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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:
New Ca2+ activated photoproteins with higher light generation
The bivalent ion Ca2+ is a critical second messenger involved in many physiological and signal transduction processes within a cell. Transient changes in intracellular Ca2+ concentrations are mainly triggered either by ligand-mediated activation of GPCRs or by opening of Ca2+ permeable ion channels. Both are transmembrane proteins and belong to the most preferred target classes within the pharmaceutical industry.
The development of precise Ca2+ mobilisation assays for HTS applications is challenged by the nature of the Ca2+ signal which is rapid, short, transient and sometimes weak. To face these challenges, Axxam scientists developed and characterized a new Ca2+ activated photoprotein called Photina® and validated its use in mammalian cell lines for setting up flash luminescent cell-based assays for HTS applications.
The use of a Ca2+ activated photoprotein with its inherent property of flash-type
luminescence represents a valid tool for analysing all aspects of Ca2+ mediated signal transduction processes in mammalian cells. The advantages of using photoproteins as Ca2+ indicators in HTS include:
- low background levels result in a large signal to noise ratio
- Ca2+ concentrations can be measured at specific cellular sites
- tested compounds only require short incubation periods
- reaction kinetics can be followed. .
Photina® - a new and improved Ca2+ activated photoprotein - was developed by the Axxam Enabling Technologies group. Learn more about Photina® in our Discovery Services and Products section.
In addition to Photina®, Axxam researchers are engaged in finding and developing new photoproteins with novel or improved properties, such as wavelength, Ca2+ affinity etc.
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