MONITORING THE HYBRIDIZATION OF THE COMPONENTS OF OLIGONUCLEOTIDE MIXTURES TO IMMOBILIZED DNA VIA MATRIX-ASSISTED-LASER-DESORPTION IONIZATION TIME-OF-FLIGHT MASS-SPECTROMETRY/

Reported here is how the hybridization of individual components of oli gonucleotide mixtures to solid-phase bound complementary strands can b e monitored simultaneously by quantitative matrix-assisted laser desor ption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). Three oligonucleotides, a DNA heptamer, a DNA octamer and a DNA octamer wit h a terminal cholic acid appendage were used as the test mixture. Upon cooling in the presence of a complementary undecamer on controlled po re glass, depletion of the components from the solution was observed, The resulting hybridization curves show the same relative affinities a s traditional UV melting curves with single components and their compl ement, Assays of the kind described here may be used to select high af finity binders from combinatorial libraries of modified antisense olig onucleotides, (C) 1998 John Wiley & Sons, Ltd.