RESUMO
A rapid and simple method of biofunctionalising nylon, cellulose acetate, and polyvinyl butyral electrospun nanofibers with blood group glycans was achieved by preparing function-spacer-lipid constructs and simply contacting them to fibers with a piezo inkjet printer. A series of water dispersible amphipathic glycan-spacer constructs were synthesized representing a range ABO and related blood group antigens. After immediate contact of the amphipathic glycan-spacer constructs with nanofiber surfaces they self-assembled and were detectable by enzyme immunoassays with high sensitivity and specificity.
Assuntos
Sistema ABO de Grupos Sanguíneos/química , Carboidratos/química , Nanofibras/química , HumanosRESUMO
A prerequisite for exploiting sensing devices based on semiconductor nanowires is ultra-sensitive and selective direct electrical detection of biological and chemical species. Here, we constructed a transducer based on copolymer of poly(3,4,-ethylenedioxythiophene) (PEDOT) and carboxylic group functionalised PEDOT single nanowire in between gold electrodes, followed by covalent attachment of amino-modified probe oligonucleotide. The target ODNs specific to Homo sapiens Breast and ovarian cancer cells were detected at femtomolar concentration and incorporation of negative controls (non-complementary ODN) were clearly discriminated by the sensor. The ex situ measurements were performed by using two terminal device setup and the changes in the interface of the nanowire associated with the association or dissociation of ODNs were measured as change in resistance. In addition, in situ measurements were performed by utilizing scanning ion conductance microscopy to measure the change in resistance of probe modified nanowire upon addition of different concentration of target ODNs in presence of relevant buffer. The constructed, nano sensor showed highly sensitive concentration dependent resistance change.
Assuntos
Técnicas Biossensoriais/métodos , DNA de Neoplasias/análise , DNA de Neoplasias/genética , Genes BRCA1 , Nanofios , Sequência de Bases , Técnicas Biossensoriais/instrumentação , Técnicas Biossensoriais/estatística & dados numéricos , Neoplasias da Mama/genética , Compostos Bicíclicos Heterocíclicos com Pontes , Feminino , Humanos , Nanofios/química , Sondas de Oligonucleotídeos/genética , Neoplasias Ovarianas/genética , Polímeros , Pontos Quânticos , Sensibilidade e EspecificidadeRESUMO
Label-free oligonucleotide sensors that use a change in the electrode kinetics of the redox reaction of the negatively charged Fe(CN)(6)(3-/4-) redox couple to signal the formation of a DNA duplex with a surface-conjugated probe nucleotide are investigated. Electrochemically active conducting polymers (ECPs) can advantageously be used both as the active electrode and as the means of surface conjugation of the probe nucleotide. Here, we demonstrate that the sensitivity of the detection of the surface-complementary oligonucleotide can significantly be improved, into the low nanomolar range, by forming the ECP as a highly porous, very rough layer by growing it using electrochemical polymerization on a microelectrode. In comparison, smoother surfaces formed on macroelectrodes had detection sensitivity in the low micromolar range. We propose Donnan exclusion of the redox couple from small pores as the reason for the enhanced sensitivity. We discuss the effects using a simple patch model for the electrochemical kinetics and use the model to derive the equilibrium binding constant and binding kinetic rate constants for the surface hybridization reaction. We use the electrochemically active copolymer of pyrrole (Py) and 3-pyrrolylacrylic acid (PAA) [poly(Py-co-PAA)] as the sensing electrode and binding surface and measure the surface hybridization-induced changes in electrode kinetics of Fe(CN)(6)(3-/4-) by electrochemical impedance spectroscopy.