Kinetic measurements of oligodeoxynucleotide hybridization with switchSENSE® and comparison with Biacore and solution data I App Note
Standard multi-purpose biochips can be used to investigate hybridization reactions of oligode-oxynucleotides of any sequence. Functionalization (immobilization) and regeneration is possible using a simple workflow by extending the ligand sequence of interest with an ‘anchor’ sequence which is complementary to the DNA immobilized on the multi-purpose biochip.
The kON, kOFF values determined with the switchSENSE sensor agree very well with SPR measurements performed on a Biacore T200 instrument. The on-rates also agree well with kON values determined by stopped-flow measurements in solution, indicating that surface-induced measurement artifacts did not occur.
The repeatability of the binding kinetics were investigated with regard to spot-to-spot and channel-to-channel variation. Consistent kON and kOFF values were obtained with coefficients of variations of less than 10%.