INFLUENCE OF ELUENT ANIONS IN BORONATE AFFINITY-CHROMATOGRAPHY

The influence of various anions on the retention of nucleosides and nu cleotides to a boronate affinity chromatography column was examined. A t pH 7.0, F- ions were found to produce the strongest affinity and the best separation of nucleosides from their 2/-deoxy analogues. B-11 NM R studies indicated this was due in part to the propensity of F- to fo rm tetrahedral fluoroboronate complexes. Using an eluent of 50 mM 4-(2 -hydroxyethyl)-1-piperazineethanesulfonic acid and 500 mM KF at pH 7.0 , the nucleotides nicotinamide-adenine dinucleotide and flavin-adenine dinucleotide were found to be strongly retained on a boronate column. These nucleotide-saturated columns were examined for their ability to retain the nucleotide-binding enzymes, glucose-6-phosphate dehydroena se, lactate dehydrogenase and galactose oxidase. No retention of the e nzymes was observed.