Protein Function
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Proteomics:  

In the post-genomic era, the study of Protein Function and Structure is paramount to the drug discovery process. About 75% of the all the sequenced human proteins are "orphan" proteins of unknown functions or have poorly defined roles. Many of these orphan proteins are expected to have some catalytic activity, which may or may not have been characterized before.

 

Deciphering of structures and functions of unknown entities (i.e. proteins) is possible by monitoring distinctive thermal signatures of their interaction (binding) with known ligand entities and profiling the quantitative results. Monitoring of such interactions was historically performed manually using individual calorimetric chambers for each of the interactions.

 

Examples of these protein interactions (and functional importance) include:

 

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Protein – protein (pathway analysis);

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Protein – small molecules (drug interaction, structure decoding);

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Protein – peptides, DNA/RNA  (function analysis)
 

The change in Gibb’s Free Energy of the protein-ligand binding interaction can be monitored and expressed by the following; 

DG = DH - T x D S        (H=Enthalpy, S=Entropy and T=Temperature)

 

Applying these principles, TechElan has combined sensitive infrared (IR) imagers with current HTS technologies to present an integrated monitoring system for the high-throughput analysis and profiling of these protein interactions.

 

The system employs sensitive infrared (IR) imagers to directly detect thermal signatures of binding events in a high-throughput fashion using a novel technique of concentrating and registering emitted IR radiation from multiple active and reference binding sites (wells).  The amplified IR signature from each well is analyzed, compared ratiometrically and profiled to a database.

 

Two alternative interaction environments are available for the monitoring of binding events. The IR Plate and the the IR Transmitting Fiber Optics.

 

The IR Plate method is a two-step process, in which entities under investigation are dispensed into individual wells of an IR Plate and then coated with a known ligand by using the Area "Shower" Dispenser. The thermal signatures are monitored by special IR sensors interfaced with the openings in the plate or through IR transmitting fibers.

 

The IR Fiber Optics method is also a two-step process in which tips of an array of individual fibers are first coated by dipping into individual entities under investigation. Then, all fibers are bundled together and dipped into a common and known ligand. Thermal signatures are monitored by an array of individual sensors located at the other terminus of the IR fibers.

 

Both methods of Protein Function and Structure decoding address the sensitive registration of simultaneously triggered binding activities across a large testing array and both methods are reversible in terms of interchanging ligand and unknown entities. Preference of methods is dependent upon assay conditions and automation platforms.

 

General Features of IR Thermography:

 

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A label-free direct detection assay, which minimizes noise and simplifies assay development.  Only a few other techniques (SPR, FFF) can support a label free process.

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Higher resolution of detection, based on the ratiometric (comparative) measurement. 

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Observation of process kinetics (sensogram), which may help in detection of certain non-specific binding activities.

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The ability to "zoom in" or "zoom out" through titration and other methods within the same array.

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Better protein preservation due to less invasive assay environment (as compared with MS, X-ray, and NMR methods).

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The ability to do "coarse" and "fine" investigations moving from protein "clusters/constellations" to purified proteins to protein fractions

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A robotic friendly plate layout suitable for high throughput

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The ability to collect information from a large array (versus sequential testing typical for other protein decoding techniques).

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 Accelerate the drug discovery process by observing direct drug-to-target protein interactions.

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 Broad applications of the technology in Target Screening, Pre-clinical Lead Identification, Clinical - ADME Tox Studies, Pharmaco-genomics and Post-clinical patient monitoring

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High "technological potential" that grows with all thermal, IR Imaging, MEMS and nano-material technologies used in military and astronomy applications.

 

 Please contact us for licensing and other inquiries.