title: Diels--Alder bioconjugation of diene-modified oligonucleotides.
abstract: In an effort to offer complementary technology for covalent biomolecule modification (bioconjugation), we have developed a method that exploits the aqueous acceleration of Diels--Alder reactions for this purpose. Three different diene phosphoramidite reagents have been synthesized that enable diene modification of synthetic oligonucleotides prepared by the phosphoramidite method. Clean and efficient Diels--Alder cycloaddition of these diene oligonucleotides with maleimide dieneophiles was carried out, and the labeled oligonucleotide bioconjugates were characterized by HPLC and electrospray mass spectrometry. Dieneophile stoichiometry, temperature, and pH are all parameters that were shown to influence the efficiency of the process.
authors.affiliation: NeXstar Pharmaceuticals, 2860 Wilderness Place, Boulder, Colorado 80301, USA. dsebesta@proligo.com
authors.forename: K W
authors.initials: KW
authors.lastname: Hill
authors.forename: J
authors.initials: J
authors.lastname: Taunton-Rigby
authors.forename: J D
authors.initials: JD
authors.lastname: Carter
authors.forename: E
authors.initials: E
authors.lastname: Kropp
authors.forename: K
authors.initials: K
authors.lastname: Vagle
authors.forename: W
authors.initials: W
authors.lastname: Pieken
authors.forename: D P
authors.initials: DP
authors.lastname: McGee
authors.forename: G M
authors.initials: GM
authors.lastname: Husar
authors.forename: M
authors.initials: M
authors.lastname: Leuck
authors.forename: D J
authors.initials: DJ
authors.lastname: Anziano
authors.forename: D P
authors.initials: DP
authors.lastname: Sebesta
mesh_terms: D019921:Biotinylation; D003374:Coumarins / Q000737:chemistry; D019793:Fluorescein / Q000737:chemistry; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D013696:Temperature
chemical_list: D003374:Coumarins; D009841:Oligonucleotides; C030123:coumarin; D019793:Fluorescein