Validated Cell Lines for High-Throughput-Screening
In cooperation with Cisbio Bioassays CCS has developed a set of recombinant cell lines stably expressing G-Protein coupled recptors (GPCRs) of high pharmacological relevance.
The cell lines have been generated by stable transfection of CHO-K1 cells and subsequent clonal selection. They have been specifically validated for a read-out in homogenous time-resolved fluorescence assays (htrf®), measuring second messengers of the different G-Protein couplings.
Cells are supplied either on basis of a licence agreement with the option to propagate the cells in-house or as aliquots of assay-ready Frozen Instant Cells. Each batch of frozen cells has been validated in htrf® assay and has a documented assay window and Z'-factor.
- Validated cell lines for htrf® assays.
- Stability tested for 20 passage doublings.
- Fully automatable in different plate formats.
- Robust assay parameters e.g. S/B, Z' and CV.
- Assay-ready use in suspension as Frozen Instant Cells.
! In some countries there might be restriction for product sale.
! htrf(R) is a registered trade mark of cisbio bioassays.
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CHO-CHRM1
CHO-CHRM1 is a recombinant cell line expressing human muscarinic acetylcholine receptor M1, a target for Alzheimer's disease, schizophrenia and chronic pain. The M1 receptor is signaling via Gq/11 resulting in the activation of the IP3 pathway and a subsequent release of Ca2+ from intracellular stores.
The cell line has been validated for a read-out in htrf® assay measuring IP1, a stabilized degradation product of IP3 (IP-One htrf®) and is a perfect tool for screening agonists, antagonists or modulators of the human M1 receptor. The cells are qualified to be used like a conventional assay reagent and can be added directly in suspension to a prepared compound plate.
Fig.1: Agonist Response
Frozen Instant Cells of CHO-CHRM1 (clone B2) were thawed. 20.000 cells were dispensed into the wells of a white 384-well plate in which agonist was already prepared in serial dilutions. After one hour of incubation the response of the cells was measured in a homogenous time resolved fluorescence assay (IP-ONE htrf® cisbio).
Fig.2: Antagonist Response
Frozen Instant Cells of CHO-CHRM1 (cone B2) were dispensed into an assay plate in which antagonist was already prepared in serial dilutions. After 15' of incubation Oxotremorine M was added to the cells at its EC70. The response of the cells to Oxotremorine M was measured in IP-ONE htrf® assay .
CHO-GPR40 is a recombinant cell line expressing the human free fatty acid receptor 1, a target for diabetes. GPR40/FFAR1 is signaling via Gq/11 resulting in the activation of the IP3 pathway and a subsequent release of Ca2+ from intracellular stores.
The cell line has been validated for a read-out in htrf® assay measuring IP1, a stabilized degradation product of IP3 (IP-One htrf®) and is a perfect tool for screening agonists, antagonists or modulators of the fatty acid receptor 1. The cells are qualified to be used like a conventional assay reagent and can be added directly in suspension to a prepared compound plate.
Fig.1: Agonist Response
Frozen Instant Cells of CHO-GPR40 were thawed. 20.000 cells were dispensed into the wells of a white 384-well plate in which agonist GW 9508 was already prepared in serial dilutions. After one hour of incubation the response of the cells was measured in a homogenous time resolved fluorescence assay (IP-ONE htrf® cisbio).
CHO-GHSR1 is a recombinant cell line expressing human ghrelin receptor, a target for obesity, anorexia and cardiovascular diseases. The ghrelin receptor is signaling via Gq/11 resulting in the activation of the IP3 pathway and a subsequent release of Ca2+ from intracellular stores.
It has been specifically validated for a read-out in a homogenous time-resolved fluorescence assay meassuring IP1, a stabilized degradation product of IP3 (IP-One htrf® - cisbio) and is a perfect tool for screening agonists, antagonists or modulators of the human ghrelin receptor. The cells are qualified to be used like a conventional assay reagent and can be added directly in suspension to a prepared compound plate.
Fig.1: Agonist Response
Different batches of Frozen Instant Cells of CHO-GHSR1 were thawed. 20.000 cells were dispensed into the wells of a white 384-well plate in which agonist was already prepared in serial dilutions. After one hour of incubation the response of the cells was measured in a homogenous time resolved fluorescence assay (IP-ONE htrf® cisbio).
Fig.2: Antagonist Response
Frozen Instant Cells of CHO-GHSR1 were dispensed into 384-well plates in which antagonist was already prepared in serial dilutions. After 15' of incubation Ghrelin was added to the cells at its EC70. The response of the cells to Grehlin was measured in IP-ONE htrf® assay .