Sensor Chip Surface Research Articles

Development of Rapid, Sensitive, and Effective Plasmonic Nanosensor for the Detection of Vitamins in Infact Formula and Milk Samples

The aim of the present study is to develop a surface plasmon resonance sensor for the detection of vitamin B2, vitamin B9, and vitamin B12 in food samples by using the molecular imprinting technique. The vitamin B2, vitamin B9, and vitamin B12 imprinted and the non-imprinted surface plasmon resonance sensor chip surfaces were characterized by using contact angle measurements, atomic force microscopy, ellipsometry, and Fourier transform infrared-attenuated total reflectance. The real-time detection of vitamin B2, vitamin B9, and vitamin B12 was analyzed by using aqueous solutions in the concentration range of 0.01 ng/mL − 10 ng/mL for vitamin B2, 0.1 ng/mL − 8.0 ng/mL for vitamin B9, and 0.01 ng/mL − 1.5 ng/mL for vitamin B12. The limit of detection values was calculated as 1.6×10−4 ng/mL for vitamin B2, 13.5×10−4 ng/mL for vitamin B9, and 2.5×10−4 ng/mL for vitamin B12, respectively. Selectivity experiments were performed by using vitamin B1 and vitamin B6. The reproducibility of surface plasmon resonance sensors was investigated both on the same day and on different days for four times. Validation studies of the prepared surface plasmon resonance (SPR) sensors were performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS).

An enzyme-free surface plasmon resonance imaging biosensing method for highly sensitive detection of microRNA based on catalytic hairpin assembly and spherical nucleic acid

MicroRNAs (miRNAs), considered as therapeutic targets and biomarkers, play important roles in biological processes. Herein, an enzyme-free surface plasmon resonance imaging (SPRi) biosensing method has been developed for miRNA detection based on catalytic hairpin assembly and spherical nucleic acid. The hairpin H1 tethered on the surface of the sensor chip is unfolded by miRNA, and then the hybridized miRNA is released through the displacement of the hairpin H2 for the successive hybridization and assembly process. The emerging DNA fragments on the sensor chip surface after hairpins assembly are further used to hybridize with spherical nucleic acid, inducing a remarkably amplified SPR signal. This biosensing method is highly sensitive to miRNA with a detection limit of 53.7 fM and a linear range of 4 orders of magnitude. Moreover, the biosensor demonstrates good specificity and has the ability to distinguish members of homologous miRNA family even with single base differences. Thus, the SPRi biosensing method may hold a great promise for further application in early clinical diagnosis.

Morphological analysis of Apolipoprotein E binding to A\u03b2 Amyloid using a combination of Surface Plasmon Resonance, Immunogold Labeling and Scanning Electron Microscopy

BackgroundImmunogold labeling in combination with transmission electron microscopy analysis is a technique frequently used to correlate high-resolution morphology studies with detailed information regarding localization of specific antigens. Although powerful, the methodology has limitations and it is frequently difficult to acquire a stringent system where unspecific low-affinity interactions are removed prior to analysis.ResultsWe here describe a combinatorial strategy where surface plasmon resonance and immunogold labeling are used followed by a direct analysis of the sensor-chip surface by scanning electron microscopy. Using this approach, we have probed the interaction between amyloid-β fibrils, associated to Alzheimer’s disease, and apolipoprotein E, a well-known ligand frequently found co-deposited to the fibrillar form of Aβ in vivo. The results display a lateral binding of ApoE along the amyloid fibrils and illustrates how the gold-beads represent a good reporter of the binding.ConclusionsThis approach exposes a technique with generic features which enables both a quantitative and a morphological evaluation of a ligand-receptor based system. The methodology mediates an advantage compared to traditional immunogold labeling since all washing steps can be monitored and where a high stringency can be maintained throughout the experiment.

Development of a Graphene-Based Surface Plasmon Resonance Optical Sensor Chip for Potential Biomedical Application.

The emergence of unintentional poisoning and uncontrolled vector diseases have contributed to sensor technologies development, leading to the more effective detection of diseases. In this study, we present the combination of graphene-based material with surface plasmon resonance technique. Two different graphene-based material sensor chips were prepared for rapid and quantitative detection of dengue virus (DENV) and cobalt ion (Co2+) as an example of typical metal ions. As the fundamental concept of surface plasmon resonance (SPR) sensor that relies on the refractive index of the sensor chip surface, this research focused on the SPR signal when the DENV and Co2+ interact with the graphene-based material sensor chip. The results demonstrated that the proposed sensor-based graphene layer was able to detect DENV and Co2+ as low as 0.1 pM and 0.1 ppm respectively. Further details in the detection and quantification of analyte were also discussed in terms of sensitivity, affinity, and selectivity of the sensor.

Rapid sensing of Cu+2 in water and biological samples by sensitive molecularly imprinted based plasmonic biosensor

In this study, copper (II) ion (Cu+2) imprinted poly(hydroxyethyl methacrylate-N-metacryloyl-(L)-cysteine methyl ester [PHEMAC-Cu+2] nanoparticles were synthesized by two-phase mini-emulsion polymerization method and applied to the SPR sensor chip surface for the selective determination of the Cu+2 ions in both aqueous solution, Cu+2-spiked artificial urine and physiological serum samples to investigate the effects of metabolite residues during the analysis. The non-imprinted [PHEMAC] nanoparticles were synthesized by applying the same procedure for the [PHEMAC-Cu+2] nanoparticle synthesis except the addition of Cu+2 ions as a control experiment to evaluate the selectivity of the [PHEMAC-Cu+2] nanoparticles. Roughness differences between [PHEMAC-Cu+2] and [PHEMAC] nanoparticles showed that the imprinting process of Cu+2 ions was performed successfully. [PHEMAC-Cu+2] and [PHEMAC] SPR biosensors prepared by attaching nanoparticles onto the surface of sensor chips, were characterized by atomic force microscope, ellipsometer, contact angle measurements.Langmuir adsorption model was found the most applicable model for this affinity system. Results showed that Cu+2 affinity regions on the surface of [PHEMAC-Cu+2] SPR biosensor were homogeneously distributed and have a monolayer structure. Having the high imprinting efficiency with the imprinting factor of 4.74, the [PHEMAC-Cu+2] SPR biosensor was found to show more selectivity towards the target Cu+2 than the non-imprinted [PHEMAC] SPR biosensor. The selectivity studies of [PHEMAC-Cu+2] SPR biosensors for Cu+2 detection were investigated by using Zn+2 and Ni+2 solutions selected as competitor molecules. The results of intraday and interday precision studies were carried out to ascertain the reproducibility of the proposed method and reported as percent relative standard deviation (%RSD) value.

A concise method for quantitative analysis of interactions between lipids and membrane proteins

Although interactions between lipids and membrane proteins (MPs) have been considered crucially important for understanding the functions of lipids, lack of useful and convincing experimental methods has hampered the analysis of the interactions. Here, we developed a surface plasmon resonance (SPR)-based concise method for quantitative analysis of lipid-MP interactions, coating the sensor chip surface with self-assembled monolayer (SAM) with C6-chain. To develop this method, we used bacteriorhodopsin (bR) as an MP, and examined its interaction with various types of lipids. The merits of using C6-SAM-modified sensor chip are as follows: (1) alkyl-chains of SAM confer a better immobilization of MPs because of the efficient preconcentration due to hydrophobic contacts; (2) SAM provides immobilized MPs with a partial membranous environment, which is important for the stabilization of MPs; and (3) a thinner C6-SAM layer (1 nm) compared with MP size forces the MP to bulge outward from the SAM surface, allowing extraneously injected lipids to be accessible to the hydrophobic transmembrane regions. Actually, the amount of bR immobilized on C6-SAM is 10 times higher than that on a hydrophilic CM5 sensor chip, and AFM observations confirmed that bR molecules are exposed on the SAM surface. Of the lipids tested, S-TGA-1, a halobacterium-derived glycolipid, had the highest specificity to bR with a nanomolar dissociation constant. This is consistent with the reported co-crystal structure that indicates the formation of several intermolecular hydrogen bonds. Therefore, we not only reproduced the specific lipid-bR recognition, but also succeeded in its quantitative evaluation, demonstrating the validity and utility of this method.

Interferometric detection of chloramphenicol via its immunochemical recognition at polymer-coated nano-corrugated surfaces

Three types of polymer-coated glass slides were prepared by acylation of their γ–aminopropyl derivatives with succinic anhydride or poly(p-nitrophenyl acylate) (PNPA) at various reaction times. The variations in duration of chemical adsorption of PNPA allowed control over nano-corrugation of the reactive surfaces employed for biosensing. The visually smooth and optically clear polymer-coated slides were used for immobilization of a conjugate of bovine serum albumin with chloramphenicol (CAP) and label-free measurement of concentration of this antibiotic. The sensor chips prepared in this way provided strong biosensing response due to fast biomolecular interactions and efficient competition of the target analyte during indirect immunoassay. The kinetics of formation and dissociation of immune complexes “conjugate-antibody” on the sensor chip surface was registered by the label-free methods of spectral-correlation and spectral-phase interferometry. The average thickness of immune complex adsorption layers was in the range of 5–12 nm. The highest value of kinetic association constant (kon = 5 × 104 M−1s−1) and the lowest equilibrium dissociation constant (Kdiss = 2 × 10-8 M−1) were achieved with the chips originated in the PNPA-coated slides, which were produced under short (2 min) polymer chemisorption and exhibited nano-corrugation almost unchanged with respect to the parent glass, according to the atomic-force microscopy. The detection limit for CAP detection was 80 pg/mL, and the dynamic range was 5 orders. The proposed methods for fabrication of sensor chips with nano-corrugated surface, the results of their characterization, as well as the developed assay can be employed for high-throughput recording of multiple molecular interactions by spectral interferometry and in a wide range of biosensing platforms for registration of biologically active compounds in medicine, veterinary, food control and ecological monitoring.

Affinity profiling of monoclonal antibody and antibody-drug-conjugate preparations by coupled liquid chromatography-surface plasmon resonance biosensing

Monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) are highly potent biopharmaceuticals designed for targeted cancer therapies. mAbs and ADCs can undergo modifications during production and storage which may affect binding to target receptors, potentially altering drug efficacy. In this work, liquid chromatography was coupled online to surface plasmon resonance (LC-SPR) to allow label-free affinity evaluation of mAb and ADC sample constituents (size and charge variants), under near-native conditions. Trastuzumab and its ADC trastuzumab emtansine (T-DM1) were used as a test sample and were analyzed by aqueous size-exclusion chromatography (SEC)-SPR before and after exposure to aggregate-inducing conditions. SEC-SPR allowed separation of the formed aggregates and measurement of their affinity towards the ligand-binding domain of the human epidermal growth factor receptor 2 (HER2) receptor immobilized on the surface of the SPR sensor chip. The monomer and aggregates of the mAb and ADC were shown to have similar antigen affinity. Conjugation of drugs to trastuzumab appeared to accelerate the aggregate formation. In addition, cation-exchange chromatography (CEX) was coupled to SPR enabling monitoring the maximum ligand-analyte binding capacity (Rmax) of individual charge variants present in mAbs. Deamidated species and lysine variants in trastuzumab sample were separated but did not show different binding affinities to the immobilized HER2-binding domain. In order to allow protein variant assignment, parallel MS detection was added to the LC-SPR setup using a column effluent split. The feasibility of the LC-MS/SPR system was demonstrated by analysis of trastuzumab and T-DM1 providing information on antibody glycoforms and/or determination of the drug-to-antibody ratio (DAR), while simultaneously monitoring binding of eluting species to HER2.Graphical abstractᅟ

Design and analysis of a three-dimensional millimeter-wave frequency-shift based CMOS biosensor using vertically stacked spiral inductors in LC oscillators

This paper proposes a novel millimeter-wave frequency-shift based CMOS biosensor capable of providing three-dimensional (3D) resolution. The vertical resolution from the sensor chip surface can be obtained by using vertically stacked on-chip inductors of LC oscillators in dual metal layers, which enables 3D detection of biomolecular target. The LC oscillators with spiral inductors stacked in upper and lower metal layers produce different shifts in the frequencies from the desired resonant frequency of 60 GHz due to frequency dependent complex relative permittivity of targeted bio-molecular structure. The vertical resolution of the sensor can also be improved by changing the supply voltage. The effectiveness of the proposed approach is verified by fabricating a test chip in 65 nm CMOS process. The measurement results show variation in the resonant frequency shifts due to upper and lower spiral inductors of LC oscillators which demonstrates the capability of the proposed biosensor to provide 3D resolution.

Overcoming non-specific binding to measure the active concentration and kinetics of serum anti-HLA antibodies by surface plasmon resonance

Human leukocyte antigen (HLA) donor-specific antibodies are key serum biomarkers for assessing the outcome of transplanted patients. Measuring their active concentration, i.e. the fraction that really interacts with donor HLA, and their affinity could help deciphering their pathogenicity. Surface plasmon resonance (SPR) is recognized as the gold-standard for measuring binding kinetics but also active concentrations, without calibration curves. SPR-based biosensors often suffer from non-specific binding (NSB) occurring with the sensor chip surface and the immobilized targets, especially for complex media such as human serum. In this work we show that several serum treatments such as dialysis or IgG purification reduce NSB but insufficiently for SPR applications. We then demonstrate that the NSB contribution to the SPR signal can be eliminated to determine precisely and reliably the active concentration and the affinity of anti-HLA antibodies from patients' sera. This was achieved even at concentrations close to the limit of quantification of the method, in the 0.5–1 nM range. The robustness of the assay was demonstrated by using a wide range of artificially generated NSB and by varying the density of the targets captured onto the surface. The assay is of general interest and can be used with molecules generating strong NSB, as far as a non-cognate target structurally close to the target can be captured on the same flow cell, in a different binding cycle. Compared with current fluorescence-based methods that are semi-quantitative, we expect this SPR-based assay to help better understanding anti-HLA antibodies pathogenicity and improving organ recipients’ management.

Carbodiimide-mediated immobilization of acidic biomolecules on reversed-charge zwitterionic sensor chip surfaces.

The carbodiimide-mediated amine coupling of protein ligands to sensor chips coated with anionic polycarboxylate hydrogels, such as carboxymethyl dextran, is the predominant covalent immobilization procedure utilized in optical biosensors, namely surface plasmon resonance (SPR) biosensors. Usually, electrostatic interactions at a slightly acidic pH and low ionic strength are employed to efficiently accumulate neutral and basic ligands on the chip surface, which are then covalently coupled by surface-bound active N-hydroxysuccinimide (NHS) esters. Unfortunately, this approach is not suitable for acidic proteins or other ligands with low isoelectric points (IEPs), such as nucleic acids, because the charge density of the polycarboxylates is greatly reduced at acidic pH or because electrostatic attraction cannot be achieved. To overcome these drawbacks, we have established a charge-reversal approach that allows the preconcentration of acidic proteins above their IEPs. A precisely controlled amount of tertiary amines is applied to reverse the previous anionic surface charge while maintaining carbodiimide compatibility with future protein immobilization. The mechanism of this reversed-charge immobilization approach was demonstrated employing protein A as a model protein and using attenuated total reflectance Fourier transform infrared spectroscopy, dynamic contact angle measurements, colorimetric quantification, and SPR analysis to characterize surface derivatization. Furthermore, even though it had previously proven impossible to preconcentrate DNA electrostatically and to covalently couple it to polyanionic chip surfaces, we demonstrated that our approach allowed DNA to be preconcentrated and immobilized in good yields. Graphical abstract Principle of the covalent immobilization of acidic ligands on reversed-charge zwitterionic sensor chip surfaces.

A direct determination of AFBs in vinegar by aptamer-based surface plasmon resonance biosensor

Aflatoxin (AFB) is one of the most toxic fungal metabolites produced by Aspergillus flavus, which may contaminate food and agricultural products. Herein, an aptamer-based surface plasmon resonance (SPR) biosensor was developed to detect AFBs. The chosen aptamer showed comparable interaction with the two AFBs, namely aflatoxin B1 (AFB1) and aflatoxin B2 (AFB2). Such phenomenon was rarely reported, and might lead to a simultaneous detection of both AFBs. In this study, AFB1 was used to systematically establish the detection method. In the SPR system, streptavidin proteins were immobilized on the surface of a CM5 sensor chip as a cross-linker and biotin-aptamers were captured through streptavidin-biotin interaction. After optimization, the assay showed a dynamic range between 0.09 and 200 ng mL−1 (linear range from 1.5 to 50 ng mL−1 and a LOD of 0.19 ng mL−1) of AFB1 in buffer. As expected, the aptasensor showed high specificity towards AFB1 and AFB2, but hardly bound to other toxins with similar structures, including ochratoxin A (OTA), ochratoxin B (OTB), Zeralenone (ZEA) and T-2 toxin (T-2). Determination of AFB1 in vinegar was further performed using the SPR biosensor. Recoveries of AFB1 from spiked samples ranged from 96.3 to 117.8%. The developed SPR assay is a simple, fast and sensitive approach for the detection of residual AFBs in agricultural products and foodstuffs like vinegar.

QCM Biosensor Based on Polydopamine Surface for Real-Time Analysis of the Binding Kinetics of Protein-Protein Interactions.

A quartz crystal microbalance (QCM) biosensor based on polydopamine (PDA) surface was developed for real-time analysis of the binding kinetics of protein-protein interactions. The biosensor was fabricated by simply immersing the gold sensor chip into an aqueous dopamine solution at pH 8.5 leading to a spontaneous deposition of PDA film onto the sensor chip surface, which was followed by incubation with the protein to immobilize it onto the PDA-coated sensor chip surface via Michael addition and/or Schiff base reactions. In this paper, the interaction between monoclonal anti-myoglobin 7005 antibody (IgG1) and its antigen human cardiac myoglobin was used as a model system for real-time analysis of biomolecule interactions on the biosensor surface. The kinetic parameters of the interaction between anti-myoglobin 7005 and myoglobin were studied on the biosensor surface, which were consistent with the results obtained via amine coupling. The biosensor based on PDA surface has excellent regenerability, reproducibility, and specificity. Compared with the most frequently/typically used amine coupling method for immobilization of proteins on carboxylated substrates, the modification methodology presented in this paper is simple, mild and is not subjected to the limitations of the isoelectric point (pI) of the protein. In addition, the PDA biosensor chip can be easily reused, which makes QCM biosensor analysis more efficient and cost effective.

Rational Design of Peptide-Functionalized Surface Plasmon Resonance Sensor for Specific Detection of TNT Explosive

In this study, a rationally-designed 2,4,6-trinitrotoluene (TNT) binding peptide derived from an amino acid sequence of the complementarity-determining region (CDR) of an anti-TNT monoclonal antibody was used for TNT detection based on a maleimide-functionalized surface plasmon resonance (SPR) sensor. By antigen-docking simulation and screening, the TNT binding candidate peptides were obtained as TNTHCDR1 derived from the heavy chain of CDR1, TNTHCDR2 derived from CDR2, and TNTHCDR3 from CDR3 of an anti-TNT antibody. The binding events between candidate peptides and TNT were evaluated using the SPR sensor by direct determination based on the 3-aminopropyltriethoxysilane (APTES) surface. The TNT binding peptide was directly immobilized on the maleimide-functionalized sensor chip surface from N-γ-maleimidobutyryl-oxysuccinimide ester (GMBS). The results demonstrated that peptide TNTHCDR3 was identified and selected as a TNT binding peptide among the other two candidate peptides. Five kinds of TNT analogues were also investigated to testify the selectivity of TNT binding peptide TNTHCDR3. Furthermore, the results indicated that the APTES-GMBS-based SPR sensor chip procedure featured a great potential application for the direct detection of TNT.

Optimization of 3D Surfaces of Dextran with Different Molecule Weights for Real-Time Detection of Biomolecular Interactions by a QCM Biosensor.

Quartz crystal microbalance (QCM) has been extensively applied in real-time and label-free biomolecular interaction studies. However, the sensitive detection by QCM technology remains challenging, mainly due to the limited surface immobilization capacity. Here, a three-dimensional (3D) carboxymethyl dextran coated gold sensor chip surface was successfully fabricated with dextran of different molecular weight (100, 500 and 2000 kDa, respectively). To evaluate the 3D carboxymethyl dextran surface immobilization capacity, the 3D surface was used for studying antigen–antibody interactions on the QCM biosensor. The results showed that the protein immobilization capacity of the 3D carboxymethyl dextran (2000 kDa) surface exceeded more than 4 times the capacity of the 2D carboxyl surface, and 2 times the capacity of the traditional 3D carboxymethyl dextran (500 kDa) surface. Furthermore, the kinetic and affinity properties of antigen–antibody interactions were performed. Most notably, the optimized 3D carboxymethyl dextran (2000 kDa) surface could be used for small molecule detection, where the binding of biotinylated oligo (0.67 kDa) reached 8.1 Hz. The results confirmed that a 3D carboxymethyl dextran (2000 kDa) surface can be exploited for sensitive detection of low molecular weight analytes, which have great potential applications for characterizing the interactions between small molecule drugs and proteins.

Magnetic nanoparticle enhanced surface plasmon resonance sensor for estradiol analysis

An enhanced surface plasmon resonance (SPR) biosensor system using magnetic nanoparticles as amplifying element was developed for the real-time determination of estradiol (E2) based on indirect competition method. The anti-estradiol monoclonal antibody (E2-mAb) was conjugated with the magnetic nanoparticles (MNPs) via protein A for signal amplification to improve the detection sensitivity. The chitosan, spin-coated on the sensor chip surface, was used to immobilize the antigen (E2-BSA) by glutaraldehyde. The chitosan could significantly improve the performance and stability of the immobilized E2-BSA. During the detection, E2-BSA competed with E2 in samples for binding with the E2-mAb-MNPs conjugates. Then the SPR response decreased in the presence of E2 because E2 prevented the combination of E2-mAb-MNPs conjugates and E2-BSA. In other words, the response of SPR sensor was inversely proportional to E2 concentration. Assay parameters, such as the amount of antibody and MNPs, the amount of immobilized E2-BSA and E2-mAb-MNPs concentration, were optimized in detail and E2-spiked milk samples were detected. A good linear relationship was obtained between inhibition and lgC(E2) ranging from 1.95 to 2000ng/mL and the limit of detection (LOD) was 0.81ng/mL. The result implied that the detection sensitivity was improved compared with the traditionnal sensor without magnetic nanoparticles. Meanwhile, an ELISA method was conducted to detect E2 in milk as comparation with the enhanced SPR method, and the results indicated that these two methods had good consistency. The present study demonstrated a presumable general way and the enhanced sensors possessed a promising application for detection of various kinds of small molecules.

Transport of melamine by a new generation of nano-material membranes containing carbon nanotubes and determination with surface plasmon resonance

This study developed a surface plasmon resonance process for the determination of melamine by using a carbon nanotube inclusion polymeric membrane for the first time in the literature. It was prepared with a low-cost biosensor, which allowed the determination of melamine by molecular imprinted polymeric films in a one-step, selective, and sensitive way. For this purpose, the molecular imprinted sensors intended for the determination of melamine from a milk sample were developed. The study was achieved in three stages. In the first stage, melamine was transported by a new generation of carbon nanotube membrane, which was improved by adding nanomaterials. In the second stage, printed biosensor chips were produced for the determination of melamine. In the last stage, prepared biosensor chips were used for the determination and kinetic analysis of melamine in a surface plasmon resonance system. Melamine was transported from the milk sample with 82.87% efficiency at the end of ten days. The determination of melamine was achieved in a highly sensitive, selective, and easy way by preparing a melamine printed polymer nanofilm layer on the surface of the SPR's sensor chip. By using new generation carbon nanotube membranes (CNT/PIM), nano-reinforced membranes were developed, which included high permeability and mechanical strength against contamination, and the fulfill an active layer of the membrane was minimized.

Evaluation of the Interaction between Pesticides and a Cell Membrane Model by Surface Plasmon Resonance Spectroscopy Analysis.

A surface plasmon resonance (SPR) spectroscopy analysis was used for the characterization of the interaction between pesticides and a cell membrane model. A liposome was immobilized onto the surface of the SPR sensor chip (L1), and the lipid bilayer membrane formed on the sensor chip was regarded as the cell membrane model. The solution containing a pesticide was flowed onto the sensor chip, and an SPR sensorgram, which reflected the interaction between the pesticide and the lipid bilayer membrane, was obtained. As the results, the pattern and strength of the interaction of the pesticides with the cell membrane model were visualized and quantified. Triflumizole, hexythiazox, and pentachlorophenol showed a strong interaction with the lipid bilayer. It is well-known that triflumizole and pentachlorophenol interact with the membrane and reveal toxicities for cell membranes. In addition, there was a tendency for higher residual ratios to be observed when the no observable adverse effect level (NOAEL) values for chronic toxicity (1 year toxicity study in dogs) were lower. We suggest that a novel parameter for the evaluation or presumption of the behaviors and chronic toxicities of pesticides is obtained by the presented method.

Differential imaging of the metabolism of bacteria and eukaryotic cells based on light-addressable potentiometric sensors

A light-addressable potentiometric sensor (LAPS) is a field-effect-based potentiometric sensor with an electrolyte/insulator/semiconductor (EIS) structure, which is able to monitor analyte concentrations of (bio-)chemical species in aqueous solutions in a spatially resolved way. Therefore, it is also an appropriate tool to record 2D-chemical images of concentration variations on the sensor surface. In the present work, two differential, LAPS-based measurement principles are introduced to determine the metabolic activity of Escherichia coli (E. coli) K12 and Chinese hamster ovary (CHO) cells as test microorganisms. Hereby, we focus on i) the determination of the extracellular acidification rate (ΔpH/min) after adding glucose solutions to the cell suspensions; and ii) recording the amplitude increase of the photocurrent (Iph) related to the produced acids from E. coli K12 bacteria and CHO cells on the sensor surface by 2D-chemical imaging. For this purpose, 3D-printed multi-chamber structures were developed and mounted on the planar sensor-chip surface to define four independent compartments, enabling differential measurements with varying cell concentrations. The differential concept allows eliminating unwanted drift effects and, with the four-chamber structures, measurements on the different cell concentrations were performed simultaneously, thus reducing also the overall measuring time.

Aptamer based surface plasmon resonance sensor for aflatoxin B1

The authors describe a surface plasmon resonance (SPR) based aptasensor for the carcinogenic mycotoxin aflatoxin B1 (AFB1) in a direct assay format. The aptamer is immobilized on the surface of a commercial sensor chip, and the SPR signal increases on binding of AFB1. The sensor chip can be fully regenerated by passing a flow of buffer over it upon which bound AFB1 dissociates from the aptamer. The biosensor works in the 0.4 nM to 200 nM AFB1 concentration range and has a 0.4 nM detection limit. It allows AFB1 to be determined in complex samples such as diluted red wine and beer. The assay is sensitive, and the chip is easily regenerated and stable. The method therefore overcomes certain limitations of antibody-based SPR assays and of competitive SPR assays for AFB1.