Axxam has many years of experience in the development and use of biochemical assays, having partnered with several pharmaceutical companies in discovery projects. The result is a thorough knowledge of the biochemical requirements for the configuration of enzymatic assays for HTS.
Our expert scientists recombinantly express & purify target proteins using the most advanced techniques, and develop functional assays for proteins belonging to all six enzyme classes as listed below:
Please click on the link below to learn more about biochemical assays at Axxam.
Development of biochemical assays follows a step-wise 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 the biochemical target of your choice - from the isoform of the protein, through the measurement technology, to the final readout. Our streamlined optimization process is tailored to the particular requirements of the final HTS configuration at your facility. This approach has been successful for proteins residing in all areas of the cell - nuclear, cytosolic, various organelles, plasma membrane and those destined for secretion.
We are proud of our 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 rely exclusively on the superior sensitivity of fluorescence-based and luminescence-based readouts. The assays are configured to fully comply with the highly demanding requirements of HTS and they are ready to be run on the screening unit.
We provide enough of the purified enzyme for the HTS screen and 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.
With Axxam’s biochemical assays you get a complete system that will give you the best chance of finding useful starting points for subsequent hit to lead efforts and the support and capabilities you need to move on without delay.
