Electrochemical Detection of Cortisol by Silver Nanoparticle-Modified Molecularly Imprinted Polymer-Coated Pencil Graphite Electrodes.

The sensitive cortisol detection by an electrochemical sensor based on silver nanoparticle-doped molecularly imprinted polymer was successfully improved. This study describes the method development for cortisol detection in both aqueous solution and biological samples using molecularly imprinted poly(hydroxyethyl methacrylate-N-methacryloyl-(l)-histidine methyl ester)-coated pencil graphite electrodes modified with silver nanoparticles (AgNPs) by differential pulse voltammetry (DPV). The cortisol-imprinted pencil graphite electrode (PGE) has a large surface area because of doped AgNPs with enhanced electroactivity. The prepared molecularly imprinted polymer was characterized by scanning electron microscopy. The DPV response of the synthesized electrode with outstanding electrical conductivity was clarified. Cortisol-imprinted polymer-coated PGEs (MIP), cortisol-imprinted polymer-coated PGEs with AgNPs (MIP@AgNPs), and nonimprinted polymer-coated PGEs with AgNPs (NIP@AgNPs) were evaluated for sensitive and selective detection of cortisol in aqueous solution. Five different cortisol concentrations (0.395, 0.791, 1.32, 2.64, and 3.96 nM) were applied to the MIP@AgNPs, and signal responses were detected by the DPV with a regression coefficient (R2) value of 0.9951. The modified electrode showed good electrocatalytic activity toward cortisol for the linear concentration range from 0.395 to 3.96 nM, and a low limit of detection was recorded as 0.214 nM. The results indicate that the MIP@AgNPs sensor has great potential for sensitive and selective cortisol determination in biological samples.

Aptamer-Based Point-of-Care Devices: Emerging Technologies and Integration of Computational Methods.

Recent innovations in point-of-care (POC) diagnostic technologies have paved a critical road for the improved application of biomedicine through the deployment of accurate and affordable programs into resource-scarce settings. The utilization of antibodies as a bio-recognition element in POC devices is currently limited due to obstacles associated with cost and production, impeding its widespread adoption. One promising alternative, on the other hand, is aptamer integration, i.e., short sequences of single-stranded DNA and RNA structures. The advantageous properties of these molecules are as follows: small molecular size, amenability to chemical modification, low- or nonimmunogenic characteristics, and their reproducibility within a short generation time. The utilization of these aforementioned features is critical in developing sensitive and portable POC systems. Furthermore, the deficiencies related to past experimental efforts to improve biosensor schematics, including the design of biorecognition elements, can be tackled with the integration of computational tools. These complementary tools enable the prediction of the reliability and functionality of the molecular structure of aptamers. In this review, we have overviewed the usage of aptamers in the development of novel and portable POC devices, in addition to highlighting the insights that simulations and other computational methods can provide into the use of aptamer modeling for POC integration.

Development of a Plasmonic Sensor for a Chemotherapeutic Agent Cabazitaxel.

Drug dosage is a crucial subject in both human and animal treatment. Administering less drug dosage may prevent treatment or make it less effective, and high drug dosage may cause a heightened risk of adverse effects, or in some cases, cost a patient's life. Also, even when the dosage is administered carefully, metabolic differences may cause different effects on different patients. Because of these considerations, monitoring drug dosage in the body is a critical and significant requirement in the health industry. Within the scope of this study, a reusable surface plasmon resonance (SPR) chip with fast response, high selectivity, and no pretreatment is produced for the chemotherapeutic agent cabazitaxel. A cabazitaxel-imprinted nanofilm was synthesized on the sensor chip surface and characterized by atomic force microscopy, ellipsometry, and contact angle measurements. Standard cabazitaxel solution and an artificial plasma sample were used for the kinetic analysis. Docetaxel, methylprednisolone, and dexamethasone were analyzed for their selectivity experiment. In addition, the repeatability and storage durability of the sensor were also evaluated. As a result of the adsorption studies, the limit of detection and limit of quantitation values were found to be 0.012 and 0.036 µg/mL, respectively. High-performance liquid chromatography analysis was used to validate the response of the cabazitaxel-imprinted sensor.

A surface plasmon resonance sensor with synthetic receptors decorated on graphene oxide for selective detection of benzylpenicillin.

Antibiotic residues in foods, water and the environment reveal antibiotic-resistant bacterial strains, disrupting the ecological balance and causing serious health problems. For these reasons, the detection of antibiotic residues is crucial for the protection of human health. Herein, the detection of benzylpenicillin antibiotic from aqueous and milk sample solutions was carried out by surface plasmon resonance (SPR) sensor using synthetic receptor-molecularly imprinted polymer. The benzylpenicillin-imprinted poly(hydroxyethyl methacrylate-graphene oxide-N-methacryloyl-l-phenylalanine) (MIP-GO) SPR sensor was prepared. Benzylpenicillin detection was performed by MIP-GO SPR sensor in a 1-100 ppb concentration range of benzylpenicillin with 0.9665 linear correlation and 0.021 ppb detection limit. Selectivity analysis showed that the MIP-GO SPR sensor detected the benzylpenicillin molecule 8.16 times more selectively than amoxicillin and 14.04 times more selectively than ampicillin. To examine the imprinting efficiency, non-imprinted poly(hydroxyethyl methacrylate-graphene oxide-N-methacryloyl-l-phenylalanine) (NIP-GO) SPR sensor was also prepared using the same procedure without benzylpenicillin addition. Since graphene oxide (GO) was added to enhance the sensor signal response by increasing sensitivity, the control analyses were performed by a poly(hydroxyethyl methacrylate-N-methacryloyl-l-phenylalanine) (MIP) SPR sensor without adding GO. Moreover, repeatability studies of MIP-GO SPR sensor were statistically evaluated and the RSD of intra-day assays less than 1% specified that there was no loss of performance for the benzylpenicillin detection ability even after four cycles. As a real food sample analysis, the benzylpenicillin spiked and unspiked milk samples were evaluated and high-performance liquid chromatography experiments were carried out for validation.

Selective Amplification of Plasmonic Sensor Signal for Cortisol Detection Using Gold Nanoparticles.

Herein, gold nanoparticles (AuNP)-modified cortisol-imprinted (AuNP-MIP) plasmonic sensor was developed for signal amplification and real-time cortisol determination in both aqueous and complex solutions. Firstly, the sensor surfaces were modified with 3-(trimethoxylyl)propyl methacrylate and then pre-complex was prepared using the functional monomer N-methacryloyl-L-histidine methyl ester. The monomer solution was made ready for polymerization by adding 2-hydroxyethyl methacrylate to ethylene glycol dimethacrylate. In order to confirm the signal enhancing effect of AuNP, only cortisol-imprinted (MIP) plasmonic sensor was prepared without AuNP. To determine the selectivity efficiency of the imprinting process, the non-imprinted (AuNP-NIP) plasmonic sensor was also prepared without cortisol. The characterization studies of the sensors were performed with atomic force microscopy and contact angle measurements. The kinetic analysis of the AuNP-MIP plasmonic sensor exhibited a high correlation coefficient (R2 = 0.97) for a wide range (0.01-100 ppb) with a low detection limit (0.0087 ppb) for cortisol detection. Moreover, the high imprinting efficiency (k' = 9.67) of the AuNP-MIP plasmonic sensor was determined by comparison with the AuNP-NIP plasmonic sensor. All kinetic results were validated and confirmed by HPLC.

Gold-Modified Molecularly Imprinted N-Methacryloyl-(l)-phenylalanine-containing Electrodes for Electrochemical Detection of Dopamine.

A molecularly imprinted polymer-based pencil graphite electrode (MIP PGE) sensor, modified with gold nanoparticles, was utilized for the detection of dopamine in the presence of other biochemical compounds using cyclic voltammetry (CV) and differential pulse voltammetry (DPV), depending on its strong electroactivity function. The pulse voltammetry methods recorded the highest response. In addition to the high oxidation rate of DA and the other biomolecule interferences available in the sample matrix used, which cause overlapping voltammograms, we aimed to differentiate them in a highly sensitive limit of detection range. The calibration curves for DA were obtained using the CV and DPV over the concentration range of 0.395-3.96 nM in 0.1 M phosphate buffer solution (PBS) at pH 7.4 with a correlation coefficient of 0.996 and a detection limit of 0.193 nM. The electrochemical technique was employed to detect DA molecules quantitatively in human blood plasma selected as real samples without applying any pre-treatment processes. MIP electrodes proved their ability to detect DA with high selectivity, even with epinephrine and norepinephrine competitor molecules and interferences, such as ascorbic acid (AA). The high level of recognition achieved by molecularly imprinted polymers (MIPs) is essential for many biological and pharmaceutical studies.

Selective Detection of Penicillin G Antibiotic in Milk by Molecularly Imprinted Polymer-Based Plasmonic SPR Sensor.

Molecularly imprinted polymer-based surface plasmon resonance sensor prepared using silver nanoparticles was designed for the selective recognition of Penicillin G (PEN-G) antibiotic from both aqueous solution and milk sample. PEN-G imprinted sensors (NpMIPs) SPR sensor was fabricated using poly (2-hydroxyethyl methacrylate-N-methacroyl-(L)-cysteine methyl ester)-silver nanoparticles-N-methacryloyl-L-phenylalanine methyl ester polymer by embedding silver nanoparticles (AgNPs) into the polymeric film structure. In addition, a non-imprinted (NpNIPs) SPR sensor was prepared by utilizing the same polymerization recipe without addition of the PEN-G template molecule to evaluate the imprinting effect. FTIR-ATR spectrophotometer, ellipsometer, contact angle measurements were used for the characterization of NpMIPs SPR sensors. The linear concentration range of 0.01-10 ng/mL PEN-G was studied for kinetic analyses. The augmenting effect of AgNPs used to increase the surface plasmon resonance signal response was examined using polymer-based PEN-G imprinted (MIPs) sensor without the addition of AgNPs. The antibiotic amount present in milk chosen as a real sample was measured by spiking PEN-G into the milk. According to the Scatchard, Langmuir, Freundlich and Langmuir-Freundlich adsorption models, the interaction mechanism was estimated to be compatible with the Langmuir model.

S-citalopram imprinted monolithic columns for capillary electrochromatography enantioseparations.

In this study, the molecular imprinting method was used to separate enantiomeric forms of chiral antidepressant drug, R,S-citalopram (R,S-CIT) in aqueous solution by CEC system combining the advantages of capillary electrophoresis (CE) and high-performance liquid chromatography (HPLC). For that, an amino acid-based molecularly imprinted monolithic capillary column was designed and used as a stationary phase for selective separation of S-citalopram (S-CIT) for the first time. S-CIT was selectively separated from the aqueous solution containing the other enantiomeric form of R-CIT, which is the same in size and shape as the template molecule. Morphology of the molecularly imprinted (MIP S-CIT) and non-imprinted (NIP S-CIT) monolithic capillary columns was observed by scanning electron microscopy. Imprinting efficiency of MIP S-CIT monolithic capillary column used for selective S-CIT separation was verified by comparing with NIP S-CIT and calculated imprinting factor (I.F:1.81) proved the high selectivity of the MIP S-CIT for S-CIT. Cavities formed for S-CIT form enabled selective (α = 2.08) separation of the target molecule from the other enantiomeric R-CIT form. Separation was achieved in a short period of 10 min, with the electrophoretic mobility of 7.68 × 10-6 m2 /Vs for R,S-CIT at pH 7.0 10 mM PB and 50% ACN ratio. The performance of both MIP S-CIT and NIP S-CIT columns was estimated by repeating the R,S-CIT separations with intra-batch and inter-batch studies for reproducibility of retention times of R,S-CITs. Estimated RSD values that are lower than 2% suggest that the monolithic columns separate R,S-CIT enantiomers without losing separation efficiency.

Sensitive and selective detection of amitrole based on molecularly imprinted nanosensor.

SPR sensor used for amitrole detection was prepared without using any modification. Molecularly imprinted SPR sensor enabled high selectivity for amitrole pesticide. Amino acid-based functional monomer MATrp was integrated as a recognition element. Tailor-made SPR sensor enables real-time monitoring of amitrole pesticide. Synthetic recognition sites provided by MATrp were prepared without labeling.

Selective recognition of nucleosides by boronate affinity organic-inorganic hybrid monolithic column.

Boronic acids are important ligands used to selectively recognize and enrich compounds containing cis-diol groups such as nucleosides. In the present study, organic-inorganic hybrid [POSS-MAH-BPA] monolithic column was prepared for the first time in the literature as a new boronate affinity system for the recognition of nucleosides. The selectivity of the [POSS-MAH-PBA] boronate affinity monolithic column for the recognition of cis-diol containing adenosine nucleoside from its analogue molecule of deoxyadenosine triphosphate, dATP, non-cis-diol containing compound was investigated both by UV and HPLC studies. When the relative selectivity coefficients are compared, the [POSS-MAH-PBA] boronate affinity monolithic column is 4.25 times more selective for adenosine than [POSS-MAH] monolithic column. Besides, to determine endogenous nucleosides in biological fluids, which may serve as non-invasive cancer biomarkers, nucleosides were spiked into the urine solutions and passed through the [POSS-MAH-PBA] boronate affinity monolithic column, and the nucleosides were confirmed by HPLC. The adenosine recognition capability of the [POSS-MAH-PBA] boronate affinity monolithic column with an average enrichment factor of 48.9-fold was apparently superior to that of the [POSS-MAH] monolithic column. Methacryl Polyhedral Oligomeric Silsesquioxanes (POSS-MA) with nano-sized stable 3-dimensional architectures provided the advantage of being used as an adsorbent for the monolithic structure by providing high surface area, 507.60 m2/g, and enabling vinyl groups to function with amino acid-based MAH monomers capable of providing electrons to coordinate PBA. Recovery results of more than 90% for adenosine showed that the [POSS-MAH-PBA] boronate affinity monolithic column could be a promising adsorbent for selective adsorption of cis-diol containing compounds such as nucleosides.

Molecularly imprinted cryogel cartridges for the selective recognition of tyrosine.

Molecularly imprinted polymers are used for creating a specific cavity and selective recognition sites for the structure of a target molecule in a polymeric structure. In this study, specific molecularly imprinted cryogel cartridges were synthesized using two distinct functional monomers to compare imprinting efficiency for the selective recognition of Tyrosine (Tyr). Tyr-imprinted cryogel cartridge (MIP1) was prepared using metal-chelate coordination for the imprinting process by free-radical bulk polymerization under frozen conditions, and Tyr-imprinted cryogel cartridge (MIP2) was prepared in the same way using hydrophobic effects for imprinting. After the characterization of the cryogel cartridges was carried out, the optimum adsorption conditions of both were determined according to the different parameters such as flow rate (0.5-2.5 ml/min), pH of the medium (4.0-8.0), initial Tyr concentration (0.1-3.0 mg/ml), and temperature (4-45°C). Selectivity experiments of Tyr-imprinted and non-imprinted cryogel cartridges were carried out by using phenylalanine, tryptophan, and cysteine. Besides, the eluted Tyr from MIP1 and MIP2 cryogel cartridge were applied to FPLC system. Also, the reusability experiments of Tyr-imprinted cryogel cartridges was observed no significant decrease in the adsorption capacity.

A dye-affinity cryogel membrane for malate dehydrogenase purification from Saccharomyces cerevisiae.

Cibacron blue F3GA functionalized poly(hydroxyethyl methacrylate) cryogel membranes were prepared and applied for a simple purification of malate dehydrogenase (MDH) from crude extract of Saccharomyces cerevisiae. Swelling tests, scanning electron microscopy, surface area measurements and Fourier transform infrared (FTIR) spectroscopy techniques were used for the characterization of dye-affinity cryogel membranes. Following cell homogenization and extraction, MDH was purified using the dye-affinity cryogel membranes at a high yield of 80.5% with 54-fold purification. Maximum MDH adsorption amount was determined to be 267.7 mg/g of membranes at pH 7.4, 25 °C and a flow rate of 1.0 mL/min. Interestingly, the cryogel membranes were used for several purification runs without any significant decrease in MDH adsorption capacity. Sodium dodecyl sulfate polyacrylamide gel electrophoresis was carried out to assess the purity of the eluted MDH. The obtained results highlight the dye-affinity cryogel membranes as powerful dye affinity adsorbents for MDH purification from S. cerevisiae.

Phenol removal from wastewater by surface imprinted bacterial cellulose nanofibres.

In this study, we have reported a novel wastewater treatment technique by phenol imprinted bacterial cellulose (BC-MIP) nanofibres with high specificity and adsorption capacity. N-methacryloyl-(L) phenylalanine methyl ester (MAPA) functional monomer was used to create specific binding sites for the template molecule phenol via electrostatic and hydrophobic interactions. BC-MIP nanofibres were synthesized by surface imprinting approach in the presence of different amounts of total monomer (% weight), monomer/template ratio and polymerization time. Then, the nanofibres were characterized by FTIR-ATR, surface area analysis (BET), elemental analysis, scanning electron microscopy (SEM) and contact angle measurements. Adsorption studies were performed with respect to pH, temperature and ionic strength, and the adsorption capacity was calculated by using the spectrophotometer. In order to desorb the adsorbed phenol from BC-MIP nanofibres, 0.1 M NaCl solution was used. Besides, BC-MIP nanofibres were applied to real wastewater samples from Ergene basin in Turkey. The suitable equilibrium isotherm was determined as Langmuir isotherm. To evaluate the selectivity of the BC-MIP nanofibres, similar molecules were utilized as competitor molecules, which were 2-chlorophenol, 4-chlorophenol and 2,4-dichlorophenol. Electrostatic interactions were found to contribute to the generation of specific recognition binding sites. The results have shown that imprinting of phenol was achieved successfully with high adsorption capacity. The phenol removal efficiency was reported up to 97%. BC-MIP nanofibres were used 10 times with a negligible decrease in adsorption capacity.

Polymethacryloyl-L-Phenylalanine [PMAPA]-Based Monolithic Column for Capillary Electrochromatography.

The ability to detect catecholamines (CAs) and their metabolites is vital to understand the mechanism behind the neuronal diseases. Neurochemistry aims to provide an improved pharmacological, molecular and physiological understanding of complex brain chemistries by analytical techniques. Capillary electrophoresis (CE) is one such analytical technique that enables the study of various chemical species ranging from amino acids and peptides to natural products and drugs. CE can easily adapt the changes in research focus and in recent years remains an applicable technique for investigating neuroscience and single cell neurobiology. The prepared phenylalanine-based hydrophobic monolithic column, Polymethacryloyl-L-phenylalanine [PMAPA], was used as a stationary phase in capillary electrochromatography to separate CAs that are similar in size and shape to each other including dopamine (DA) and norepinephrine (NE) via hydrophobic interactions. Separation carried out in a short period of 17 min was performed with the electrophoretic mobility of 5.54 × 10-6 m2 V-1 s-1 and 7.60 × 10-6 m2 V-1 s-1 for DA and NE, respectively, at pH 7.0, 65% acetonitrile ratio with 100 mbar applied pressure by the developed hydrophobic monolithic column without needing any extra process such as imprinting or spacer arms to immobilize ligands used in separation.

Protein depletion with bacterial cellulose nanofibers.

In this study, we have reported a novel fabrication technique for human serum albumin (HSA) imprinted composite bacterial cellulose nanofibers (MIP-cBCNFs) used for the depletion of HSA selectively from artificial blood plasma for proteomic applications. Molecular imprinting was achieved by using metal ion coordination interactions of N­methacryloyl­(l)­histidinemethylester (MAH) monomer and Cu(II) ions. MAH-Cu(II)-HSA complex was polymerized with bacterial cellulose nanofibers (BCNFs) under constant stirring at room temperature. The characterization of the MIP-cBCNFs was carried out by FTIR-ATR, SEM, contact angle measurements and surface area measurements. The adsorption experiments of HSA onto the MIP-BCNFs and NIP-BCNFs from aqueous HSA solutions were investigated in a batch system. The selectivity of the MIP-cBCNFs was investigated by using non-template human transferrin (HTR), and myoglobin (Myo). The relative selectivity coefficients of the MIP-cBCNFs were calculated as 4.73 and 3.02 for HSA/HTR and HSA/Myo molecules, respectively. In addition, the depletion of HSA from artificial human plasma was confirmed by SDS-PAGE and 2-D gel electrophoresis. As a result, it has been shown that metal ion coordination interactions contribute to specific binding of template when preparing MIP-cBCNFs for the depletion of HSA with a high adsorption capacity, significant selectivity and reusability.

Catalase purification from rat liver with iron-chelated poly(hydroxyethyl methacrylate-N-methacryloyl-(l)-glutamic acid) cryogel discs.

In this study, iron-chelated poly(hydroxyethyl methacrylate-N-methacryloyl-(l)-glutamic acid) (PHEMAGA/Fe(3+)) cryogel discs were prepared. The PHEMAGA/Fe(3+) cryogel discs were characterized by elemental analysis, scanning electron microscopy, Fourier transform infrared spectroscopy, swelling tests, and surface area measurements. The PHEMAGA/Fe(3+) cryogel discs had large pores ranging from 10 to 100 µm with a swelling degree of 9.36 g H2O/g cryogel. Effects of pH, temperature, initial catalase concentration, and flow rate on adsorption capacity of the PHEMAGA/Fe(3+) cryogel discs were investigated. Maximum catalase adsorption capacity (62.6 mg/g) was obtained at pH 7.0, 25°C, and 3 mg/ml initial catalase concentration. The PHEMAGA/Fe(3+) cryogel discs were also tested for the purification of catalase from rat liver. After tissue homogenization, purification of catalase was performed using the PHEMAGA/Fe(3+) cryogel discs and catalase was obtained with a yield of 54.34 and 16.67 purification fold.

Synthesis of a specific monolithic column with artificial recognition sites for L-glutamic acid via cryo-crosslinking of imprinted nanoparticles.

In this study, a new molecular imprinting (MIP)-based monolithic cryogel column was prepared using chemically crosslinked molecularly imprinted nanoparticles, to achieve a simplified chromatographic separation (SPE) for a model compound, L-glutamic acid (L-Glu). Cryogelation through crosslinking of imprinted nanoparticles forms stable monolithic cryogel columns. This technique reduces the leakage of nanoparticles and increases the surface area, while protecting the structural features of the cryogel for stable and efficient recognition of the template molecule. A non-imprinted monolithic cryogel column (NIP) was also prepared, using non-imprinted nanoparticles produced without the addition of L-Glu during polymerization. The molecularly imprinted monolithic cryogel column (MIP) indicates apparent recognition selectivity and a good adsorption capacity compared to the NIP. Also, we have achieved a significant increase in the adsorption capacity, using the advantage of high surface area of the nanoparticles.

Removal of iron by chelation with molecularly imprinted supermacroporous cryogel.

Iron chelation therapy can be used for the selective removal of Fe(3+) ions from spiked human plasma by ion imprinting. N-Methacryloyl-(L)-glutamic acid (MAGA) was chosen as the chelating monomer. In the first step, MAGA was complexed with the Fe(3+) ions to prepare the precomplex, and then the ion-imprinted poly(hydroxyethyl methacrylate-N-methacryloyl-(L)-glutamic acid) [PHEMAGA-Fe(3+)] cryogel column was prepared by cryo-polymerization under a semi-frozen temperature of - 12°C for 24 h. Subsequently, the template, of Fe(3+) ions was removed from the matrix by using 0.1 M EDTA solution. The values for the specific surface area of the imprinted PHEMAGA-Fe(3+) and non-imprinted PHEMAGA cryogel were 45.74 and 7.52 m(2)/g respectively, with a pore size in the range of 50-200 µm in diameter. The maximum Fe(3+) adsorption capacity was 19.8 µmol Fe(3+)/g cryogel from aqueous solutions and 12.28 µmol Fe(3+)/g cryogel from spiked human plasma. The relative selectivity coefficients of ion-imprinted cryogel for Fe(3+)/Ni(2+) and Fe(3+)/Cd(2+) were 1.6 and 4.2-fold greater than the non-imprinted matrix, respectively. It means that the PHEMAGA-Fe(3+) cryogel possesses high selectivity to Fe(3+) ions, and could be used many times without significantly decreasing the adsorption capacity.

Characterization and cellular interaction of fluorescent-labeled PHEMA nanoparticles.

The aim of this study is to label the cells with polymeric nanoparticles properly and efficiently. For this purpose, acridine orange (AO)-loaded poly(2-hydroxyethyl methacrylate) (PHEMA) nanoparticles were synthesized by miniemulsion polymerization. PHEMA nanoparticles were characterized by zeta sizer, zeta potential, atomic force microscopy (AFM), scanning electron microscopy (SEM), transmission electron microscopy (TEM), fluorescence microscopy, Fourier transform infrared spectrophotometer (FTIR), and elemental analysis. In addition, the toxicity of the nanoparticles were investigated by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay.