Streptavidin Homologues for Applications on Solid Surfaces at High Temperatures

One of the most commonly used bonds between two biomolecules is the bond between biotin and streptavidin (SA) or streptavidin homologs (SAHs). A high dissociation constant and the consequent high-temperature stability even allows for its use in nucleic acid detection under PCR conditions. There are a number of SAHs available and for assay design, it is of great interest which SAH will perform best under assay conditions. Albeit there are numerous single studies on the characterization of SAHs in solution or selected solid-phases, there is no systematic study comparing different SAHs for biomolecule binding, hybridization and PCR assays on solid-phases. We compared streptavidin, core streptavidin, traptavidin, core traptavidin, neutravidin and monomeric streptavidin on the surface of microbeads (10 – 15 µm in diameter). We designed multiplex microbead-based experiments and analyzed simultaneously the binding of biotinylated oligonucleotides, the hybridization of oligonucleotides to complementary capture pr...