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The Axxam Biochemistry Group has many years of experience partnering with pharmaceutical companies in drug discovery projects. The result is a thorough knowledge of the biochemical requirements for the configuration of enzymatic assays for high-throughput screening.
Our scientists recombinantly express, purify and develop functional assays for proteins belonging to all enzyme
classes:
- Oxidoreductases
- Isomerases
- Ligases
- Transferases, with a focus on Protein Kinases
- Hydrolases, with a strong competence for Lipases and Proteases
Development of biochemical assays follows a sequential process resulting in the maximal optimization of enzyme activity. Several conditions are screened in parallel along the entire process – from protein expression to assay configuration.
Our scientists design a specific strategy for each biochemical target - from the expression version to the final readout. Our streamlined optimization process is tailored to the particular requirements of the final HTS configuration. This approach has been successful for proteins residing in all areas of the cell - nuclear and cytosolic enzymes and proteins with targeted secretion to various organelles, to the plasma membrane and to the secretory pathway.
We are proud of our past successes in assay configuration for some challenging proteins such as integral transmembrane enzymes and heteromultimeric complexes with up to four essential subunits.
We know that the results of an HTS campaign are inherently dependent on the quality of the assay set up. Therefore, our efforts are directed towards the configuration of functional assays based on the catalytic activity of the target, where the reaction conditions are optimized to mimic the physiological requirements of the enzyme in terms of substrates, regulators and cofactors.
The assays must satisfy the highest standards for HTS, with an emphasis on:
- Signal-to-background ratio
- Standard deviation
- Z’ factor
- Stability of the reaction over the screening time
- Consistent automation
- Miniaturization up to 1536-MTP format
All assays are based on homogeneous addition-only steps and they exclusively rely on the superior sensitivity of fluorescence-based and luminescence-based readouts. This means the assays are configured to fully comply with the highly demanding requirements of HTS and they are ready to be adapted and run on the screening unit. We also provide the entire batch of the purified enzyme to support the follow-on screening and hit confirmation. Depending on the specific activity of the enzyme, the total quantity supplied may correspond to several milligrams of purified enzyme. |
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