Application of biosensors in early detection of contamination with lactic acid bacteria during apple juice and concentrate production

The aim of Collective Research Project QUALI-JUICE (COLL-CT-2005, Co Nr 012461) was to introduce the biosensors in control of microbiologically pure apple juice production. Three commercial lactate biosensors were used and compared with enzyme kits assays. Parallel the microbiological analysis of the production process at one juice producing enterprise was done. The results of lactic acid assay with biosensors were in good correlation with those obtained by enzyme kits. The main benefit of biosensor use was shortening of measurement time as compared with assay by enzyme kit and possibility to measure at line. The concentration of l-lactic acid in apple pulp could be correlated with the number of lactic acid bacteria. Pasteurization process was eliminating lactic acid bacteria and the concentration of lactic acid was at this stage not exceeding 0.1 g L⁻¹. The final product (apple concentrate) contained in some cases very high amounts of lactic acid indicating secondary microbiological contamination after pasteurization step. Parallel microbiological analysis of production process and lactate assay indicated that the critical point during production was prolonged vacuum filtration after pasteurization.     

Comparative analysis of QCM and SPR techniques for the optimization of immobilization sequences

One of the most remarkable procedures to immobilize some biological molecules onto surfaces is the use of self-assembled monolayers (SAMs). The aim of this work is to analyse the influence of formation conditions in the detection capability of two different SAMs. With this purpose two techniques have been implemented: the Quartz Crystal Microbalance with Dissipation (QCM-D) and the Surface Plasmon Resonance (SPR). Thus, several parameters usually involved in the SAM protocols have been characterized, i.e. the nature of the thiolated acid. The influence of its concentration and incubation time has been also taken into account. For the validation of these biological layers, the polymyxin B sulfate salt (PmB), as ligand, and the lipopolysaccharide (LPS), as analyte, have been used. It is demonstrated that both in the QCM and the SPR, the use of SAM improves significantly the detection and immobilization of the target compound and an optimum SAM formation protocol is provided.     

Thermodynamic analysis of the interaction between 3-aminophenylboronic acid and monosaccharides for development of biosensor

The special interaction between 3-aminophenylboronic acid and monosaccharides, particularly glucose, fructose and galactose, has been investigated and used for the development of electrochemical and surface plasmon resonance (SPR) based saccharide sensors. The binding constants and thermodynamic parameters of interaction such as free energy of binding (ΔG), enthalpy (ΔH) and entropy (ΔS) were determined for the first time using isothermal titration calorimetry revealed the enthalpic and entropic contributions to the free energies of binding. Effects of pH and buffer on the binding constants (k) between 3-aminophenylboronic acid and monosaccharides were also examined. The binding constant of fructose was found to be six times higher than glucose and four times higher than galactose at pH 11, leading the specific detection of fructose. SPR and potentiometric calibration for fructose were linear in the concentration range of 10–120 mM and 8–84 mM, respectively. Limit of detection of the SPR and potentiometric method were found to be as 8.9 mM and 1.4 mM, respectively.     

Design of an integrated low-noise read-out system for DNA capacitive sensors

This paper presents an electronic system for a fast DNA label-less detection. The sensitivity of the capacitive sensor in use is improved by depositing an insulating self-assembled monolayer (SAM) over the golden electrodes. The capacitance shift due to the hybridization effect is monitored by means of a charge-sensitive amplifier and digitalized by means of a comparator and a counter. The read-out solution demonstrates the ability to identify a 0.01% variation on the capacitive value of the sensor. Results from measurements with the optimized sensor show the reliability of the electronics. The investigated solution is suitable for monolithic systems or for a micro-fabricated array of sensors. An example of the integrated front-end is described and performances and noise evaluation are reported here.     

Amperometric Cholesterol Biosensor Using Layer-by-Layer Adsorption Technique on Polyaniline-coated Polyester Films

An amperometric cholesterol biosensor was fabricated using polyaniline-coated polyester films. Polyaniline was dissolved in chloroform with camphorsulfonic acid, and polystyrene was added to this solution. Using this mixed solution, the coating was placed onto polyester films. Cholesterol oxidase was immobilized onto these films using an electrostatic layer-by-layer adsorption technique. Poly(diallyldimethylammonium chloride) was used as the counter ion source. The level of adsorption was examined and evidence of layer-by-layer adsorption was investigated using a quartz crystal microbalance (QCM). A cholesterol biosensor was fabricated from these films as a working electrode, and it was used to measure the cholesterol concentration.     

Synthesis and Characterization of Sulfonated Polyanilines and Application in Construction of a Diazinon Biosensor

A series of sulfonated polyaniline/derivatized polyaniline nanocomposites was chemically synthesized in the presence of anthracene and naphthalene sulfonic acids. UV-vis and FTIR results indicated the emergence of new bands at 420 and 700 nm and 1100 cm^?1 in their spectra, respectively, meaning the dopant/polymer intercalations involved electronic interactions between dopant/polymer sub-lattices. The electroactive composites displayed moderately fast electrode kinetics. A composite-based biosensor for hydrogen peroxide reached a steady state current of 9.2 and 5.3 µA, and when fine-tuned for diazinon detection gave % inhibitions to be 41 and 81% for the polyaniline and poly-o-methoxyaniline biosensors, respectively.