-tagged red fluorescent protein (Gly5-RFP) was conjugated with SrtA. Biotinylated fluorescein isothiocyanate (biotin-FITC) was then bound to residual biotin-binding sites in Stav-LPETG. The resulting particles had RFP and FITC immobilized on the surface via Stav-LPETG, and RFP- and FITC-associated fluorescence was observed using fluorescence microscopy. Finally, GGG-tagged glucose oxidase and biotinylated horseradish peroxidase were immobilized on the microparticle surface, resulting in a functional particle capable of detecting glucose. This particle can be repeatedly used and is more sensitive in detecting glucose than particles prepared using chemical modification. Our method provides a simple strategy for site-specific coimmobilization on molecular surfaces and expands the use of protein hybrid devices. 相似文献
11.
Narain R Housni A Gody G Boullanger P Charreyre MT Delair T 《Langmuir : the ACS journal of surfaces and colloids》2007,23(26):12835-12841
We report here the preparation of novel biotinylated glyconanoparticles from well-defined biotinylated glycopolymers and poly(N-isopropylacrylamide) (PNIPAAm) synthesized via the reversible addition fragmentation chain transfer (RAFT) polymerization process. The in situ reduction of the biotinylated glycopolymers, PNIPAAm, poly(ethylene glycol), and HAuCl4 via a photochemical process resulted in the formation of biotinylated gold nanoparticles. The multifunctional biotinylated glyconanoparticles were then evaluated for their bioconjugation toward streptavidin using UV-vis spectroscopy and surface plasmon resonance (SPR). The biotinylated nanoparticles underwent aggregation in the presence of streptavidin as revealed by spectrophotometry, which indicates the accessibility of the biotin for conjugation. These results were further confirmed by surface plasmon resonance even in the case of surface-immobilized streptavidin. 相似文献
12.
The process of surface functionalization involving silanization, biotinylation and streptavidin bonding as platform for biospecific ligand immobilization was optimized for thin film polyimide spin-coated silicon wafers, of which the polyimide film serves as a wave guiding layer in evanescent wave photonic biosensors. This type of optical sensors make great demands on the materials involved as well as on the layer properties, such as the optical quality, the layer thickness and the surface roughness. In this work we realized the binding of a 3-mercaptopropyl trimethoxysilane on an oxygen plasma activated polyimide surface followed by subsequent derivatization of the reactive thiol groups with maleimide-PEG2-biotin and immobilization of streptavidin. The progress of the functionalization was monitored by using different fluorescence labels for optimization of the chemical derivatization steps. Further, X-ray photoelectron spectroscopy and atomic force microscopy were utilized for the characterization of the modified surface. These established analytical methods allowed to derive information like chemical composition of the surface, surface coverage with immobilized streptavidin, as well as parameters of the surface roughness. The proposed functionalization protocol furnished a surface density of 144 fmol mm−2 streptavidin with good reproducibility (13.9% RSD, n = 10) and without inflicted damage to the surface. This surface modification was applied to polyimide based Mach-Zehnder interferometer sensors to realize a real-time measurement of streptavidin binding validating the functionality of the MZI biosensor. Subsequently, this streptavidin surface was employed to immobilize biotinylated single-stranded DNA and utilized for monitoring of selective DNA hybridization. These proved the usability of polyimide based evanescent photonic devices for biosensing application. 相似文献
13.
A series of methano-C60 adducts bearing up to six electron donating N,N-dimethylaniline units (denoted as D compounds), along with their analogues without the dimethylamino groups as references (R compounds), were synthesized. The redox properties of the D compounds in solutions were evaluated spectroscopically in reference to the R compounds. According to UV/vis absorption results, there are obviously ground-state intramolecular charge-transfer complexes in the D series, and the charge-transfer effects apparently become saturated with only two donor units in the molecule. The photoinduced intramolecular electron-transfer properties of the D compounds were investigated via fluorescence measurements. The emission from intramolecular exciplexes can be found only in the D molecule with two electron donor units. Throughout the D series, the fluorescence properties are highly sensitive to the solvent polarity, with the emission completely quenched for all of the molecules in a polar solvent like methylene chloride. Mechanistic implications of the results are discussed. 相似文献
14.
Kessel L Kalinin S Nagaraj RH Larsen M Johansson LB 《Photochemistry and photobiology》2002,76(5):549-554
The intrinsic fluorescence from the human lens on excitation in the UV region, referred to as blue lens autofluorescence, increases with age or in the presence of diabetes. The present study reveals that the relative contribution of compounds responsible for the blue autofluorescence appears to be a constant with age. Three potential candidates for the blue fluorescence were also studied with respect to fluorescence spectroscopic properties. These were argpyrimidine and pentosidine, both advanced glycation end products, and 3-hydroxykynurenine (3-OH-kynurenine), a photooxidative derivative of tryptophan. It was shown that the spectral properties of argpyrimidine and pentosidine are compatible with the observed blue fluorescence of the human lens, whereas the fluorescence from 3-OH-kynurenine is negligible. 相似文献
15.
Here we report the substitution effects of N-alkyl and N,N-dialkyl carboxamide groups on the fluorescence properties of polycyclic aromatic hydrocarbon chromophores, so as to control their fluorescence properties. The fluorescence properties of compounds obtained using solvents with different polarities showed very little change, indicating that the modified compounds do not form charge transfer states. TD-DFT calculations and measurements performed at low temperature (78 K) and in viscous solvents revealed that the N-alkyl and N,N-dialkyl carboxamide groups tend to reduce the contributions from intersystem crossing and increase those from internal conversion. Considering that the fluorescence mechanism of low-fluorescence carbonyl compounds such as aldehyde and ketone is dominated by intersystem crossing and that of high-luminescence carbonyl compounds such as carboxylic acid and ester is dominated by a radiative process, it can be said that the photophysical process of N-alkyl and N,N-dialkyl carboxamides is novel. In addition, the calculation results for excited states indicated that such contributions can be controlled by selecting the appropriate polycyclic aromatic hydrocarbon or amide structure, in addition to solvent viscosity and temperature. 相似文献
16.
Serena Laschi Ilaria Palchetti Giovanna Marrazza Marco Mascini 《Bioelectrochemistry (Amsterdam, Netherlands)》2009,76(1-2):214
In the present study, we investigated the properties of PNA and LNA capture probes in the development of an electrochemical hybridization assay. Streptavidin-coated paramagnetic micro-beads were used as a solid phase to immobilize biotinylated DNA, PNA and LNA capture probes, respectively. The target sequence was then recognized via hybridization with the capture probe. After labeling the biotinylated hybrid with a streptavidin–enzyme conjugate, the electrochemical detection of the enzymatic product was performed onto the surface of a disposable electrode. The assay was applied to the analytical detection of biotinylated DNA as well as RNA sequences. Detection limits, calculated considering the slope of the linear portion of the calibration curve in the range 0–2 nM were found to be 152, 118 and 91 pM, coupled with a reproducibility of the analysis equal to 5, 9 and 6%, calculated as RSD%, for DNA, PNA and LNA probes respectively, using the DNA target. In the case of the RNA target, the detection limits were found to be 51, 60 and 78 pM for DNA, PNA and LNA probes respectively. 相似文献
17.
We report a biosensing method that is based on magnetically immobilized functional liposomes. The vesicles encapsulate magnetic nanoparticles (MNP) and enzymatic sensing reagents. Magnetic attraction between MNP and external magnets first immobilizes liposomes onto the surface of a coverglass. With the assistance from α-hemolysin (aHL), translocations of analytes through a lipid membrane trigger intravesicular enzymatic reactions. After 90 s incubation, the product from the sensing reactions, resorufin, was probed by laser-induced fluorescence. Detection of two analytes, glucose and ethanol, was demonstrated using two types of functional vesicles. The effects of MNP-containing vesicles and biotinylated vesicles on aHL's translocations of analytes were also compared. Unlike biotinylated lipids, MNP facilitate immobilization of liposomes without compromising the integrity of membrane and pore-forming activity of aHL. 相似文献
18.
《Analytical letters》2012,45(14):2341-2349
A simple, label-free fluorescence method was developed for the sensitive determination of lead(II) using a nitrocellulose membrane biosensor. The surface of the nitrocellulose membrane was modified by glutaraldehyde to conjugate streptavidin, followed by the immobilization of a DNA probe via a biotin modifier. The biotinylated DNA probe can fold into a G-quadruplex structure in the presence of potassium ion that selectively binds to N-methyl mesoporphyrin IX and yields a strong fluorescence signal. The presence of lead(II) can induce a conformational change of the G-quadruplex to a more compact structure, which results in the release of potassium ion and N-methyl mesoporphyrin IX with a concomitant reduction of the fluorescence signal. The biosensor displayed a detection limit as low as 10 nM with excellent selectivity for lead(II) over other metal ions. The developed biosensor was employed for the determination of lead(II) in spiked river water. 相似文献
19.
Ornelas C Lodescar R Durandin A Canary JW Pennell R Liebes LF Weck M 《Chemistry (Weinheim an der Bergstrasse, Germany)》2011,17(13):3619-3629
Cyanine dyes are known for their fluorescence in the near-IR (NIR) region, which is desirable for biological applications. We report the synthesis of a series of aminocyanine dyes containing terminal functional groups such as acid, azide, and cyclooctyne groups for further functionalization through, for example, click chemistry. These aminocyanine dyes can be attached to polyfunctional dendrons by copper-catalyzed azide alkyne cycloaddition (CuAAC), strain-promoted azide alkyne cycloaddition (SPAAC), peptide coupling, or direct S(NR)1 reactions. The resulting dendron-dye conjugates were obtained in high yields and displayed high chemical stability and photostability. The optical properties of the new compounds were studied by UV/Vis and fluorescence spectroscopy. All compounds show large Stokes shifts and strong fluorescence in the NIR region with high quantum yields, which are optimal properties for in vivo optical imaging. 相似文献
20.
Sayed M. Saleh Rainer Müller Heike S. Mader Axel Duerkop Otto S. Wolfbeis 《Analytical and bioanalytical chemistry》2010,398(4):1615-1623
Fluorescent silica nanoparticles (SiNPs) were prepared by covalent attachment of fluorophores to the amino-modified surface
of SiNPs with a typical diameter of 15 nm. The SiNPs are intended for use in novel kinds of fluorescence resonance energy
transfer (FRET)-based affinity assays at the interface between nanoparticle and sample solution. Various labels were employed
to obtain a complete set of colored SiNPs, with excitation maxima ranging from 337 to 659 nm and emission maxima ranging from
436 nm to the near infrared (710 nm). The nanoparticles were characterized in terms of size and composition using transmission
electron microscopy, thermogravimetry, elemental analysis, and dynamic light scattering. The surface of the fluorescent SiNPs
was biotinylated, and binding of labeled avidin to the surface was studied via FRET in two model cases. In the first, FRET
occurs from the biotinylated fluorescent SiNP (the donor) to the labeled avidin (the acceptor). In the second, FRET occurs
in the other direction. Aside from its use in the biotin–avidin system, such SiNPs also are believed to be generally useful
fluorescent markers in various kinds of FRET assays, not the least because the fluorophore is located on the surface of the
SiNPs (and thus always much closer to the second fluorophore) rather than being doped deep in its interior. 相似文献