Multiple oligodeoxyribonucleotide syntheses on a reusable solid-phase CPG support via the hydroquinone-O,O '-diacetic acid (Q-Linker) linker arm

A strategy for oligodeoxyribonucleotide synthesis on a reusable CPG solid-p hase support, derivatized with hydroxyl groups instead of amino groups, has been developed. Ester linkages, through a base labile hydroquinone-O,O'-di acetic acid (Q-Linker) linker arm, were used to couple the first nucleoside to the hydroxyl groups on the support. This coupling was rapidly accomplis hed (10 min) using O-benzotriazol-1-yl-N,N,N',N'-tetramethyluronium hexaflu orophosphate (HBTU) and 1-hydroxybenzotriazole as the activating reagents, Oligodeoxyribonucleotide synthesis was performed using existing procedures and reagents, except a more labile capping reagent, such as chloroacetic an hydride, methoxyacetic anhydride or t-butylphenoxyacetic anhydride, was use d instead of acetic anhydride, After each oligodeoxyribonucleotide synthesi s, the product was cleaved from the support with ammonium hydroxide (3 min) and deprotected as usual. Residual linker arms or capping groups were remo ved by treatment with ammonium hydroxide/methylamine reagent and the regene rated support was capable of reuse. Up to six different oligodeoxyribonucle otide syntheses or up to 25 cycles of nucleoside derivatization and cleavag e were consecutively performed on the reusable support. This method may pro vide a significant cost advantage over conventional single-use solid suppor ts currently used for the manufacture of antisense oligodeoxyribonucleotide s.