Tag-lite® full assay development

From plasmid design and receptor expression optimization to ligand generation, Cisbio will develop a ready-to-use binding kit for you.

Humans count over 300 different GPCRs. Binding assays for many of these GPCRs have been developed and are readily available from Cisbio's current catalog. GPCRs for which a Tag-Lite binding assay is not yet available may be addressed through Cisbio services

Project definition and feasibility

This section identifies your precise assay requirements and acceptance criteria. Feasibility is evaluated at no cost.

Development

Deliverables

At project completion, Cisbio delivers a fully optimized, ready-to-use binding assay.

Case study : Development of an Angiotensin II receptor ligand 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: This phase was carried out free of charge, in close collaboration with the customer, in order to identify precise assay requirements and acceptance criteria.

Material preparation: After paper study approval:

  • The AT2-SNAP-tag receptor plasmid was generated
  • The angiotensin peptide* was labeled with red acceptor.
  • *Ligand synthesis depends on the complexity of the chemistry developed for labeling

Assay development consists in:

  • Generating the HEK293 cell line expressing the AT2 SNAP-tag receptor
  • Optimizing SNAP-Lumi4®-Tb substrate labeling
  • Optimizing the binding assay conditions
  • Determining ligand Kd determined
  • Performing Ki determination on unlabeled AT2, reference antagonist (PD123310), selective antagonist for AT1 receptor (ZD7155).

Assay delivery, the AT2 binding assay was delivered to the customer with Tag-lite reagents - cells, ligand and plasmid – and a comprehensive technical report including assay conditions and protocol, and results obtained during assay development.