Hybridization assay

Antisense therapy, siRNA, and other oligonucleotide and nucleic acid based biotherapeutics can be quantified with hybridization assays.

Signalling of hybridization methods can be performed using oligonucleotide probes modified in-synthesis with haptens and small molecule ligands which act homologous to the capture and detection antibodies.

As with traditional ELISA, conjugates to horse radish peroxidase (HRP) or alkaline phosphatase (AP) can be used as secondary antibodies.

Thus, optimal salt concentration in hybridization assays varies dependent upon the length and base composition of the analyte, capture and detection probes.

The template probe is fully complementary to the oligonucleotide analyte and is intended to serve as a substrate for T4 DNA ligase-mediated ligation.

Further, more exotic nucleic acid chemistries with oligonucleotide drugs may impact upon the activity of the ligase, which needs to be determined on a case-by-case basis.

In the nuclease hybridization assay, the oligonucleotide analyte is captured onto the solid support such as a 96-well plate via a fully complementary cutting probe.