ABSTRACT
Hybrid thin films based on Hydrocalumite (Ca2AlCl layered double hydroxide LDH) and tyrosinaseenzyme have been used for the elaboration of a high sensitive amperometric biosensor detecting polyphenols extracted from green tea. Structural properties of LDH nanomaterials were characterized by X-ray powder diffraction and Infra-Red spectroscopy, confirming its crystalline phase and chemical composition. Ca2AlCl-LDHs-thin films were deposited by spin-coating, and studied by atomic force microscopy to obtain information about the surface morphology of this host matrix before and after enzyme's immobilization. Electrochemical study using cyclic voltammetry and chronoamperometry shows good performances of the built-in biosensor with a high sensitivity for polyphenols concentrations ranging from 24 pM to and a limit of detection of 1.2 pM.
Subject(s)
Aluminum Oxide/chemistry , Biosensing Techniques/methods , Calcium Chloride/chemistry , Electrochemical Techniques/methods , Polyphenols/analysis , Biosensing Techniques/instrumentation , Electrochemical Techniques/instrumentation , Enzymes, Immobilized/chemistry , Enzymes, Immobilized/metabolism , Equipment Design , Microscopy, Atomic Force , Monophenol Monooxygenase/chemistry , Monophenol Monooxygenase/metabolism , Nanostructures/chemistryABSTRACT
As a potential biomarker for the investigation of cancer inflammatory profiles, macrophage mannose receptor (MMR, CD206) is herein selected to develop an immunosensor based on layered double hydroxide (LDH). Like an endocyte C-type lectin receptor, MMR plays an important role in immune homeostasis by scavenging unwanted mannose glycoproteins. It attracts a progressive attention thanks to its particularly high expression within the tumor microenvironment. There is a great of interest to develop an immunosensor based on an antibody specific to MMR for detection of stroma versus tumor cells. In this work, we studied the feasibility of high sensitive MMR cancer Screen Printed Electrode (SPE) immunosensor. Working electrode of commercialized SPE was modified by immobilization of specific antibody (anti-MMR) into thin layer of LDH nanomaterials. Structural, morphological, and surface properties of LDHs were studied by X-Ray diffraction, atomic force microscopy and Infrared spectroscopy in ATR. Cyclic Voltammetry technique was used to study interaction between the human recombinant MMR protein (rHu-MMR, NSO derived) and an immobilized antibody into developed immunosensor. High specific response of -11.72 µA/ng.mL(-1) (with a correlation coefficient of R(2)=0.994 ) were obtained in linear range of 0.05 ng/mL to 10.0 ng/mL of specific recombinant antigen. The limit of detection (LOD) was less than 15.0 pg/mL. From these attractive results, the feasibility of an electrochemical immunosensor for cancer was proved. Additional experiments to study stability and reproducibility the immunosensor should be completed in perspective to use these anti-MMR based immunosensors for sensing human MMR in patient biopsies and sera.
Subject(s)
Biomarkers, Tumor/analysis , Conductometry/instrumentation , Hydroxides/chemistry , Immunoassay/instrumentation , Lectins, C-Type/analysis , Mannose-Binding Lectins/analysis , Neoplasms, Experimental/chemistry , Receptors, Cell Surface/analysis , Biomarkers, Tumor/immunology , Equipment Design , Equipment Failure Analysis , Feasibility Studies , Humans , Lectins, C-Type/immunology , Mannose Receptor , Mannose-Binding Lectins/immunology , Neoplasms, Experimental/diagnosis , Neoplasms, Experimental/immunology , Receptors, Cell Surface/immunology , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
This work describes the use of layered double hydroxides (LDHs) for the immobilisation of acetylcholinesterase (AChE) on insulator/semiconductor solid supports. Different LDHs have been synthesised by a co-precipitation method. Afterwards, biohybrid materials based on AChE-LDH mixtures have been produced using wild and recombinant enzymes. Spectroscopic techniques have confirmed the LDH phase identity and the links created between the LDH and AChE. Spectrophotometric assays have demonstrated that most of the biohybrid materials are functional and stable. Several configurations have been used for AChE immobilisation. The highest catalytic responses have been observed when using wild enzyme and immobilising AChE-LDH mixtures on LDHs previously deposited on the solid supports. LDHs have been demonstrated to be suitable host matrices for AChE immobilisation on electrodes for the subsequent development of electrochemical biosensors.
Subject(s)
Acetylcholinesterase/chemistry , Acetylcholinesterase/metabolism , Hydroxides/chemistry , Chemical Precipitation , Chlorides/chemistry , Enzymes, Immobilized/chemistry , Enzymes, Immobilized/metabolism , SpectrophotometryABSTRACT
In this work, we present the development of a hybrid biomembrane based on the immobilization of diamine oxidase (DAO) into LDH thin films for histamine detection. The LDHs preselected as host matrixes are: hydrotalcites (Mg2Al(CO3)0.5(OH)6), lowaite (Mg4Fe(OH)10Cl) and hydrocalumite (Ca2Al(OH)6Cl). The immobilized probes were characterized by atomic force microscopy (AFM) and attenuated total reflection infrared spectroscopy (IR-ATR mode). The analysis of these results shows that the immobilization of DAO occurs with all type of selected LDH and is stable after a 7 day-immersion in phosphate buffer solution. The LDH incorporating magnesium or calcium divalent cations present high-quality surface topology for DAO immobilization and the ability to keep the enzyme in a well conformation for biogenic amines catabolism and histamine detection.
