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1.
Adv Mater Interfaces ; 3(23)2016 Dec 07.
Article in English | MEDLINE | ID: mdl-28184332

ABSTRACT

A durable and flexible silicone-based backcoating (halogen free) is applied to the backside of an otherwise smoldering-prone and flammable fabric. When exposed to fire, cyclic siloxanes (produced by thermal decomposition of the backcoating) diffuse through the fabric in the gas phase. The following oxidation of the cyclic siloxanes forms a highly conformal and thermally stable coating that fully embeds all individual fibers and shields them from heat and oxidation. As a result, the combustion of the fabric is prevented. This is a novel fire retardant mechanism that discloses a powerful approach towards textiles and multifunctional flexible materials with combined smoldering/flaming ignition resistance and fire-barrier properties.

2.
J Occup Environ Hyg ; 13(4): 303-13, 2016.
Article in English | MEDLINE | ID: mdl-26647208

ABSTRACT

Certain engineered nanoparticles (ENP) reduce the flammability of components used in soft furnishings (mattresses and upholstered furniture). However, because of the ENP's small size and ability to interact with biological molecules, these fire retardant ENPs may pose a health and environmental risks, if they are released sometime during the life cycle of the soft furnishing. Quantifying the released amount of these ENPs under normal end-use circumstances provides a basis for assessing their potential health and environmental impact. In this article, we report on efforts to identify suitable methodologies for quantifying the release of carbon nanofibers, carbon nanotubes, and sodium montmorillonites from coatings applied to the surfaces of barrier fabric and polyurethane foam. The ENPs released in simulated chewing and mechanical stressing experiments were collected in aqueous solution and quantified using Ultraviolet-Visible and inductively coupled plasma-optical emission spectroscopy. The microstructures of the released ENPs were characterized using scanning electron microscopy. The reported methodology and results provide important milestones to estimate the impact and toxicity of the ENP release during the life cycle of the nanocomposites. To our knowledge, this is the first study of ENP release from the soft furnishing coating, something that can be important application area for fire safety.


Subject(s)
Flame Retardants , Nanocomposites , Nanoparticles/analysis , Stress, Mechanical , Bentonite/analysis , Interior Design and Furnishings , Microscopy, Electron, Scanning , Nanoparticles/ultrastructure , Nanotubes, Carbon/analysis , Polymers/analysis , Polyurethanes , Spectrophotometry
3.
Biosens Bioelectron ; 76: 2-19, 2016 Feb 15.
Article in English | MEDLINE | ID: mdl-26139320

ABSTRACT

It has been more than two decades since aptamer and the systematic evolution of ligands by exponential enrichment (SELEX) method were discovered by Larry Gold and Andrew Ellington in 1990, respectively. Based on the various advantages of aptamers, they have become a potent rival of antibodies in therapeutics and bio-analysis. Especially, the recent advances in aptamer biosensor application are remarkable due to its intrinsic properties of aptamers as nucleic acids and target induced conformational changes, in addition to the introduction of graphene oxide-based easy and simple immobilization-free screening method even for dual aptamers. In addition, the incorporation of various nanomaterials such as metallic nanoparticles, carbon materials, and functional nanospheres in aptasensors has facilitated the improvement of analytical performance and commercial application of aptasensors. In this review, recent prominent reports on aptasensors utilizing nanomaterials were introduced to understand the principle of aptamer-based biosensors and provide an insight for new strategies of aptasensors and the application of various nanomaterials. The perspective on aptamer-based biosensors and diagnostics was also discussed in view of technology and market.


Subject(s)
Biosensing Techniques/methods , Nanostructures/chemistry , Nanotechnology/methods , SELEX Aptamer Technique/methods , Antibodies/chemistry , Graphite/chemistry , Ligands , Quartz Crystal Microbalance Techniques/methods
4.
ACS Appl Mater Interfaces ; 7(11): 6082-92, 2015 Mar 25.
Article in English | MEDLINE | ID: mdl-25723711

ABSTRACT

In this manuscript, natural materials were combined into a single "pot" to produce flexible, highly fire resistant, and bioinspired coatings on flexible polyurethane foam (PUF). In one step, PUF was coated with a fire protective layer constructed of a polysaccharide binder (starch or agar), a boron fire retardant (boric acid or derivative), and a dirt char former (montmorillonite clay). Nearly all coatings produced a 63% reduction in a critical flammability value, the peak heat release rate (PHRR). One formulation produced a 75% reduction in PHRR. This technology was validated in full-scale furniture fire tests, where a 75% reduction in PHRR was measured. At these PHRR values, this technology could reduce the fire threat of furniture from significant fire damage in and beyond the room of fire origin to being contained to the burning furniture. This flammability reduction was caused by three mechanisms-the gas-phase and condensed-phase processes of the boron fire retardant and the condensed-phase process of the clay. We describe the one-pot coating process and the impact of the coating composition on flammability.


Subject(s)
Biomimetic Materials/chemical synthesis , Boric Acids/chemistry , Fires/prevention & control , Flame Retardants/chemical synthesis , Polyurethanes/chemistry , Bentonite/chemistry , Coated Materials, Biocompatible/chemical synthesis , Hot Temperature , Materials Testing , Polysaccharides/chemistry , Thermal Conductivity
5.
ACS Appl Mater Interfaces ; 6(3): 2146-52, 2014 Feb 12.
Article in English | MEDLINE | ID: mdl-24422757

ABSTRACT

Layer-by-layer (LbL) assembly coatings reduce the flammability of textiles and polyurethane foam but require extensive repetitive processing steps to produce the desired coating thickness and nanoparticle fire retardant content that translates into a fire retardant coating. Reported here is a new hybrid bi-layer (BL) approach to fabricate fire retardant coatings on polyurethane foam. Utilizing hydrogen bonding and electrostatic attraction along with the pH adjustment, a fast growing coating with significant fire retardant clay content was achieved. This hybrid BL coating exhibits significant fire performance improvement in both bench scale and real scale tests. Cone calorimetry bench scale tests show a 42% and 71% reduction in peak and average heat release rates, respectively. Real scale furniture mockups constructed using the hybrid LbL coating reduced the peak and average heat release rates by 53% and 63%, respectively. This is the first time that the fire safety in a real scale test has been reported for any LbL technology. This hybrid LbL coating is the fastest approach to develop an effective fire retardant coating for polyurethane foam.

6.
Biosens Bioelectron ; 54: 195-8, 2014 Apr 15.
Article in English | MEDLINE | ID: mdl-24280049

ABSTRACT

Microbial cells have many binding moieties on their surface for binding to their specific bioreceptors. The whole-cell SELEX process enables the isolation of various aptamers that can bind to different components on the cell surface such as proteins, polysaccharides, or flagella with high affinity and specificity. Here, we examine the binding capacity of an aptamer mixture (aptamer cocktail) composed of various combinations of 3 different DNA aptamers isolated from Escherichia coli and compare it with one of the single aptamers using fluorescence-tagged aptamers. The aptamer mixtures showed higher fluorescence signal than did any single aptamer used, which suggests that use of aptamer mixtures can enhance the sensitivity of detection of microbial cells. To further evaluate this effect, the signal enhancement and improvement of sensitivity provided by combinatorial use of aptamers were examined in an electrochemical detection system. With regard to current decreases, the aptamer cocktail immobilized on gold electrodes performed better than a single aptamer immobilized on gold electrodes did. Consequently, the detection limit achieved using the aptamers individually was approximately 18 times that when the 3 aptamers were used in combination. These results support the use of aptamer cocktails for detection of complex targets such as E. coli with enhanced sensitivity.


Subject(s)
Aptamers, Nucleotide/chemistry , Biosensing Techniques/methods , Escherichia coli/isolation & purification , Escherichia coli/chemistry , Escherichia coli Infections/diagnosis , Humans , SELEX Aptamer Technique/methods , Sensitivity and Specificity
7.
Adv Biochem Eng Biotechnol ; 140: 29-67, 2014.
Article in English | MEDLINE | ID: mdl-23851587

ABSTRACT

It has been 20 years since aptamer and SELEX (systematic evolution of ligands by exponential enrichment) were described independently by Andrew Ellington and Larry Gold. Based on the great advantages of aptamers, there have been numerous isolated aptamers for various targets that have actively been applied as therapeutic and analytical tools. Over 2,000 papers related to aptamers or SELEX have been published, attesting to their wide usefulness and the applicability of aptamers. SELEX methods have been modified or re-created over the years to enable aptamer isolation with higher affinity and selectivity in more labor- and time-efficient manners, including automation. Initially, most of the studies about aptamers have focused on the protein targets, which have physiological functions in the body, and their applications as therapeutic agents or receptors for diagnostics. However, aptamers for small molecules such as organic or inorganic compounds, drugs, antibiotics, or metabolites have not been studied sufficiently, despite the ever-increasing need for rapid and simple analytical methods for various chemical targets in the fields of medical diagnostics, environmental monitoring, food safety, and national defense against targets including chemical warfare. This review focuses on not only recent advances in aptamer screening methods but also its analytical application for small molecules.


Subject(s)
Aptamers, Nucleotide/analysis , Aptamers, Nucleotide/chemistry , Biosensing Techniques/methods , Conductometry/methods , Immunoassay/methods , SELEX Aptamer Technique/methods , Spectrometry, Fluorescence/methods , Aptamers, Nucleotide/genetics , High-Throughput Screening Assays/methods , Small Molecule Libraries/chemical synthesis
8.
Anal Biochem ; 436(1): 22-8, 2013 May 01.
Article in English | MEDLINE | ID: mdl-23357235

ABSTRACT

Aptamers are powerful capturing probes against various targets such as proteins, small organic compounds, metal ions, and even cells. In this study, we isolated and characterized single-stranded DNA (ssDNA) aptamers against Escherichia coli. A total of 28 ssDNAs were isolated after 10 rounds of selection using a bacterial cell-SELEX (systematic evolution of ligands by exponential enrichment) process. Other bacterial species (Klebsiella pneumoniae, Citrobacter freundii, Enterobacter aerogenes, and Staphylococcus epidermidis) were used for counter selection to enhance the selectivity of ssDNA aptamers against E. coli. Finally, four ssDNA aptamers showed high affinity and selectivity to E. coli, The dissociation constants (K(d)) of these four ssDNA aptamers to E. coli were estimated to range from 12.4 to 25.2 nM. These aptamers did not bind to other bacterial species, including four counter cells, but they showed affinity to different E. coli strains. The binding of these four aptamers to E. coli was observed directly by fluorescence microscopy.


Subject(s)
Anti-Bacterial Agents/isolation & purification , Anti-Bacterial Agents/pharmacology , Aptamers, Nucleotide/isolation & purification , Aptamers, Nucleotide/pharmacology , DNA, Single-Stranded/pharmacology , Escherichia coli/drug effects , Anti-Bacterial Agents/chemistry , Aptamers, Nucleotide/chemistry , DNA, Single-Stranded/chemistry , DNA, Single-Stranded/isolation & purification , Escherichia coli/cytology , Ligands , Microbial Sensitivity Tests , Microscopy, Fluorescence , SELEX Aptamer Technique
9.
Appl Biochem Biotechnol ; 168(5): 1143-52, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22948603

ABSTRACT

The bactericidal activity of TiO(2) nanoparticles under visible light is very important in regards to its practical applications. In this paper, we synthesized vanadium-pentoxide-loaded TiO(2) nanoparticles (V(2)O(5)-TiO(2)) using a chemical vapor condensation method, followed by the impregnation method, and characterized its physicochemical properties through X-ray diffraction patterning, X-ray photoelectron spectroscopy analysis, Raman spectra analysis, and Fourier transform infrared analysis. In addition, the antibacterial activity of V(2)O(5)-TiO(2) nanoparticles against E. coli was evaluated and compared with pure TiO(2) nanoparticles. In these experiments, the population of E. coli was shown to be significantly reduced by V(2)O(5)-TiO(2) nanoparticles under illumination with fluorescent light, whereas pure TiO(2) nanoparticles showed about 3.3-fold lower antibacterial activity than the V(2)O(5)-TiO(2) nanoparticles. This result was most likely due to the change in surface conditions of the TiO(2) nanoparticles, which was due to the loading of vanadium pentoxide on the TiO(2) nanoparticles. Furthermore, both photocatalysts showed similar antibacterial activity under UV-A (352 nm) irradiation.


Subject(s)
Anti-Infective Agents , Escherichia coli/drug effects , Nanoparticles , Titanium , Vanadium Compounds , Anti-Infective Agents/chemical synthesis , Anti-Infective Agents/chemistry , Anti-Infective Agents/pharmacology , Anti-Infective Agents/radiation effects , Light , Nanoparticles/chemistry , Nanoparticles/microbiology , Nanoparticles/radiation effects , Photoelectron Spectroscopy , Spectroscopy, Fourier Transform Infrared , Spectrum Analysis, Raman , Titanium/chemistry , Titanium/pharmacology , Vanadium Compounds/chemistry , Vanadium Compounds/pharmacology , X-Ray Diffraction
10.
ACS Macro Lett ; 1(7): 820-824, 2012 Jul 17.
Article in English | MEDLINE | ID: mdl-35607125

ABSTRACT

An innovative twist to fabricating layer-by-layer coatings resulted in transparent, high-content clay coatings on porous polyurethane foam. The addition of an anionic poly(acrylic acid) (PAA) monolayer between anionic clay and cationic branched-polyethylenimine (PEI) monolayers resulted in a trilayer nanocomposite structure with an order of magnitude thicker coating using 40% less monolayers than the conventional bilayer approach. The eight trilayer system thoroughly coated all internal and external surfaces of the porous polyurethane foam, creating a clay brick wall barrier that reduced the foam flammability by as much as 17% of the peak heat release rate and 21% of the total burn time. Though the flammability reduction is comparable to common commercial fire retardant polyurethane foam, the clay is used at a 50% lower amount and may be a greener solution as many of the commercial fire retardants (e.g., halogen bases) have potential environmental and health concerns.

11.
Analyst ; 136(18): 3720-4, 2011 Sep 21.
Article in English | MEDLINE | ID: mdl-21799952

ABSTRACT

We developed a homogeneous fluorescence assay for multiplex detection based on the target induced conformational change of DNA aptamers. DNA aptamers were immobilized on quantum dots (QDs), and QDs conjugated ssDNA was adsorbed on the surface of gold nanoparticles (AuNPs) by electrostatic interaction between uncoiled ssDNA and the AuNPs. Subsequently the fluorescence of QDs was effectively quenched by the AuNPs due to fluorescence resonance energy transfer (FRET) of QDs to AuNPs. In the presence of targets, the QDs conjugated aptamers were detached from AuNPs by target induced conformational change of aptamers, consequently the fluorescence of the QDs was recovered proportional to the target concentration. In this study, three different QD/aptamer conjugates were used for multiplex detection of mercury ions, adenosine and potassium ions. In a control experiment, all of the three targets were simultaneously detected with high selectivity.


Subject(s)
Aptamers, Nucleotide/chemistry , Fluorescence Resonance Energy Transfer , Gold/chemistry , Metal Nanoparticles/chemistry , Adenosine/analysis , DNA, Single-Stranded/chemistry , Ions/chemistry , Mercury/analysis , Potassium/analysis , Quantum Dots , Static Electricity
12.
Macromol Rapid Commun ; 32(15): 1169-73, 2011 Aug 03.
Article in English | MEDLINE | ID: mdl-21748816

ABSTRACT

Small, organic, toxic compounds are not well eliminated by water-treatment systems and eventually become concentrated in the human body. In this study, liposomes are employed to house aptamers with their own binding buffer. When small, organic, toxic compounds in water pass through a liposome barrier, only the target molecules are captured by the DNA aptamers inside the liposomes. The capture efficiency is not high when DNA aptamers are used in tap water. When DNA aptamers in liposomes are used, the capture efficiency increases more than 80%. The simultaneous and selective elimination of target toxicants is successfully performed for tap-water samples containing toxicant mixtures.


Subject(s)
Aptamers, Nucleotide/chemistry , Liposomes/chemistry , Organic Chemicals/chemistry , Water Pollutants, Chemical/chemistry , Water Purification/methods , Adsorption , Water Purification/instrumentation
13.
Biosens Bioelectron ; 26(10): 4058-63, 2011 Jun 15.
Article in English | MEDLINE | ID: mdl-21514814

ABSTRACT

We present an important role of the ratio of affinities in unmodified gold nanoparticles-based colorimetric aptasensor reactions. An affinity ratio, representing the competitive interactions among aptamers, targets, and unmodified gold nanoparticles (umAuNPs), was found to be an important factor for the sensitivity (the performance), where the affinity ratio is the affinity of the aptamer to targets divided by the affinity to umAuNPs (K(dAuNP)/K(dTarget)). In this study, the five different aptamers having different affinity ratios to both umAuNPs and targets are used, and the degree of color change is well correlated with its affinity ratio. This result is verified by using a tetracycline binding aptamer (TBA) showing different affinities to its three derivatives, tetracycline, oxytetracycline and doxycycline. Based on this model, the sensitivity of umAuNPs based colorimetric detection for ibuprofen can be enhanced simply through reducing the ibuprofen binding aptamer's affinity to umAuNP by using bis (p-sulfonatophenyl) phenylphosphine as an AuNP-capping ligand, instead of using the citrate. As a result, a clear color change is observed even at a 20-fold less amount of ibuprofen. This study presents that the performance (detection sensitivity) of umAuNPs-based colorimetric aptasensors could be improved by simply adjusting the affinity ratio of the aptamers to targets and umAuNPs, without knowing the conformational changes of aptamers upon the target binding or needing any modification of aptamer sequences.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques/methods , Binding, Competitive , Colorimetry/methods , Doxycycline/analysis , Gold , Metal Nanoparticles , Oxytetracycline/analysis , Tetracycline/analysis
14.
Appl Biochem Biotechnol ; 164(8): 1278-91, 2011 Aug.
Article in English | MEDLINE | ID: mdl-21409410

ABSTRACT

Silver nanoparticles (AgNPs), exhibiting a broad size range and morphologies with highly reactive facets, which are widely applicable in real-life but not fully verified for biosafety and ecotoxicity, were subjected to report transcriptome profile in yeast Saccharomyces cerevisiae. A large number of genes accounted for ∼3% and ∼5% of the genome affected by AgNPs and Ag-ions, respectively. Principal component and cluster analysis suggest that the different physical forms of Ag were the major cause in differential expression profile. Among 90 genes affected by both AgNPs and Ag-ions, metalloprotein mediating high resistance to copper (CUP1-1 and CUP1-2) were strongly induced by AgNPs (∼45-folds) and Ag-ions (∼22-folds), respectively. A total of 17 genes, responsive to chemical stimuli, stress, and transport processes, were differentially induced by AgNPs. The differential expression was also seen with Ag-ions that affected 73 up- and 161 down-regulating genes, and most of these were involved in ion transport and homeostasis. This study provides new information on the knowledge for impact of nanoparticles on living microorganisms that can be extended to other nanoparticles.


Subject(s)
Metal Nanoparticles/chemistry , Saccharomyces cerevisiae/drug effects , Saccharomyces cerevisiae/genetics , Silver/pharmacology , Cations , Cluster Analysis , Gene Expression Profiling , Gene Expression Regulation, Fungal/drug effects , Metallothionein/genetics , Mutagenicity Tests , Principal Component Analysis , Saccharomyces cerevisiae Proteins/genetics
15.
Biosens Bioelectron ; 26(4): 1644-9, 2010 Dec 15.
Article in English | MEDLINE | ID: mdl-20829027

ABSTRACT

We have successfully developed a novel colorimetric aptasensor using gold nanoparticles for highly sensitive and specific detection of oxytetracycline (OTC), one of the most common antibacterial agents. A highly specific ssDNA aptamer that bind to OTC with high affinity was employed to discriminate other tetracyclines (TCs), such as doxycycline (DOX) and tetracycline (TET). Aggregation of AuNPs was specifically induced by desorption of the OTC binding aptamers (OBAs) from the surface of gold nanoparticles as a result of the aptamer-target interaction, leading to the color change from red to purple. The detection limit of OTC was enhanced up to 25 nM, which is 20-fold lower than the limit USA-EPA regulated, with two orders of magnitudes in its linear dynamic range by successful optimization on the amount of the aptamers, AuNPs, and salts. This colorimetric aptasensor is advantageous over the other conventional methods in terms of its simple signal generation and detection with the naked eye, which can be realized in on-site detection of antibacterial agents.


Subject(s)
Anti-Bacterial Agents/analysis , Aptamers, Nucleotide , Biosensing Techniques/methods , Colorimetry/methods , Metal Nanoparticles , Oxytetracycline/analysis , Animals , Anti-Bacterial Agents/chemistry , Aptamers, Nucleotide/chemistry , Aptamers, Nucleotide/genetics , Base Sequence , Biosensing Techniques/statistics & numerical data , Food Contamination/analysis , Gold , Humans , Oxytetracycline/chemistry , Sensitivity and Specificity
16.
Bioorg Med Chem ; 18(10): 3467-73, 2010 May 15.
Article in English | MEDLINE | ID: mdl-20456958

ABSTRACT

Single stranded DNA aptamers that can bind to ibuprofen, a widely used anti-inflammation drug, were selected from random DNA library of 10(15) nucleotides by FluMag-SELEX process. Five different sequences were selected and their enantioselectivity and affinity were characterized. Three out of five aptamer candidates did not show any affinity to (S)-ibuprofen, but only to racemic form of ibuprofen, suggesting that they are (R)-ibuprofen specific aptamers. Another two aptamer candidates showed affinity to both racemic form and (S)-ibuprofen, which were considered as (S)-ibuprofen specific aptamers. The affinity of five ssDNA aptamers isolated was in a range of 1.5-5.2microM. In addition, all of these five aptamers did not show any affinity to analogues of ibuprofen in its profen's group (fenoprofen, flubiprofen, and naproxen) and the antibiotics of oxytetracycline, another control.


Subject(s)
Aptamers, Nucleotide/isolation & purification , DNA, Single-Stranded/metabolism , Ibuprofen/chemistry , Nucleic Acid Conformation/drug effects , Oligonucleotides/chemistry , Aptamers, Nucleotide/chemistry , Base Sequence/drug effects , Binding Sites , DNA, Bacterial , Gene Library , Ibuprofen/pharmacology , Kinetics , Molecular Sequence Data , SELEX Aptamer Technique , Structure-Activity Relationship
17.
Bioprocess Biosyst Eng ; 33(1): 31-7, 2010 Jan.
Article in English | MEDLINE | ID: mdl-19701778

ABSTRACT

An electrochemical aptasensor was developed for the detection of tetracycline using ssDNA aptamer that selectively binds to tetracycline as recognition element. The aptamer was highly selective for tetracycline which distinguishes minor structural changes on other tetracycline derivatives. The biotinylated ssDNA aptamer was immobilized on a streptavidin-modified screen-printed gold electrode, and the binding of tetracycline to aptamer was analyzed by cyclic voltammetry and square wave voltammetry. Our results showed that the minimum detection limit of this sensor was 10 nM to micromolar range. The aptasensor showed high selectivity for tetracycline over the other structurally related tetracycline derivatives (oxytetracycline and doxycycline) in a mixture. The aptasensor developed in this study can potentially be used for detection of tetracycline in pharmaceutical preparations, contaminated food products, and drinking water.


Subject(s)
Aptamers, Nucleotide/chemistry , DNA, Single-Stranded/chemistry , Electrochemical Techniques/methods , Tetracycline/analysis , Electrodes , Food Analysis/methods , Gold/chemistry , Sensitivity and Specificity , Streptavidin/chemistry
18.
Anal Chim Acta ; 634(2): 250-4, 2009 Feb 23.
Article in English | MEDLINE | ID: mdl-19185128

ABSTRACT

An electrochemical sensing system for oxytetracycline (OTC) detection was developed using ssDNA aptamer immobilized on gold interdigitated array (IDA) electrode chip. A highly specific ssDNA aptamer that bind to OTC with high affinity was employed to discriminate other tetracyclines (TCs), such as doxycycline (DOX) and tetracycline (TET). The immobilized thiol-modified aptamer on gold electrode chip served as a biorecognition element for the target molecules and the electrochemical signals generated from interactions between the aptamers and the target molecules was evaluated by cyclic voltammetry (CV) and square wave voltammetry (SWV). The current decrease due to the interference of bound OTC, DOX or TET was analyzed with the electron flow produced by a redox reaction between ferro- and ferricyanide. The specificity of developed EC-biosensor for OTC was highly distinguishable from the structurally similar antibiotics (DOX and TET). The dynamic range was determined to be 1-100 nM of OTC concentration in semi-logarithmic coordinates.


Subject(s)
Aptamers, Nucleotide/analysis , Aptamers, Nucleotide/chemistry , Microarray Analysis/instrumentation , Microarray Analysis/methods , Oxytetracycline/analysis , Electrochemistry , Electrodes , Gold/chemistry , Molecular Structure , Reproducibility of Results
20.
Anal Chem ; 80(8): 2867-73, 2008 Apr 15.
Article in English | MEDLINE | ID: mdl-18324839

ABSTRACT

Retinol binding protein 4 (RBP4) is a useful biomarker in the diagnosis of type 2 diabetes since its level in the serum is higher in insulin-resistant states. Accurate measurement of the serum RBP4 levels is hampered by conventional immunologic methods, such as enzyme-linked immunosorbent assay (ELISA). In this study, therefore, we have developed an aptamer-based surface plasmon resonance (SPR) biosensor that can be used to sense for RBP4 in serum samples. A single-stranded DNA (ssDNA) aptamer that showed high affinity (Kd = 0.2 +/- 0.03 microM) and specificity to RBP4 was selected. This RBP4-specific aptamer was immobilized on a gold chip and used in a label-free RBP4 detection using SPR. Analysis of RBP4 in artificial serum using SPR was compared with ELISA and Western blot analysis. Our results indicated that the RBP4-specific aptamer-based SPR biosensor gave better dose-dependent responses and was more sensitive than ELISA assays. As such, this RBP4 aptamer-based SPR biosensor can be potentially used to monitor the RBP4 levels within the serum as an indicator of type 2 diabetes.


Subject(s)
Aptamers, Nucleotide/chemistry , DNA, Single-Stranded/chemistry , Diabetes Mellitus, Type 2/blood , Retinol-Binding Proteins, Plasma/analysis , Surface Plasmon Resonance/methods , Animals , Aptamers, Nucleotide/genetics , Aptamers, Nucleotide/metabolism , Base Sequence , Biosensing Techniques/methods , Blotting, Western , Cattle , DNA, Single-Stranded/genetics , DNA, Single-Stranded/metabolism , Diabetes Mellitus, Type 2/diagnosis , Enzyme-Linked Immunosorbent Assay , Humans , Kinetics , Molecular Sequence Data , Nucleic Acid Conformation , Retinol-Binding Proteins, Plasma/genetics , Retinol-Binding Proteins, Plasma/metabolism , Substrate Specificity
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