Adsorption Of Aptamer Research Articles

Ultrathin metal-organic framework nanosheets (Cu-TCPP) assembled micro flower combined with endonuclease signal amplification for ultrasensitive detection

Metal-organic framework play an important role in developing novel detection techniques. Herein, an ultrathin two-dimensional (2D) metal-organic framework nanosheet (Cu-TCPP) assembled microflower was successfully synthesized and applied for ultrasensitive detection. The high detection performance of Cu-TCPP microflower was attributed to high-capacity aptamer adsorption and specific desorption resulting from numerous approachable active sites, and its ultrathin nanosheets microflower-like structure. Moreover, the Cu-TCPP microflower had overcame the disadvantages of 2D MOF nanosheets which tended to agglomerate. Moreover, it exhibited excellent fluorescence quenching performance as a donor for fluorescence resonance energy transfer. Acetamiprid, a new chloronicotinic neurotoxic insecticide was used as the model analyte. The ultrasensitive sensor was developed by combining the Cu-TCPP microflower with deoxyribonuclease Ⅰ signal enhanced fluorescence. When acetamiprid was present, the fluorescent labeled aptamer probe was selectively bound to acetamiprid and desorbed from the Cu-TCPP microflower, resulting in fluorescence recovery. Subsequently, DNase Ⅰ was used to cleave the DNA strands of the aptamer probe, and acetamiprid was released to participate in a new cycle. The detection limit of the assay was as low as 5.56 pg/mL with a linear range 50.00 pg/mL∼5.00 μg/mL, and it was a universal platform to detect other analytes.

Mg2+- or Ca2+-regulated aptamer adsorption on polydopamine-coated magnetic nanoparticles for fluorescence detection of ochratoxin A.

It has been observed that polyvalent metal ions can mediate the adsorption of DNA on polydopamine (PDA) surfaces. Exploiting this, we used two divalent metal ions (Mg2+ or Ca2+) to promote the adsorption of fluorescence-labelled ochratoxin A (OTA) aptamers on PDA-coated magnetic nanoparticles (Fe3O4@PDA). Based on the different adsorption affinities of free aptamers and OTA-bound aptamers, a facile assay method was established for OTA detection. The aptamers adsorbed on Fe3O4@PDA were removed via simple magnetic separation, and the remaining aptamers in the supernatant exhibited a positive correlation with the OTA concentration. The concentrations of Mg2+ and Ca2+ were finely tuned to attain the optimal adsorption affinity and sensitivity for OTA detection. In addition, other factors, including the Fe3O4@PDA dosage, pH, mixing order, and incubation time, were studied. Finally, under optimized conditions, a detection limit (3σ/s) of 1.26ng/mL was achieved for OTA. Real samples of spiked red wine were analysed with this aptamer-based method. This is the first report of regulating aptamer adsorption on the PDA surface with polyvalent metal ions for OTA detection. By changing the aptamers, the method can be easily extended to other target analytes.

Adsorption of tetracycline antibiotics to gold nanoparticles and feasibility of aptamer-based label-free colorimetric detection

Tetracyclines are a group of very important antibiotics that are still in use. To extract, detect, and remove tetracyclines from the environment, various nanomaterials have been employed. Although gold nanoparticles (AuNPs) are a commonly cited material for these purposes, a fundamental understanding of these tetracycline-AuNP systems is still limited. In this work, the adsorption of tetracycline, oxytetracycline, and doxycycline to AuNPs was studied. The effect on the colloidal stability of AuNPs, adsorption kinetics, and the resulting adsorption isotherms was measured. While millimolar concentrations of the tetracyclines can cause aggregation of AuNPs, saturated monolayer adsorption was achieved with low micromolar concentrations of the tetracyclines. Adsorption was instantaneous, and adsorption to AuNPs enhanced their intrinsic fluorescence instead of quenching. With the assumption that aptamer/target complexes cannot be easily adsorbed by AuNPs compared to free aptamers, a label-free colorimetric detection method was tested. While the label-free sensor showed target-dependent aggregation of AuNPs, a nonbinding mutant aptamer showed the same trend, suggesting that the color change did not reflect aptamer adsorption but other events such as target adsorption. This study indicates the importance of the fundamental understanding of target/AuNP interactions to correctly design aptamer and AuNP-based label-free biosensors.

Target-induced gold nanoparticles colorimetric sensing coupled with aptamer for rapid and high-sensitivity detecting kanamycin

Herein we report a novel colorimetric sensing strategy for the detection of kanamycin (kana) based on target-induced gold nanoparticles (AuNPs) coupled with aptamers. Aptamer-functionalized AuNPs, as the colorimetric probe, showed a distinct red shift with addition of kana, which avoiding the tedious and unnecessary additive-induced process. To study the interaction between kana and AuNPs and the effects of the specific aptamer adsorption, a series of experiments including UV–vis absorbance and surface enhanced Raman spectroscopy (SERS) were performed. Based on the results, a new alternative view is proposed that kana can directly induce the aggregation of aptamer-wrapped AuNPs, attributed to the co-adsorption of kana and aptamer on the surface of AuNPs. The proposed colorimetric sensing exhibited high selectivity and sensitivity for kanamycin assay with a wide linear range from 10.0 nM to 4.0 μM, and the limit of detection (LOD) reached 4.0 nM. Moreover, the whole detection process could be completed within 5 min, and it also achieved excellent performance in real samples detection with recoveries in the range of 86.22–109.89%. The results indicate that target-induced AuNPs colorimetric sensing coupled with aptamers for the direct detection of kana is simple, rapid and high-sensitivity, has the promising potential applications in the fields of food safety and environmental monitoring.

Reversible and Irreversible HAuCl4 Binding to DNA for Seeded Gold Nanoparticle Growth and Opposite DNA and Aptamers Colorimetric Sensing Outcomes

AbstractDNA‐directed seeded growth of gold nanoparticles has been used for the development of aptamer‐based biosensors with the assumption that target analytes can modulate the adsorption of aptamers to the gold seeds and thus affect the growth reaction. To understand the reaction, the effect of single‐ and double‐stranded DNA is first examined, and it is found that they have a similar promotion effect of anisotropic growth, suggesting that DNA cannot be detected using this method. By studying the interaction between HAuCl4 and DNA, both weak reversible binding and strong irreversible binding are identified, with the latter becoming dominating with a longer incubation time. Single‐ and double‐stranded DNA have similar weak binding to HAuCl4, and this weak binding is more important for the growth reaction. Then three aptamers are tested, where only cortisol appeared to modulate its aptamer adsorption and the growth reaction reflected aptamer binding. Hg2+ shows no advantage for its aptamer, and quinine induced aggregation of AuNPs cannot be detected by this reaction either. Therefore, each aptamer target needs to be individually studied to test if this method is applicable. It is also noted that DNA and aptamers have opposite outcomes for the target‐dependent growth reactions.

A FRET-based aptasensor for the detection of α-synuclein oligomers as biomarkers of Parkinson's disease.

Soluble oligomers of α-synuclein (α-syn) are known to be responsible for neuronal death in the early stages of Parkinson's disease (PD). Thus, the development of a simple, rapid, and inexpensive method for the detection of α-syn oligomers can help PD diagnosis before irreversible damage to the brain tissue occurs. The present study is aimed at developing a FRET-based aptasensor for the selective and sensitive detection of α-syn oligomers. Herein, FAM-labeled aptamer adsorption on graphene oxide (GO) resulted in fluorescence quenching of FAM. The binding of α-syn oligomers to the aptamer led to the recovery of the fluorescence intensity. Under optimized conditions, the developed biosensor showed two linear response ranges from 10-100 nM and 250 nM to 2 μM in α-syn oligomer detection. The limit of detection (LOD) and limit of quantification (LOQ) were calculated to be 6.3 nM and 19.3 nM, respectively. Furthermore, the performance of the sensing platform in the detection of the target analyte in biological matrices was demonstrated by the assay of α-syn oligomers in spiked human saliva samples. According to the obtained results, this sensing platform has a good performance for α-syn oligomer detection and it can be considered as a promising candidate for the early diagnosis of PD.

Lipid Specific Membrane Interaction of Aptamers and Cytotoxicity.

We aim to discover diagnostic tools to detect phosphatidylserine (PS) externalization on apoptotic cell surface using PS binding aptamers, AAAGAC and TAAAGA, and hence to understand chemotherapy drug efficacy when inducing apoptosis into cancer cells. The entropic fragment-based approach designed aptamers have been investigated to inspect three aspects: lipid specificity in aptamers’ membrane binding and bilayer physical properties-induced regulation of binding mechanisms, the apoptosis-induced cancer cell surface binding of aptamers, and the aptamer-induced cytotoxicity. The liposome binding assays show preferred membrane binding of aptamers due to presence of PS in predominantly phosphatidylcholine-contained liposomes. Two membrane stiffness reducing amphiphiles triton X-100 and capsaicin were found to enhance membrane’s aptamer adsorption suggesting that bilayer physical properties influence membrane’s adsorption of drugs. Microscopic images of fluorescence-tagged aptamer treated LoVo cells show strong fluorescence intensity only if apoptosis is induced. Aptamers find enhanced PS molecules to bind with on the surface of apoptotic over nonapoptotic cells. In cytotoxicity experiments, TAAAGA (over poor PS binding aptamer CAGAAAAAAAC) was found cytotoxic towards RBL cells due to perhaps binding with nonapoptotic externalized PS randomly and thus slowly breaching plasma membrane integrity. In these three experimental investigations, we found aptamers to act on membranes at comparable concentrations and specifically with PS binding manner. Earlier, we reported the origins of actions through molecular mechanism studies—aptamers interact with lipids using mainly charge-based interactions. Lipids and aptamers hold distinguishable charge properties, and hence, lipid–aptamer association follows distinguishable energetics due to electrostatic and van der Waals interactions. We discover that our PS binding aptamers, due to lipid-specific interactions, appear as diagnostic tools capable of detecting drug-induced apoptosis in cancer cells.

Changes of Viscoelastic Properties of Aptamer-Based Sensing Layers Following Interaction with Listeria innocua.

A multiharmonic quartz crystal microbalance (QCM) has been applied to study the viscoelastic properties of the aptamer-based sensing layers at the surface of a QCM transducer covered by neutravidin following interaction with bacteria Listeria innocua. Addition of bacteria in the concentration range 5 × 103–106 CFU/mL resulted in a decrease of resonant frequency and in an increase of dissipation. The frequency decrease has been lower than one would expect considering the dimension of the bacteria. This can be caused by lower penetration depth of the acoustics wave (approximately 120 nm) in comparison with the thickness of the bacterial layer (approximately 500 nm). Addition of E. coli at the surface of neutravidin as well as aptamer layers did not result in significant changes in frequency and dissipation. Using the Kelvin–Voight model the analysis of the viscoelastic properties of the sensing layers was performed and several parameters such as penetration depth, Γ, viscosity coefficient, η, and shear modulus, μ, were determined following various modifications of QCM transducer. The penetration depth decreased following adsorption of the neutravidin layer, which is evidence of the formation of a rigid protein structure. This value did not change significantly following adsorption of aptamers and Listeria innocua. Viscosity coefficient was higher for the neutravidin layer in comparison with the naked QCM transducer in a buffer. However, a further increase of viscosity coefficient took place following attachment of aptamers suggesting their softer structure. The interaction of Listeria innocua with the aptamer layer resulted in slight decrease of viscosity coefficient. The shearing modulus increased for the neutravidin layer and decreased following aptamer adsorption, while a slight increase of µ was observed after the addition of Listeria innocua.

Nanozyme Sensor Array Plus Solvent-Mediated Signal Amplification Strategy for Ultrasensitive Ratiometric Fluorescence Detection of Exosomal Proteins and Cancer Identification.

Tumor exosomes with molecular marker-proteins inherited from their parent cells have emerged as a promising liquid biopsy biomarker for cancer diagnosis. However, facile, robust, and sensitive detection of exosomal proteins remains challenging. Therefore, a nanozyme sensor array is constructed by using aptamer-modified C3N4 nanosheets (Apt/C3N4 NSs) together with a solvent-mediated signal amplification strategy for ratiometric fluorescence detection of exosomal proteins. Three aptamers specific to exosomal proteins are selected to construct Apt/C3N4 NSs for high specific recognition of exosomal proteins. The adsorption of aptamers enhances the catalytic activity of C3N4 NSs as a nanozyme for oxidation of o-phenylenediamine (oPD) to 2,3-diaminophenazine (DAP). In the presence of target exosomes, the strong affinity between aptamer and exosome leads to the disintegration of Apt/C3N4 NSs, resulting in a decrease of catalytic activity, thereby reducing the production of DAP. The ratiometric fluorescence signal based on a photoinduced electron transfer (PET) effect between DAP and C3N4 NSs is dependent on the concentration of DAP generated, thus achieving highly facile and robust detection of exosomal proteins. Remarkably, the addition of organic solvent-1,4-dioxane can sensitize the luminescence of DAP without affecting the intrinsic fluorescence of C3N4 NSs, achieving the amplification of the aptamer-exosome recognition events. The detection limit for exosome is 2.5 × 103 particles/mL. In addition, the accurate identification of cancer can be achieved by machine learning algorithms to analyze the difference of exosomal proteins from different patients' blood. We hope that this facile, robust, sensitive, and versatile nanozyme sensor array would become a promising tool in the field of cancer diagnosis.

Label‐Free Colorimetric Biosensors Based on Aptamers and Gold Nanoparticles: A Critical Review

AbstractTaking advantage of the adsorption of single‐stranded DNA oligonucleotides by gold nanoparticles (AuNPs) and the protection effect of the adsorbed DNA against salt‐induced aggregation of AuNPs, a label‐free colorimetric sensor for the detection of DNA was reported in 2004. Since then, the range of target molecules has extended from complementary nucleic acids to metal ions and small molecules by using aptamers. In the presence of target molecules, a blue color arising from aggregated AuNPs is expected. However, these sensors only considered aptamer binding to its target, and the adsorption of aptamers by AuNPs, while the target/AuNP interactions were ignored. We recently found that target adsorption can often dominate the system. In this Review, we list literature examples of using this label‐free strategy for sensing aptamer targets. Seven target analytes are discussed in detail. For As(III), dopamine, melamine, kanamycin, adenosine, and ATP, target adsorption dominated, and the same color change was observed even with non‐aptamer sequences. Only in the case of K+ detection, did the effect of specific aptamer binding dominate, attributable to weak K+/AuNP interactions. These examples call for a careful evaluation of target adsorption and the use of non‐aptamer control sequences in validating these sensors.

Multiple adsorption properties of aptamers on metal-organic frameworks for nucleic acid assay

Enrichment and detection of circulating free nucleic acids in biological samples have gained great attention for disease diagnosis or prognostic evaluation. Nanoscale metal-organic frameworks (NMOFs) have been used for aptamer-based nucleic acid sensing. In this work, different NMOFs, including ZIF-8, MIL-88, MIL-100, MIL-101, as well as Eu-TDA and Tb-TDA [prepared by the coordination of 2,2′-thiodiacetic acid (TDA) and Eu3+ or Tb3+], were investigated in nucleic acid sensing by employing their aptamer adsorption ability and fluorescence quenching capacity for the labeled dyes. Two types of dye aptamer, FAM-labeled aptamer (FAM-Ap) and TexasRedaptamer (TexasRed-Ap) were designed, and their adsorption properties on NMOFs-were compared. It was found that the TexasRed-Ap can be well used for nucleic acid (miR-21) extraction and sensing by linking with a pH-responsive nucleotide chain (TexasRed-Ap-pH) or with an additional random chain ssDNA-1′ (TexasRed-Ap-a). After interacted with the target miR-21 in biosamples, the TexasRed-dsDNA + NMOFs composites can be collected, and the formed TexasRed-dsDNA can be released by changing pH value or addition of ssDNA-1, which is matched with ssDNA-1′. A linear relationship from 0.1 to 200 pM for miR-21 detection was obtained. The results show that the NMOFs can be used as promising platforms for nucleic acid extraction and fluorescent sensing.

Kanamycin Adsorption on Gold Nanoparticles Dominates Its Label-Free Colorimetric Sensing with Its Aptamer.

A short kanamycin-binding aptamer has been widely used for detecting kanamycin. One of the popular signaling methods is based on the color change of gold nanoparticles (AuNPs) to develop label-free colorimetric biosensors. The general perception was that aptamer binding to its target would inhibit aptamer adsorption by the AuNPs. This inhibited adsorption results in the aggregation of the AuNPs and a color change upon addition of salt. However, the potential adsorption of kanamycin was ignored. Herein, we carefully studied the adsorption of kanamycin on AuNPs and performed a comprehensive analysis using two mutated aptamers and a randomly sequenced DNA which were not supposed to bind kanamycin. In addition, a total of six antibiotics were studied over a wide concentration range. As low as 90 nM kanamycin can induce the aggregation of 3 nM citrate-capped AuNPs, indicating very strong adsorption of kanamycin. The color change was independent of DNA sequence, and all the tested sequences showed a similar color response, regardless of aptamer. Among the different antibiotics, kanamycin and streptomycin induced a color change but not the other four. Our results support an alternative mechanism that kanamycin and streptomycin adsorption by the AuNPs was the main reason for the color change instead of aptamer binding.

Dopamine and Melamine Binding to Gold Nanoparticles Dominates Their Aptamer-Based Label-Free Colorimetric Sensing.

Target-directed aptamer adsorption by gold nanoparticles (AuNPs) has been widely used to develop label-free colorimetric biosensors. However, the potential interactions between target molecules and AuNPs have not been considered, which may lead to misinterpretation of analytical results. In this work, the detection of dopamine, melamine, and K+ was studied as model systems to address this problem. First, dopamine and two control molecules all induced the aggregation of citrate-capped AuNPs with apparent Kd's of 5.8 μM dopamine, 51.6 μM norepinephrine, and 142 μM tyramine. Isothermal titration calorimetry measured the aptamer Kd to be 1.9 μM dopamine and 16.8 μM norepinephrine, whereas tyramine cannot bind. Surface enhanced Raman spectroscopy confirmed direct adsorption of dopamine, and the adsorbed dopamine inhibited the adsorption of DNA. Using a typical salt-induced colorimetric detection protocol, a similar color response was observed regardless of the sequence of DNA, indicating the observed color change reflected the adsorption of dopamine by the AuNPs instead of the binding of dopamine by the aptamer. For this label-free sensor to work, the interaction between the target molecule and AuNPs should be very weak, while dopamine represents an example of strong interactions. For the other two systems, the melamine detection did not reflect aptamer binding either but the K+ detection did, suggesting melamine also strongly interacted with AuNPs, whereas K+ had very weak interactions with AuNPs. Since each target molecule is different, such target/AuNP interactions need to be studied case-by-case to ensure the sensing mechanism.

A comparison of the performance of voltammetric aptasensors for glycated haemoglobin on different carbon nanomaterials-modified screen printed electrodes

The integration of carbon nanomaterials into electrochemical aptasensors has gained significant interest in the recent years because of their high electrical conductivity, mechanical strength, and large surface area. However, no comparative study has been reported so far between different carbon nanomaterials for aptasensing applications. Here, we report, a comparative investigation of six carbon electrode materials (carbon, graphene (G), graphene oxide (GO), multi-wall carbon nanotube (MWCNT), single walled carbon nanotube (SWCNT) and carbon nanofiber (CNF)) on the performance of glycated haemoglobin (HbA1c) aptasensor prepared by physical adsorption. The aptamers were non-covalently immobilized on the six nanomaterial electrodes via π-π stacking interactions between the DNA nucleobases and the surface of the carbon material which creates a barrier to the electron transfer. However, upon binding of the target protein to the aptamer, the aptamer dissociates from the surface leading to enhancement of the electron transfer which represent the basis of the detection. The aptamer adsorption, sensors responses and selectivity of the different nanomaterials were compared showing better performance of the SWCNT-based sensor. The voltammetric SWCNT aptasensors showed high sensitivity and selectivity with detection limits of 0.13 pg/mL and 0.03 pg/mL for total haemoglobin (tHb) and HbA1c, respectively. The aptasensor showed selectivity against other proteins in the blood including cystic fibrosis transmembrane conductance regulator (CFTR), survival motor neuron (SMN), dedicator of cytokinesis 8 (DOCK8), signal transducer and activator of transcription 3 (STAT3). This SWCNT aptasensor was superior to the reported detection assays for HbA1c in terms of sensitivity, selectivity and cost. Moreover, our results demonstrate that the choice of the carbon nanomaterial can have a profound impact on the biosensing performance.

Molecular Dynamics Investigation of the Interactions Between RNA Aptamer and Graphene-Monoxide/Boron-Nitride Surfaces: Applications to Novel Drug Delivery Systems

The interactions between RNA aptamer and boron nitride/graphene monoxide nanosheets were investigated using the molecular dynamics simulations. The potential capability of graphene monoxide and boron nitride surfaces to immobilize RNA aptamer was examined in detail. The distance between center of mass of RNA aptamer and the considered surfaces and root mean square deviation and fluctuation were calculated. The results suggest that the adsorption of RNA aptamer on the boron nitride surface is easily occurred compared to the adsorption on the graphene monoxide surface. Besides, RNA aptamer adsorption on the graphene monoxide nanosheet is energetically more favorable than that on the boron nitride one. The water molecules dipole moment and density profile were used to analyze water effect on the immobilization of aptamer on the GMO and BN surfaces. The results of all-atom molecular dynamics simulations show the higher ability of BN nanosheet for delivery application of this RNA aptamer.

Colorimetric Detection of MPT64 Antibody Based on an Aptamer Adsorbed Magnetic Nanoparticles for Diagnosis of Tuberculosis.

We have developed a colorimetric biosensing system for the detection of antibody against MPT64, a protein secreted by Mycobacterium tuberculosis, using aptamer DNA adsorbed Fe₃O₄ magnetic nanoparticles (MNPs) for diagnosis of tuberculosis (TB). In this system, MNPs were first incubated with single stranded (ss) DNA-type aptamer having a high affinity toward target antibody against MPT64 (anti-MPT64), resulting in quick inhibition of the peroxidase-like activity of MNPs via the adsorption of aptamer on the surface of MNPs. By the addition of sample solutions containing anti-MPT64, aptamer bound on the surface of MNPs would strongly interact with free anti-MPT64 and be detached from the MNPs, thereby increasing the available surface area of the MNPs and consequently yielding enhanced peroxidase activity. Using this strategy, target anti-MPT64 was successfully detected by displaying increased colorimetric intensities from the higher oxidation of employed peroxidase substrate, 2,2'-azino-bis(3-ethylbenzo-thiazoline-6-sulfonic acid) (ABTS). Based on these results, we anticipate that aptamer adsorbed MNPs can serve as a potent probe system for the detection of clinically important target molecules.

Anti-fibronectin aptamers improve the colonization of chitosan films modified with D-(+) Raffinose by murine osteoblastic cells.

The aim of the present study was to investigate how the enrichment of chitosan films with anti-fibronectin aptamers could enhance scaffold colonization by osteoblasts, by improving their adhesion and accelerating their proliferation. Chitosan discs were enriched with excess of anti-fibronectin aptamer. Aptamer adsorption on chitosan was monitored by measuring aptamer concentration in the supernatant by spectrophotometry, as well as its release, while functionalization was confirmed by labelling aptamers with a DNA intercalating dye. Chitosan samples were then characterized morphologically with atomic force microscopy and physically with contact angle measurement. Chitosan enrichment with fibronectin was then investigated by immunofluorescence and Bradford assay. 2% chitosan discs were then enriched with increasing doses of aptamers and used as culture substrates for MC3T3-E1 cells. Cell growth was monitored by optical microscopy, while cell viability and metabolic activity were assessed by chemiluminescence and by Resazurin Sodium Salt assay. Cell morphology was investigated by cytofluorescence and by scanning electron microscopy. Chitosan films efficiently bound and retained aptamers. Aptamers did not affect the amount of adsorbed fibronectin, but affected osteoblasts behavior. Cell growth was proportional to the amount of aptamer used for the functionalization, as well as aptamers influenced cell morphology and their adhesion to the substrate. Our results demonstrate that the enrichment of chitosan films with aptamers could selectively improve osteoblasts behavior. Furthermore, our results support further investigation of this type of functionalization as a suitable modification to ameliorate the biocompatibility of biomaterial for hard tissue engineering applications.

A fluorometric aptamer based assay for cytochrome C using fluorescent graphitic carbon nitride nanosheets

A method is introduced for the fluorometric turn-on detection of cytochrome C (cyt c) by using a specific aptamer and nanosheets composed of fluorescent graphitic carbon nitride (g-C3N4) nanosheets. Bulk g-C3N4 was obtained by pyrolysis of melamine at 550 °C. The nanosheets were characterized by scanning electron microscopy and X-ray diffraction. The material was then subjected to protonation and exfoliation to obtain blue fluorescent g-C3N4 nano-sheets. The adsorption of aptamers on the surface of the g-C3N4 nanosheets leads to quenching of its fluorescence (measured at excitation/emission wavelengths of 320/450 nm), but fluorescence is restored on addition of cyt c. Response is linear in the 16 to 140 nM cyt c concentration window, and the detection limit is as low as 2.6 nM. The assay is simple, inexpensive, and can be easily performed. It is selective for cyt c over a number of interfering species. The method was successfully applied to the determination of cyt c in spiked human serum samples.

Colorimetric determination of 17β-estradiol based on the specific recognition of aptamer and the salt-induced aggregation of gold nanoparticles

An aptamer-based colorimetric method for the detection of 17β-estradiol was established by employing AuNPs as the colorimetric probe. This assay was based on the principle that the salt-induced aggregation of AuNPs could be controlled by the adsorption of aptamers on the surface of AuNPs, which was modulated by the specific recognition of aptamers with targets. The method was successfully applied for tap water samples with satisfactory recoveries, which were in full consistence with the results from HPLC. The response of 17β-estradiol was much higher than that of other endocrine-disrupting compounds and several probably coexisting chemicals. The advantages of simplicity, rapidity and selectivity, make this method a useful tool for the determination of 17β-estradiol in real samples.

Highly sensitive colorimetric detection of 17β-estradiol using split DNA aptamers immobilized on unmodified gold nanoparticles.

Gold nanoparticle (AuNP) based colorimetric aptasensor have been developed for many analytes recently largely because of the ease of detection, high sensitivity, and potential for high-throughput analysis. Most of the target aptamers for detection have short sequences. However, the approach shows poor performance in terms of detection sensitivity for most of the long-sequence aptamers. To address this problem, for the first time, we split the 76 mer aptamer of 17β-estradiol into two short pieces to improve the AuNP based colorimetric sensitivity. Our results showed that the split P1 + P2 still retained the original 76 mer aptamer's affinity and specificity but increased the detection limit by 10-fold, demonstrating that as low as 0.1 ng/mL 17β-estradiol could be detected. The increased sensitivity may be caused by lower aptamer adsorption concentration and a lower affinity to the AuNPs of a short single-strand DNA (ssDNA) sequence. Our study provided a new way to use long-sequence aptamers to develop a highly sensitive AuNP-based colorimetric aptasensor.