Tag-lite® assay development
Tag-lite® services: building your assays for cell surface receptor study and screening
Based on the Tag-lite technology, Cisbio Bioassays has developed a comprehensive service offer to deliver validated, high quality reagents and turnkey assays for cell surface receptor investigation. All formats and applications may be considered: small molecule screening, biologics screening, lead characterization, etc.
Cisbio offers flexible service packages starting with material design steps such as plasmid construction or stable cell line generation, through to the development of sophisticated biologicals like fluorescent ligands or pre-labeled cell production. Additional services such as full assay development and implementation in your lab are available on request.
- On-demand plasmid generation specific for Tag-lite applications
- Custom ligand labeling available in test or batch sizes
- Stable multimode Tag-lite cell line generation
- Production of pre-labeled Tag-lite cells
- Assay development for ligand binding, receptor dimerization, and internalization
Tag-lite case study: development of an AT2 binding assay
This AT2 binding assay was requested by a customer for a screening campaign. The project was undertaken following a typical 4-phase program: paper study, material and reagent preparation, assay development, delivery.
This phase was carried out free of charge by the Tag-lite technical team at Cisbio Bioassays, in close collaboration with the customer, in order to identify precise assay requirements and acceptance criteria.
After paper study approval, the AT2-SNAP-tag receptor plasmid was generated for optimized expression. Concurrently, the angiotensin peptide was labeled with red acceptor. Ligand synthesis largely depends on the complexity of the chemistry developed for labeling, and therefore drives this step. In this particular case, all basic assay materials could be produced within 5 weeks.
Assay development consisted in generating the HEK293 cell line expressing the AT2 SNAP-tag receptor, followed by optimizing SNAP-Lumi4®-Tb substrate labeling. The binding assay conditions were then optimized and ligand Kd determined. A competition dose response was performed at the Kd with unlabeled AT2 and a reference antagonist (PD123310) as well as a selective antagonist for AT1 receptor (ZD7155).
On the completion of the 11-week project, the AT2 binding assay was delivered to the customer with the following items:
- Tag-lite reagents (pre-labeled cells and labeled AT2 ligand) - 2,500 datapoints (384 low volume format, 20 μL)
- AT2 receptor plasmid - 10 μg
- Comprehensive technical report including assay conditions and protocol, and results obtained during assay development
For the pricing and turn around time estimates for your specific project, please contact your local sales representatives.
|Tag-lite custom plasmid (10 µg)||PLASCUST|
|Tag-lite custom labeled cells||CELLCUST|
|Tag-lite custom plasmid quality control||PLASQC|
|Tag-lite internalization reagent||TAGINT|
|Tag-lite cell line generation||CELLGEN|