Biomolecular interaction analysis under electrophoretic flow conditions.
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.
Kumpf
Michael
M
Institute of Physical and Theoretical Chemistrym, IPTC, Eberhard-Karls-University of Tuebingen, Auf der Morgenstelle 8, 72076 Tuebingen, Germany. Michael.kumpf@ipc.uni-tuebingen.de
Gauglitz
Günter
G
D000906:Antibodies / Q000032:analysis*; D001710:Biotin / Q000032:analysis*; D004247:DNA / Q000032:analysis*; D004586:Electrophoresis / Q000379:methods; D008857:Microscopy, Interference / Q000379:methods*; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense / Q000032:analysis*; D020641:Polymorphism, Single Nucleotide; D012680:Sensitivity and Specificity; D019809:Streptavidin / Q000032:analysis*; D013997:Time Factors
D000906:Antibodies; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; C477371:locked nucleic acid; D001710:Biotin; D004247:DNA; D019809:Streptavidin