Application of HRP-streptavidin bionanoparticles for potentiometric biotin determination.

Biotin is widely used in infant formula to prevent biotin deficiency of newborn babies and in beauty products as nutritional supplements for coenzymatic functions and having strong nails, shiny hair, and skin over the last few years. There is a need for the development of a fast, simple and reusable assay method to perform biotin determination at very low concentrations. Biotin determination has achieved with a prepared potentiometric biotin sensor that has a very wide concentration range (10-15M-10-7M) and a lower detection limit (0.3 10-15M) with a very good regression coefficient (0.9925). A quick response (7 min), good accuracy (recovery 100.4-103.7%), reproducible, reusable (10 times), and long-term stability (3 months) have been obtained using the prepared potentiometric sensor. The obtained results have proved that the prepared potentiometric sensor can be used for biotin determination in real samples.

Proteinous Polymeric Shell Decorated Nanocrystals for the Recognition of Immunoglobulin M.

This study demonstrates the preparation of photosensitively orientated and crosslinked proteinous polymeric shell having quantum dot based nanocrystals through Amino acid Decorated and Light Underpinning Conjugation Approach (ANADOLUCA). ANADOLUCA is based on photo-electron transfer method and uses these decorated nanocrystals for specifically and effectively recognition and detection of Immunoglobulin M in the aqueous environment. The conjugation method effectively provides an orientation of affinity pairs on the surface of quantum dots nanocrystals. This photosensitive ruthenium-based amino acid monomer is a synthetic and inexpensive material for the preparation of bioconjugates. The nanocrystals give advantages for using a wide pH and temperature range. The construction and preparation method is applicable to silica materials, superparamagnetic particles, quantum dots, carbon nanotubes, Ag/Au nanoparticles, Au surfaces, and polymeric materials. This prepared proteinous polymeric shell decorated nanocrystals are of great potential in applications in life sciences and can be used in infection case studies or allergy symptoms.

Long-term stability and reusability of molecularly imprinted polymers.

Molecularly imprinted materials are man-made mimics of biological receptors. Their polymer network has recognition sites complementary to a substrate in terms of size, shape and chemical functionality. They have diverse applications in various chemical, biomedical and engineering fields such as solid phase extraction, catalysis, drug delivery, pharmaceutical purification, (bio)sensors, water treatment, membrane separations and proteomics. The stability and reusability of molecularly imprinted polymers (IPs) have crucial roles in developing applications that are reliable, economic and sustainable. In the present article the effect of crosslinkers, functional monomers and conditions for template extraction on the long-term stability and reusability of IPs was systematically investigated. Adsorption capacity, selectivity, morphology and thermal decomposition of eleven different l-phenylalanine methyl ester imprinted polymers were studied to reveal performance loss over 100 adsorption-regeneration cycles. Furthermore, crosslinker and functional monomer specific reversible and irreversible decomposition of imprinted polymers as a result of adsorbent regeneration were investigated through adsorption studies, electron microscopy, N2 adsorption and thermogravimetric analysis. A decomposition mechanism was proposed and revealed using NMR spectroscopy. Solutions to avoid or overcome the limitations of the most common crosslinkers, functional monomers and extraction techniques were proposed and experimentally validated.

Phosphoserine imprinted nanosensor for detection of Cancer Antigen 125.

Cancer antigen 125 (CA 125) is widely used as diagnostic biomarker for ovarian cancer. Change in the concentration level of CA 125 is associated with disease progression or regression. CA 125 posseses a phosphorylation site and protein backbone is phosphorylated on serine, before secretion. In this study, we have developed an imprinting method for CA 125 recognition and determination. In this method, methacryloyl antipyrine europium (III) [(MAAP)2-Eu(III)] and methacryloyl antipyrine terbium (III) [(MAAP)2-Tb(III)] have been used as new metal-chelating monomers via metal coordination-chelation interactions. Phosphoserine (PS) has been used as a template for the detection of CA 125. PS imprinted carbon nanotube (CNT) and Fe2O3 nanoparticle (SPN) have cavities that are selective for CA 125. The binding affinity of the PS imprinted CNT and SPN nanosensor has been investigated using Langmuir adsorption isotherms and affinity constants (Kaffinity) have found to be 1.85 105M-1 for PS and 13.5 10-3 mLU-1 and 7.73 10-3 mLU-1 for CA 125 (for CNT and SPN, respectively). Detection limit of PS imprinted CNT nanosensor for PS and CA 125 have been found to be 1.77 10-10M and 0.49 UmL-1, respectively. Human serum samples have been spiked with different concentrations of CA 125 (in pH 7.4 PBS) to investigate the feasibility of the nanosensors for clinical applications. Experimental results have been revealed that prepared nanosensors have been exhibited better sensivity, recovery and reproducibility.

Preparation and characterization of methotrexate-loaded microcapsules.

Methotrexate (MTX) has toxic effect to healthy tissues. Microencapsulation coats particles with a functional coat to optimize storage stability and to modulate release. In the present study, a new MTX encapsulated microcapsules were synthesized for controlling MTX release. Controlled drug release provides releasing of efficient dose and prevent drug side effect to tissues and also protects MTX from oxygen, pH and other interactions. MTX was encapsulated through biocompatible hyaluronic acid (HA) and sodium alginate (SA) with an encapsulation system to reduce its toxicity and for controlled release. The microcapsules prepared by vibrating nozzle were cross-linked with SA, HA and calcium chloride. Nozzle diameter and MTX concentration were changed according to loaded MTX and encapsulation efficiency were determined using HPLC. For the reliability of the data, validation studies of the HPLC method were performed. The precision of the method was demonstrated using intra- and inter-day assay relative standard deviation (RSD) values which are less than 2% in all instances. For the characterization of microcapsules, particle size, drug loading and in vitro drug release studies were performed. Diameters of MTX-loaded microcapsules were acquired approximately 160, 400 and 800 µm. Surface morphology of encapsulated microcapsules were displayed with light microscope. Eighty-nine percent MTX encapsulation efficiencies were achieved. Encapsulated MTX microcapsules showed controlled release when compared to pure MTX. While powder MTX dissolved completely in 10 min in the dissolution medium, MTX release from encapsulated MTX microcapsules became 40 h in 0.1 M PBS pH 7.4, including NaCl. MTX release from MTX-loaded microcapsules was reached to 79%. Moreover, drug efficiency was examined in vitro cell culture tests. Viability of 5RP7 cells were decreased to 88.5% for 96 h. When MTX was given directly to 5RP7 cells, viability of 5RP7 cells was decreased to 49.7% for 96 h. Flow cytometry studies also showed that, MTX microcapsules induced apoptosis. The goal of this study is to provide controlled release of MTX and to reduce the toxic effect of MTX.

Mutual recognition of TNT using antibodies polymeric shell having CdS.

Click chemistry is the latest strategy called upon in the development of state of the art exponents of bioconjugation. In this study, we have proposed a covalent and photosensitive crosslinking conjugation of the antibody on nano-structures. For this purpose, quantum dots (QDs) without affecting conformation and function of proteins through the ruthenium-chelate based aminoacid monomer linkages have been applied. The aminoacid-monomer linkages called ANADOLUCA (AmiNoAcid Decorated and Light Underpining Conjugation Approach) give reusable oriented and cross-linked anti 2,4,6-trinitrotoluene (TNT) conjugated QD for TNT detection. In this work, a new and simple method has improved to design and prepare high sensitive nanoconjugates for TNT determination. We have demonstrated the use of luminescent QDs conjugated to antibody for the specific detection of the explosive TNT in aqueous environments. The binding affinity of each nanoconjugates for TNT detection by using Langmuir adsorption methods has also been investigated.

Quantum dot nanocrystals having guanosine imprinted nanoshell for DNA recognition.

Molecular imprinted polymers (MIPs) as a recognition element for sensors are increasingly of interest and MIP nanoparticles have started to appear in the literature. In this study, we have proposed a novel thiol ligand-capping method with polymerizable methacryloylamido-cysteine (MAC) attached to CdS quantum dots (QDs), reminiscent of a self-assembled monolayer and have reconstructed surface shell by synthetic host polymers based on molecular imprinting method for DNA recognition. In this method, methacryloylamidohistidine-platinium (MAH-Pt(II)) is used as a new metal-chelating monomer via metal coordination-chelation interactions and guanosine templates of DNA. Nanoshell sensors with guanosine templates give a cavity that is selective for guanosine and its analogues. The guanosine can simultaneously chelate to Pt(II) metal ion and fit into the shape-selective cavity. Thus, the interaction between Pt(II) ion and free coordination spheres has an effect on the binding ability of the CdS QD nanosensor. The binding affinity of the guanosine imprinted nanocrystals has investigated by using the Langmuir and Scatchard methods, and experiments have shown the shape-selective cavity formation with O6 and N7 of a guanosine nucleotide (K(a) = 4.841x10(6) mol L(-1)) and a free guanine base (K(a) = 0.894x10(6) mol L(-1)). Additionally, the guanosine template of the nanocrystals is more favored for single stranded DNA compared to double stranded DNA.

Mimicking receptor for methylmercury preconcentration based on ion-imprinting.

Solid-phase extraction (SPE) based on molecularly imprinted polymers (MIPs) were used to develop selective separation and preconcentration for methylmercury ion from complex matrixes. In this study, an ion-imprinting polymer was prepared to make artificial organomercury lyase preorganizing three methacryloyl-(L)-cysteine methylester (MAC) monomers and one methylmercury ion in a three-coordinate form by template polymerization, with the goal preparing a solid-phase which has the high selectivity for methylmercury ions. Methylmercury-imprinted beads were produced by a dispersion polymerization technique with use methylmercury-methacryloyl-(L)-cysteine (MM-MAC) complex monomer and ethylene glycoldimethacrylate (EDMA). After removal of methylmercury ions, methylmercury-imprinted beads were used for solid-phase extraction and determination of mercury compounds. Methylmercury adsorption and selectivity studies of methylmercury versus other metal ions which Hg(II), Zn(II), Pb(II) and Cd(II) were reported and distribution and selectivity coefficients of these ions with respect to methylmercury were calculated here. ICP-OES and HPLC-DAD determinations of methylmercury and mercury ions in the certified reference, LUTS-1 from the National Research Council of Canada and synthetic sea water showed that the interfering matrix had been almost removed during preconcentration. The methylmercury-imprinted solid-phase as mimic receptor was good enough for methylmercury determination in complex matrixes.

Selective preconcentration of thorium in the presence of UO(2)(2+), Ce(3+) and La(3+) using Th(IV)-imprinted polymer.

We have prepared Th(IV) ion-imprinted polymers, which can be used for the selective preconcentration of Th(IV) ions, represented by uranium and lanthanides. N-methacryloyl-(l)-glutamic acid (MAGA) was chosen as the complexing monomer. In the first step, Th(IV) was complexed with MAGA and the Th(IV)-imprinted poly[ethylene glycol dimethacrylate-N-methacryloyl-(l)-glutamic acid] (Poly(MAGA-EDMA)) beads were synthesized by suspension polymerization. After that, the template Th(IV) ions were removed using 8.0M HNO(3) solution. The breakthrough capacity was 40.44mg Th(IV)/g beads. The relative selectivity coefficients of imprinted beads were 68, 97 and 116 for UO(2)(2+), La(3+) and Ce(3+), times greater than non-imprinted matrix, respectively. The Th(IV)-imprinted beads could be used many times without decreasing their breakthrough capacities significantly.