Team:Bielefeld-Germany/Future

From 2011.igem.org

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The reviewed molecular beacon based NAD<sup>+</sup> bioassay can be applied to biochemical and biomedical studies. Accordingly, it can be utilized to detect NAD<sup>+</sup>/NADH-dependent enzymatic processes. The low limit of detection, its reliability and manageability provides a practical alternative to present colorimetric, fluorometric, chemiluminescent, electrochemical or mass spectrometric methods detecting NAD<sup>+</sup> or NADH. In context of clinical applications and therapeutics the NAD<sup>+</sup> bioassay can be useful to monitor cellular NAD<sup>+</sup> levels during treatments of tissues or cell cultures for identification of drug targets, for instance. It may also be applied in the field of diagnostics. Finally, the principle of the proposed NAD<sup>+</sup> bioassay can be used for cell-free optical biosensors by taking molecular beacons either immobilized on the sensor surface or being present in the reaction medium.
The reviewed molecular beacon based NAD<sup>+</sup> bioassay can be applied to biochemical and biomedical studies. Accordingly, it can be utilized to detect NAD<sup>+</sup>/NADH-dependent enzymatic processes. The low limit of detection, its reliability and manageability provides a practical alternative to present colorimetric, fluorometric, chemiluminescent, electrochemical or mass spectrometric methods detecting NAD<sup>+</sup> or NADH. In context of clinical applications and therapeutics the NAD<sup>+</sup> bioassay can be useful to monitor cellular NAD<sup>+</sup> levels during treatments of tissues or cell cultures for identification of drug targets, for instance. It may also be applied in the field of diagnostics. Finally, the principle of the proposed NAD<sup>+</sup> bioassay can be used for cell-free optical biosensors by taking molecular beacons either immobilized on the sensor surface or being present in the reaction medium.
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[https://2011.igem.org/Team:Bielefeld-Germany/Project#NAD.2B_detection Read a more detailed NAD<sup>+</sup> detection description]
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[https://2011.igem.org/Team:Bielefeld-Germany/Project/Description#NAD.2B_detection Read a more detailed NAD<sup>+</sup> detection description]
===Impact on the Parts Registry===
===Impact on the Parts Registry===

Revision as of 22:45, 28 October 2011

Contents

S-layer

Applications

The ability of self-assembly and forming defined nanostructures enables the use of S-layer proteins as building blocks in a biomolecular construction kit for the generation of a universal nanobiotechnological matrix. By using the matrix as a nano pinboard for further components or just the nanostructure itself, a great variety of applications in modular nanobiotechnology, biomimetics, bioanalytics and medical diagnostic are imaginable.

Concentrating on the repetitive features of S-layer proteins to form a well defined nano lattice, the use as filtration units become reasonable. So-called isoporous S-layer ultrafiltration membrane (SUM) combine the advantages of a constant lattice with a well-defined pore size and the presence of functional groups in defined positions and orientations which are a common problem of current ultrafiltration membranes.

A current topic in S-layer research is the design of more sensitive and improved optical and electrochemical sensors. One example is the luminescence lifetime based oxygen sensor. The key benefits of using S-layer proteins for this type of sensor are the imparted anti-fouling properties and the good biocompatibility. This is especially valuable for sensing applications in complex biological fluids or implantable sensors.

The self-assembly and immobilization properties of S-layer fusion proteins also open new innovative perspectives in clinical applications like immune therapy, blood purification and drug targeting. The genetic fusion of an allergen to SbsC S-layer proteins from Geobacillus stearothermophilus combine reduced allergenicity with immunomodulatory capacity. These qualities make them to an ideal allergen carrier/adjuvants in specific immunotherapy, which is the only causative treatment for type I allergy. In conclusion, S-layer proteins are one of the most innovative and promising discoveries for combining biology with engineering in the new field of nanobiotechnology.

Read a more detailed S-layer description

Impact on the Parts Registry

With our approach in the field of functional S-layer proteins we made it easy for future iGEM teams to create compact, well defined nanobiotechnological surfaces with chosen characteristics and to realize their own projects in a cell-free and biosafe way.

We provided two different fully BioBrick-compatible S-layer genes to the partsregistry (<partinfo>K525301</partinfo> and <partinfo>K525401</partinfo>) which cover the lattice geometries oblique and square, so future iGEM-Teams are able to choose the one they like to use for their application. Because of the [http://partsregistry.org/Assembly_standard_25 Freiburg assembly standard] it is easy to create fusion proteins with the S-layer self-assembly domain on the one side and a functional domain on the other side.

Take a look at our practical guide to do it yourself nanobiotechnology to plan and realize your project with our S-layer proteins.

NAD+ detection

Applications

The reviewed molecular beacon based NAD+ bioassay can be applied to biochemical and biomedical studies. Accordingly, it can be utilized to detect NAD+/NADH-dependent enzymatic processes. The low limit of detection, its reliability and manageability provides a practical alternative to present colorimetric, fluorometric, chemiluminescent, electrochemical or mass spectrometric methods detecting NAD+ or NADH. In context of clinical applications and therapeutics the NAD+ bioassay can be useful to monitor cellular NAD+ levels during treatments of tissues or cell cultures for identification of drug targets, for instance. It may also be applied in the field of diagnostics. Finally, the principle of the proposed NAD+ bioassay can be used for cell-free optical biosensors by taking molecular beacons either immobilized on the sensor surface or being present in the reaction medium.

Read a more detailed NAD+ detection description

Impact on the Parts Registry

With our provided NAD+ dependent Ligase (<partinfo>K525710</partinfo>) in the combination with a molecular beacon, we made it possible for future iGEM teams to detect and meassure any NAD+ producing reaction. As shown in the results the detection is highly sensitive and selective.

By fusing your NAD+ producing enzymes to our provided S-layer proteins (<partinfo>K525301</partinfo> and <partinfo>K525401</partinfo>), it is possible to combine these fusion protein with our provided S-layer fused Ligase (<partinfo>K525005</partinfo> and <partinfo>K525006</partinfo>) to create a nanobiotechnological surface and to establish a biosensor with close proximity of the detecting and the indicating part.