In the context of biochemistry and drug development, a hybridization assay is a type of Ligand Binding Assay (LBA) used to quantify nucleic acids in biological matrices. Hybridization assays can be in solution or on a solid support such as 96-well plates or labelled beads.
Hybridization assays involve labelled nucleic acid probes to identify related DNA or RNA molecules (i.e. with significantly high degree of sequence similarity) within a complex mixture of unlabelled nucleic acid molecules. 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.