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While the miniaturization of the High Throughput Screening (HTS) process has allowed the scaling up of the number of wells/plate from 96 to 384 and up to 1536 and a saving in the amount of compound libraries used for screening, the low number of cells/well means the reporter cells are forced to generate a strong and reproducible signal that can be accurately detected.
Axxam is developing several tools to compensate for the difference between the biopharmaceutical industry's need for low-volume screening and the signal weakness of currently-available tools.
Our researchers focus on the development of new reporter genes and cell lines to overcome these problems, such as:
NORA - An innovative reporter system for detection of sGuanylate Cyclase activity and Nitric Oxide
Axxam scientists have developed an innovative cell-based assay, optimized for use in HTS, for the identification of new modulators of Guanylate Cyclase . This important enzyme catalyzes the biosynthesis of cGMP from GTP.
GC exists in two isoenzyme forms: the soluble and the membrane-bound. The soluble GC (sGC) is a heterodimer composed of an α and a β subunit and it contains heme as a prosthetic group. The most important physiological activator of sGC is nitric oxide (NO), which activates the enzyme upon binding to the heme moiety.
By producing the second messenger cGMP, which regulates various effector systems such as phosphodiesterases, ion channels and protein kinases, sGC plays an important role in different physiological processes thus representing a very attractive pharmacological target.
The innovative luminescent cell-based assay, called NORA (NO Release Assay), is suitable for both 96- and 384-MTP formats and has been developed by stably expressing the α and β subunits of a mutated form of the soluble guanylate cyclase in Chinese Hamster Ovary (CHO) cells. The mutated form synthesizes cAMP, instead of cGMP, allowing the detection of enzymatic activity by a reporter gene approach.
This cell line responds to compounds typically used in the field of sGC research. It represents an innovative and robust assay to screen for sGC modulators with high efficiency and high sensitivity by means of standard luminescence readers.
For NORA licensing opportunities, please contact us.
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