title: Release of DNA oligonucleotides and their conjugates from controlled-pore glass under thermolytic conditions.
abstract: The sequential functionalization of long-chain alkylamine controlled-pore glass (CPG) with a 3-hydroxypropyl-(2-cyanoethyl)thiophosphoryl linker and a dinucleoside phosphorotetrazolide leads to a uniquely engineered support for solid-phase synthesis. Unlike conventional succinylated-CPG supports, this support is designed to allow oligonucleotide deprotection and elimination of deprotection side-products to proceed without release of the oligonucleotide. When needed, the DNA oligonucleotide can be thermolytically released in 2 hr under essentially neutral conditions. The modified CPG support has been successfully employed in the synthesis of both native and fully phosphorothioated DNA 20-mers. On the basis of reversed-phase HPLC and electrophoretic analyses, the purity of the released oligonucleotides is comparable to that of identical oligonucleotides synthesized from succinylated-CPG supports, in terms of both shorter-than-full-length oligonucleotide contaminants and overall yields. The detailed preparation of DNA oligonucleotides conjugated with exemplary reporter or functional groups, either at the 3'-terminus or at both 3'- and 5'-termini, is also described.
authors.affiliation: Division of Therapeutic Proteins, Center for Drug Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, USA.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Grajkowski
authors.forename: Jacek
authors.initials: J
authors.lastname: Cieślak
authors.forename: Scott
authors.initials: S
authors.lastname: Norris
authors.forename: Darón I
authors.initials: DI
authors.lastname: Freedberg
authors.forename: Jon S
authors.initials: JS
authors.lastname: Kauffman
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Duff
authors.forename: Serge L
authors.initials: SL
authors.lastname: Beaucage
mesh_terms: D003432:Cross-Linking Reagents; D005898:Glass; D006358:Hot Temperature; D008024:Ligands; D008722:Methods; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D013499:Surface Properties
chemical_list: D003432:Cross-Linking Reagents; D008024:Ligands; D009838:Oligodeoxyribonucleotides