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  • 101. van Noort, D.
    et al.
    Mandenius, Carl-Fredrik
    Linköpings universitet, Tekniska högskolan. Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi.
    Porous gold surfaces for biosensor applications2000Inngår i: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 15, nr 3-4, s. 203-209Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    The sensitivity of optical biosensors where the detection takes place on a planar gold surface can be improved by making the surface porous. The porosity allows a larger number of ligands per surface area resulting in larger optical shifts when interacting with specifically binding analyte molecules. The porous gold was deposited as a thin layer on a planar gold surface by electrochemical deposition in a solution of tetrachloroaurate and lead acetate. A protein, streptavidin, was adsorbed into the formed porous layer and the time course of the adsorption was monitored by in-situ ellipsometry. When the porous layer was 500 nm in thickness a six-fold increase of the ellipsometric response was obtained compared with a planar gold surface. The dependency of porosity and layer thickness was explained with a mathematical model of the gold/porous gold/protein/solution system. Copyright (C) 2000 Elsevier Science S.A.The sensitivity of optical biosensors where the detection takes place on a planar gold surface can be improved by making the surface porous. The porosity allows a larger number of ligands per surface area resulting in larger optical shifts when interacting with specifically binding analyte molecules. The porous gold was deposited as a thin layer on a planar gold surface by electrochemical deposition in a solution of tetrachloroaurate and lead acetate. A protein, streptavidin, was adsorbed into the formed porous layer and the time course of the adsorption was monitored by in-situ ellipsometry. When the porous layer was 500 nm in thickness a six-fold increase of the ellipsometric response was obtained compared with a planar gold surface. The dependency of porosity and layer thickness was explained with a mathematical model of the gold/porous gold/protein/solution system.

  • 102. van Noort, D
    et al.
    Rani, R
    Mandenius, Carl-Fredrik
    Linköpings universitet, Tekniska högskolan. Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi.
    Improving the sensitivity of a quartz crystal microbalance for biosensing by using porous gold2001Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    A quartz crystal microbalance can be used as a biosensor if biological receptor molecules (ligands) are attached to the crystal. To improve the sensitivity, the surface area of the gold electrodes on the crystal was increased by rendering the electrodes porous. This increased the response up to a factor of 3. A protein model system with human anti-myoglobin as ligand and sheep skeletal myoglobin at different concentrations as analyte was used. The quartz crystal was mounted in a flow-cell, where immobilisation, binding and regeneration of the surface were carried out. The kinetics of the model system was monitored under various experimental conditions.

  • 103.
    van Noort, Danny
    et al.
    Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi. Linköpings universitet, Tekniska högskolan.
    Rumberg, Jens
    School of Chemical and Physical Sciences, Victoria University, Wellington, New Zealand.
    Jager, Edwin
    Linköpings universitet, Institutionen för fysik, kemi och biologi, Biomolekylär och Organisk Elektronik. Linköpings universitet, Tekniska högskolan.
    Mandenius, Carl-Fredrik
    Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi. Linköpings universitet, Tekniska högskolan.
    Silicon based affinity biochips viewed with imaging ellipsometry2000Inngår i: Measurement science and technology, ISSN 0957-0233, E-ISSN 1361-6501, Vol. 11, nr 6, s. 801-Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    In this paper we report on the fabrication of an affinity biochip with a matrix of 900 targets for detection with imaging ellipsometry. Two methods of fabrication of chips are shown: one based on wet etching of a silicon surface and the other on the preparation of so-called tension wells on the silicon surface. The dispensing of reagents and ligands was performed using a pipetting robot equipped with a micro-capillary, a syringe pump and micro-stepping motors. Measurements were performed on the chips in real time with carbohydrate model substances selected for six common lectins. Affinity binding was shown for three of the tested model substances.

  • 104. Vostiar, I.
    et al.
    Tkac, J.
    Mandenius, Carl-Fredrik
    Linköpings universitet, Tekniska högskolan. Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi.
    Intracellular monitoring of superoxide dismutase expression in an Escherichia coli fed-batch cultivation using on-line disruption with at-line surface plasmon resonance detection2005Inngår i: Analytical Biochemistry, ISSN 0003-2697, E-ISSN 1096-0309, Vol. 342, nr 1, s. 152-159Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    An on-line cell disruption system for at-line monitoring of the intracellular concentration of recombinant human superoxide dismutase (rhSOD) in a genetically modified Escherichia coli strain, HMS174(DE3) (pET11a/rhSOD), in bioreactor cultivations is described. The sampled bacteria were disrupted on-line by rapid mixing with a nonionic detergent. The recombinant protein content of the lysed bacterial sample was quantitated by a subsequent surface plasmon resonance biosensor with a specific monoclonal antibody. Extraction efficiency of the monitoring system was optimized with respect to the flow rate ratio of the cell suspension and the detergent at relevant cell densities with the aim to attain rapid monitoring. Monitoring was demonstrated for a shake flask culture and a glucose-limited fed-batch cultivation. The results are compared with a traditional enzyme-linked immunosorbent assay method showing a correlation coefficient of R2 = 0.97. Extraction efficiency of rhSOD reached 95-99% at a total processing time of 1.8-2.6 min and a contact time of 0.8-1.4 min. The possibility of extending the monitoring system to other intracellular proteins is discussed. © 2005 Elsevier Inc. All rights reserved.

  • 105. Vostiar, I.
    et al.
    Tkac, J.
    Mandenius, Carl-Fredrik
    Linköpings universitet, Tekniska högskolan. Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi.
    Monitoring of the heat-shock response in Escherichia coli using an optical biosensor2003Inngår i: Analytical Biochemistry, ISSN 0003-2697, E-ISSN 1096-0309, Vol. 322, nr 2, s. 156-163Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    A surface plasmon resonance (SPR) method for monitoring the concentration of the chaperone DnaK and its relation to physiological stress response in a recombinant Escherichia coli strain subjected to heat shock is described. The DnaK protein, an abundantly occurring representative of the heat-shock proteins, was used as a marker of physiological stress. The SPR biosensor instrument was used for label-free immunoaffinity detection directly in cell culture lysates using an anti-DnaK monoclonal IgG antibody immobilized on the sensor surface. The SPR method provides a fast response (<8min) and a reproducible (RSD<2%), accurate (comparison to the direct enzyme-linked immunosorbent assay), and sensitive (LOD<1nM) assay for determination of the DnaK level in cell culture lysates. The operational stability of the method was high compared to that of other SPR assays, the sensitivity decreased at only 2.7%/h. This allowed measurement of more than 220 samples per sensor surface. Storage stability was determined at 25°C (100% after 17h) and 10°C (101% after 1month). The method was validated by standard additions of DnaK (30, 60, and 120nM) with recovery indices in the range 95.7-103.7%. © 2003 Elsevier Inc. All rights reserved.

  • 106.
    Vostiar, Igor
    et al.
    Slovak university of Technology.
    Tkac, Jan
    Slovak Academy of Sciences.
    Mandenius, Carl-Fredrik
    Linköpings universitet, Tekniska högskolan. Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi.
    Off-line monitoring of bacterial stress response during recombinant protein production using an optical biosensor2004Inngår i: Journal of Biotechnology, ISSN 0168-1656, E-ISSN 1873-4863, Vol. 111, nr 2, s. 191-201Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    A surface plasmon resonance (SPR) biosensor was used to monitor the profiles of the heat-shock protein (DnaK) and the expression of a heterologous protein to map the dynamics of the cellular stress response in Escherichia coli. As expression system was used an E. coli strain overproducing human recombinant superoxide dismutase (rhSOD). Expression of DnaK showed complex patterns differing with strength of induction. The strong up-regulation of DnaK expression was observed in all cultivations which over-produced of rhSOD. Similar patterns were not observed in non-induced reference cultures. Differences in DnaK concentration profiles were correlated with induction strength. Presented data, carried out in shake flask and glucose limited fed-batch cultivation, show a good consistency with previously published transcriptional profiling results and provide complementary information to understand stress response related to overproduction of recombinant protein. The study also demonstrates the feasibility of using the SPR as a two channel protein array for monitoring of intracellular components. © 2004 Elsevier B.V. All rights reserved.

  • 107.
    Warth, Benedikt
    et al.
    Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi. Linköpings universitet, Tekniska högskolan.
    Rajkai, Gyorgy
    Belach Biotekn AB.
    Mandenius, Carl-Fredrik
    Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi. Linköpings universitet, Tekniska högskolan.
    Evaluation of software sensors for on-line estimation of culture conditions in an Escherichia coli cultivation expressing a recombinant protein2010Inngår i: JOURNAL OF BIOTECHNOLOGY, ISSN 0168-1656, Vol. 147, nr 1, s. 37-45Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Software sensors for monitoring and on-line estimation of critical bioprocess variables have mainly been used with standard bioreactor sensors, such as electrodes and gas analyzers, where algorithms in the software model have generated the desired state variables. In this article we propose that other on-line instruments, such as NIR probes and on-line HPLC, should be used to make more reliable and flexible software sensors Five software sensor architectures were compared and evaluated. (1) biomass concentration from an on-line NIR probe. (2) biomass concentration from titrant addition, (3) specific growth rate from titrant addition. (4) specific growth rate from the NIR probe, and (5) specific substrate uptake rate and by-product rate from on-line HPLC and NIR probe signals. The software sensors were demonstrated on an Escherichia colt cultivation expressing a recombinant protein, green fluorescent protein (GFP), but the results could be extrapolated to other production organisms and product proteins. We conclude that well-maintained on-line instrumentation (hardware sensors) can increase the potential of software sensors This would also strongly support the intentions with process analytical technology and quality-by-design concepts

123 101 - 107 of 107
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