Protein-sensing assay formats and devices

Bilitewski, Ursula

doi:10.1016/j.aca.2005.12.073

Cited by 75 publications

(45 citation statements)

References 76 publications

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“…2) [42,43]. This requires two probes that bind to different regions of the target, yielding enhanced selectivity but increasing development costs and limiting use in research settings.…”

Section: To Label or Not To Label?mentioning

confidence: 99%

Label‐Free Impedance Biosensors: Opportunities and Challenges

2007

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Impedance biosensors are a class of electrical biosensors that show promise for point-of-care and other applications due to low cost, ease of miniaturization, and label-free operation. Unlabeled DNA and protein targets can be detected by monitoring changes in surface impedance when a target molecule binds to an immobilized probe. The affinity capture step leads to challenges shared by all label-free affinity biosensors; these challenges are discussed along with others unique to impedance readout. Various possible mechanisms for impedance change upon target binding are discussed. We critically summarize accomplishments of past label-free impedance biosensors and identify areas for future research.

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“…2) [42,43]. This requires two probes that bind to different regions of the target, yielding enhanced selectivity but increasing development costs and limiting use in research settings.…”

Section: To Label or Not To Label?mentioning

confidence: 99%

Label‐Free Impedance Biosensors: Opportunities and Challenges

2007

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“…Protein adsorption on charged surfaces plays a role in a number of fields such as biomaterials (Wittmer et al, 2007), diagnostics (Bilitewski, 2006), and bioseparations (Khokhlova et al, 2005). Protein adsorption in ion exchange chromatographic systems is of particular importance for the production of biopharmaceuticals.…”

Section: Introductionmentioning

confidence: 99%

Investigation of protein binding affinity and preferred orientations in ion exchange systems using a homologous protein library

Chung¹,

Hou²,

Freed³

et al. 2008

Biotech & Bioengineering

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A library of cold shock protein B (CspB) mutant variants was employed to study protein binding affinity and preferred orientations in cation exchange chromatography. Single site mutations introduced at charged amino acids on the protein surface resulted in a homologous protein set with varying charge density and distribution. The retention times of the mutants varied significantly during linear gradient chromatography. While the expected trends were observed with increasing or decreasing positive charge on the protein surface, the degree of change was a strong function of the location and microenvironment of the mutated amino acid. Quantitative structure-property relationship (QSPR) models were generated using a support vector regression technique that was able to give good predictions of the retention times of the various mutants. Molecular descriptors selected during model generation were used to elucidate the factors affecting protein retention. Electrostatic potential maps were also employed to provide insight into the effects of protein surface topography, charge density and charge distribution on protein binding affinity and possible preferred binding orientations. The use of this protein mutant library in concert with the qualitative and quantitative analyses presented in the article provides an improved understanding of protein behavior in ion exchange systems.

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“…While the binding capacity of protein A is predominantly limited to three human IgG subclasses (IgG 1, 2 and 4), protein G has specificity for subclasses of Abs from many species. In addition, genetic engineering has enabled the production of a fusion protein, protein A/G that combines IgG binding domains of both protein A and protein G. Higher sensing abilities are regularly exhibited for immunoassays employing Ab-binding proteins for Ab immobilisation compared to those using conventional methods such as random covalent immobilisation [29,30]. Although there is the additional step of immobilising the Ab-binding protein on the surface prior to Ab immobilisation, Ab-binding proteins can be genetically engineered and easily prepared in large quantities, providing many options for immobilisation.…”

Section: Antibody-binding Proteinsmentioning

confidence: 99%

Trends and Perspectives in Immunosensors

2012

Antibodies Applications and New Developments

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Immunosensors are devices that comprise both a biomolecular recognition system, such as an antibody and its corresponding antigen, and a transducer to translate the high affinity and specific binding event into a physical signal.Antibodies are produced by an immunological response to the presence of a foreign substance called an antigen. Antibodies may be immobilised onto a variety of platforms including bulk planar surfaces and nanoparticles by either covalent or adsorption strategies. Different interfaces between the bio-components and the detector are available to monitor in 'real-time' the signal generated by biological interactions. The transducers detect, for example, the change in electron transfer, absorbance, fluorescence, refractive index, mass change or heat transfer as the antibody binds to the antigen of interest. Such analytical devices have allowed a wide range of analytes to be identified and quantified such as pathogens, toxins, environmental food contaminants and disease biomarkers.The demand for sensitive, rapid, 'on-site' techniques has taken advantage of the latest advances in microfluidics and nanotechnology. This chapter will highlight current trends in immobilisation, micro/nano-fluidics/ and transducers utilised. A number of examples outlining the exploitation of these elements in immunosensors and their successful applications will be described.

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Protein-sensing assay formats and devices

Cited by 75 publications

References 76 publications

Label‐Free Impedance Biosensors: Opportunities and Challenges

Label‐Free Impedance Biosensors: Opportunities and Challenges

Investigation of protein binding affinity and preferred orientations in ion exchange systems using a homologous protein library

Trends and Perspectives in Immunosensors

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