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An 8-pixel micromachined quartz crystal resonator array with a fundamental resonance frequency of 66 MHz has been designed, fabricated, and tested. A compact impedance-spectrum-analyzer electronic interface has been developed and combined with the quartz resonator array to form the biosensing system. The sensor array was calibrated using water-glycerol solutions, and the performance was found to be exactly as expected. Measurement of the crosstalk between the sensor pixels showed an isolation of ~30 dB. Selective functionalization of the pixels was achieved through the use of aqueous 3, 3'-Dithiobis (sulfosuccinimidylpropionate) (DTSSP) molecules. The adsorption of avidin on DTSSP gave a frequency signal of 60 kHz in comparison to unfunctionalized pixels. The specific adsorption of avidin on functionalized pixeis was confirmed through fluorescence microscopy. Comparing the performance of the micromachined quartz crystal microbalance (QCM) with a commercial 5-MHz device, we found that the micromachined QCM has a 4.25 times higher signal-to-noise ratio. Based on the measurement of the noise and using three times the frequency noise as the limit for the detection of avidin molecules, we expect to resolve a minimum of ~1/960 of a monolayer of avidin corresponding to an aerial mass density resolution of 0.7 ng/[cm.sup.2]. [2008-0196] Index Terms--Biosensor array, impedance spectrum analyzer, quartz crystal microbalance (QCM), quartz micromachining.

Transducers for high-temperature applications often use buffer rods. The influence of noise sources as a function of geometry and operating environment is investigated and reported.

The question of whether an oscillator-based measurement is adequate in a liquid environment is answered in the affirmative.

Viscoelastic contributions to the frequency shift in the sensor response generated from viscoelastic coatings were examined.

Data from quartz crystal resonators can, with the proposed method, swiftly produce shear moduli of polymer films. Electrical admittance of the quartz crystal can produce acoustic load impedance, then a family of approximations is applied, and the approximation is improved through an iterative procedure.

The sensitivity of acoustic-wave microsensors coated with a viscoelastic film to mass changes and film modulus (changes) is examined. The study analyzes the acoustic load at the interface between the acoustic device and the coating. The acoustic load carries information about surface mass and film modulus; its determination has no restrictions in film thickness. Two regimes of film behavior can be distinguished: the gravimetric regime, where the sensor response is mainly mass sensitive, and the nongravimetric regime, where viscoelasticity gains influence on the sensor response. We develop a method, which allows the assignment of the sensor signal to a gravimetric or a nongravimetric response. The critical value can be determined from oscillator measurements. The related limits for the coating thickness are not the same for the coating procedure and mass accumulation during chemical sensing. As an example, we present results from a 10 MHz quartz crystal resonator.

The rough surface of a quartz resonator vibrating in thickness-shear model (TSM) traps liquid which act as a mass effect to the crystal. The mass effect relies on both the liquid mass enclosed in the surface cavities and the liquid properties and crystal surface features. The trapping factor is introduced to analyze the way the liquid mass effect operates.

Concentration gradients that occur in large industrial‐scale bioreactors due to mass transfer limitations have significant effects on process efficiency. Hence, it is desirable to investigate the response of strains to such heterogeneities to reduce the risk of failure during process scale‐up. Although there are various scale‐down techniques to study these effects, scale‐down strategies are rarely applied in the early developmental phases of a bioprocess, as they have not yet been implemented on small‐scale parallel cultivation devices. In this study, we combine mechanistic growth models with a parallel mini‐bioreactor system to create a high‐throughput platform for studying the response of Escherichia coli strains to concentration gradients. As a scaled‐down approach, a model‐based glucose pulse feeding scheme is applied and compared with a continuous feed profile to study the influence of glucose and dissolved oxygen gradients on both cell physiology and incorporation of noncanonical amino acids into recombinant proinsulin. The results show a significant increase in the incorporation of the noncanonical amino acid norvaline in the soluble intracellular extract and in the recombinant product in cultures with glucose/oxygen oscillations. Interestingly, the amount of norvaline depends on the pulse frequency and is negligible with continuous feeding, confirming observations from large‐scale cultivations. Most importantly, the results also show that a larger number of the model parameters are significantly affected by the scale‐down scheme, compared with the reference cultivations. In this example, it was possible to describe the effects of oscillations in a single parallel experiment. The platform offers the opportunity to combine strain screening with scale‐down studies to select the most robust strains for bioprocess scale‐up. [ABSTRACT FROM AUTHOR]

N-Glycosylation in the endoplasmic reticulum is an essential protein modification and highly conserved in evolution from yeast to humans. The key step of this pathway is the transfer of the lipid-linked core oligosaccharide to the nascent polypeptide chain, catalyzed by the oligosaccharyltransferase complex. Temperature-sensitive oligosaccharyltransferase mutants of Saccharomyces cerevisiae at the restrictive temperature, such as wbp1-1, as well as wild-type cells in the presence of the N-glycosylation inhibitor tunicamycin display typical apoptotic phenotypes like nuclear condensation, DNA fragmentation, phosphatidylserine translocation, caspase-like activity, and reactive oxygen species accumulation. Since deletion of the yeast metacaspase YCA1 did not abrogate this death pathway, we postulated a different proteolytic process to be responsible. Here, we show that Kex1 protease is involved in the programmed cell death caused by defective N-glycosylation. Its disruption decreases caspase-like activity, production of reactive oxygen species, and fragmentation of mitoehondria and, conversely, improves growth and survival of cells. Moreover, we demonstrate that Kex1 contributes also to the active cell death program induced by acetic acid stress or during chronological aging, suggesting that Kex1 plays a more general role in cellular suicide of yeast. [ABSTRACT FROM AUTHOR]

For decades impedance spectroscopy is used in technical laboratories and research departments to investigate effects or material characteristics that affect the impedance spectrum of the sensor. Establishing this analytical approach for process automation and stand-alone applications will deliver additional and valuable information beside traditional measurement techniques such as the measurement of temperature, flow rate, and conductivity, among others. As yet, most of the current impedance analysis methods are suited for laboratory applications only since they involve stand-alone network analyzers that are slow, expensive, large, or immobile. Furthermore, those systems offer a large range of functionality that is not being used in process control and other fields of application. We developed a sensor interface based on high speed direct digital signal processing offering wideband impedance spectrum analysis with high resolution for frequency adjustment, excellent noise rejection, very high measurement rate, and convenient data exchange to common interfaces. The electronics has been implemented on two small circuit boards and it is well suited for process control applications such as monitoring phase transitions, characterization of fluidal systems, and control of biological processes. The impedance spectrum analyzer can be customized easily for different measurement applications by adapting the appropriate sensor module. It has been tested for industrial applications, e.g., dielectric spectroscopy and high temperature gas analysis. [ABSTRACT FROM AUTHOR]

Acoustic-wave-based sensors are commonly known as mass-sensitive devices. However, acoustic chemical and biochemical sensors also face so-called non-gravimetric effects, especially if they work in a liquid environment. The one-dimensional transmission-line model (TLM) is a powerful tool, which considers the influence of geometric and material properties on the sensor transduction mechanism, most importantly the influence of viscoelastic phenomena. This paper demonstrates the concept of modelling acoustic microsensors on quartz crystal resonators. Particular attention is paid to special cases which allow for simplifications or specific solutions of the TLM, like the acoustic load concept (ALC), the BVD model or the Sauerbrey equation. Deviations from the one-dimensional assumption of the TLM are suspected to significantly contribute to the acoustic sensor response in biosystems. We therefore introduce a generalization of the ALC to get access to two- or three-dimensional effects, which are up to now not considered in the TLM. As examples, signatures of interfacial phenomena or non-uniform films are discussed. [ABSTRACT FROM AUTHOR]

The application of resonant sensors such as quartz crystal microbalance (QCM) resonators requires interface electronics to measure parameters that characterize sufficiently the resonant behavior of the sensor due to effects under investigation. Common oscillators as sensor electronics have two major disadvantages. Since they measure the resonant frequency shift as the only parameter their employment is restricted to mass sensing of thin and rigid films in terms of QCM. Because of spurious phase shifts the measured resonant frequency may be erratic. In order to characterize the sensor itself or a material on the sensor surface more parameters such as damping are to be measured. Therefore, a sensor electronics was developed that precisely acquires the impedance spectrum of the resonant sensor in a small frequency range which reflects properties like thickness of a sensor coating, its density, and shear moduli or the density-viscosity product when measuring in liquids. Originally developed as a sensor interface electronics this novel device can help researchers to investigate any effects or material parameters affecting the impedance spectrum of the sensor. In this article the measurement concept of the network analysis based device will be explained followed by details of realization on single printed cardboard. Measurement errors are estimated by simulation and by a comparison of measurement results from a commercially available network analyzer as a reference. Examples of laboratory and industrial applications will be given. © 2001 American Institute of Physics. [ABSTRACT FROM AUTHOR]

Presents information on a study which analyzed ultrasonic density sensors for liquids. Requirements of densitometers and mass flow meters for process automation; Principles of ultrasonic measurement and error analysis; Description of the experimental setup; Conclusions.

Insufficient mixing in large-scale bioreactors provokes gradient zones of substrate, dissolved oxygen (DO), pH, and other parameters. E. coli responds to a high glucose, low oxygen feeding zone with the accumulation of mixed acid fermentation products, especially formate, but also with the synthesis of non-canonical amino acids, such as norvaline, norleucine and β-methylnorleucine. These amino acids can be mis-incorporated into recombinant products, which causes a problem for pharmaceutical production whose solution is not trivial. While these effects can also be observed in scale down bioreactor systems, these are challenging to operate. Especially the high-throughput screening of clone libraries is not easy, as fed-batch cultivations would need to be controlled via repeated glucose pulses with simultaneous oxygen limitation, as has been demonstrated in well controlled robotic systems. Here we show that not only glucose pulses in combination with oxygen limitation can provoke the synthesis of these non-canonical branched-chain amino acids (ncBCAA), but also that pyruvate pulses produce the same effect. Therefore, we combined the enzyme-based glucose delivery method Enbase® in a PALL24 mini-bioreactor system and combined repeated pyruvate pulses with simultaneous reduction of the aeration rate. These cultivation conditions produced an increase in the non-canonical branched chain amino acids norvaline and norleucine in both the intracellular soluble protein and inclusion body fractions with mini-proinsulin as an example product, and this effect was verified in a 15 L stirred tank bioreactor (STR). To our opinion this cultivation strategy is easy to apply for the screening of strain libraries under standard laboratory conditions if no complex robotic and well controlled parallel cultivation devices are available. [ABSTRACT FROM AUTHOR]

Studies the interface circuits of quartz-crystal-microbalance sensors in liquids. Fundamentals of the measuring principle; Advantages and drawbacks of suitable interface circuits; Requirements of oscillators as interface circuits.

States that a transmission line equivalent circuit for piezoeletric transducers has been modified to provide modelling of lossy piezoceramic transducers. Model of the piezoelectic thickness-mode transducer; SPICE implementation; Examples of calculations done to demonstrate the application of the model; Uses of low-Q piezoceramic transducer; Conclusions of the study.

Downstream processing in the manufacturing biopharmaceutical industry is a multistep process separating the desired product from process‐ and product‐related impurities. However, removing product‐related impurities, such as product variants, without compromising the product yield or prolonging the process time due to extensive quality control analytics, remains a major challenge. Here, we show how mechanistic model‐based monitoring, based on analytical quality control data, can predict product variants by modeling their chromatographic separation during product polishing with reversed phase chromatography. The system was described by a kinetic dispersive model with a modified Langmuir isotherm. Solely quality control analytical data on product and product variant concentrations were used to calibrate the model. This model‐based monitoring approach was developed for an insulin purification process. Industrial materials were used in the separation of insulin and two insulin variants, one eluting at the product peak front and one eluting at the product peak tail. The model, fitted to analytical data, used one component to simulate each protein, or two components when a peak displayed a shoulder. This monitoring approach allowed the prediction of the elution patterns of insulin and both insulin variants. The results indicate the potential of using model‐based monitoring in downstream polishing at industrial scale to take pooling decisions. [ABSTRACT FROM AUTHOR]

The capability of intelligent sensors to have more intelligence built into them continues to drive their application in areas including automotive, aerospace and defense, industrial, intelligent house and wear, medical and homeland security. In principle it is difficult to overestimate the importance of intelligent (micro) sensors or sensor systems within advanced societies but one characteristic feature is the global market for sensors, which is now about $20 billion annually. Therefore sensors or sensor systems play a dominant role in many fields from the macro sensor in manufacturing industry down to the miniaturized sensor for medical applications. The diversity of sensors precludes a complete description of the state-of-the-art; selected examples will illustrate the current situation. MEMS (microelectromechanical systems) devices are of special interest in the context of micro sensor systems. In past the main requirements of a sensor were in terms of metrological performance. The electrical (or optical) signal produced by the sensor needed to match the measure relatively accurately. Such basic functionality is no longer sufficient. Data processing near the sensor, the extraction of more information than just the direct sensor information by signal analysis, system aspects and multi-sensor information are the new demands. A shifting can be observed away from aiming to design perfect single-function transducers and towards the utilization of system-based sensors as system components. In the ideal case such systems contain sensors, actuators and electronics. They can be realized in monolithic, hybrid or discrete form—which kind is used depends on the application. In this article the state-of-the-art of intelligent sensors or sensor systems is reviewed using selected examples. Future trends are deduced. [ABSTRACT FROM AUTHOR]