Biomolecular interaction analysis under electrophoretic flow conditions

摘要

Combining the advantages of electrophoresis with the advantages of biomolecular interaction analysis (BIA) enables the biospecific detection of separated molecules; for example it permits differentiation between a complementary single-stranded DNA and a single nucleotide polymorphism. In order to integrate these two techniques, it is necessary to investigate whether it is possible to detect a biomolecular interaction under electrophoretic flow conditions. To this end a novel detection system was developed for electrophoresis that utilizes a label-free and time-resolved detection technique: reflectometric interference spectroscopy (RIfS). The biological functions of important analytes were investigated using this system. Although RIfS can be used as a postcolumn detector, it is also possible to use it to detect relevant substances under electrophoretic flow conditions. DNA–LNA, biotin–streptavidin and protein–protein interactions were detected using this coupled electrophoresis–RIfS set-up.