EFC Technology - Biochemical format

The biochemical assays are formatted as competitive immunoassays, where analytes are detected in solution, without separation or wash steps, providing improved sensitivity and precision compared to conventional immunoassays. In the biochemical assays, ED is conjugated to ligand of interest (cAMP, cortisol, or cGMP) which competes against free ligand for binding to a binding protein. Depending on the ligand of interest, the binding protein can be an antibody, an enzyme, or a receptor.

In the absence of free ligand, ED-conjugates are captured by the binding protein and are unavailable for complementation, resulting in low signal. In the presence of free ligand, binding protein sites are occupied, leaving ED-conjugate free to complement with EA, forming active EFC β-gal enzyme for substrate hydrolysis to produce a detectable signal. A positive signal is generated in direct proportion to the amount of free ligand bound by the binding protein.

Biochemical assays are referred to as HitHunter™ assays and have been developed for the following target applications:

1.GPCR Second Messenger Signaling
2.Kinase Activity and Binding
3.Nuclear Hormone Receptor Binding
4.Protease Activity