Diels-Alder bioconjugation of diene-modified oligonucleotides

被引:74
作者
Hill, KW
Taunton-Rigby, J
Carter, JD
Kropp, E
Vagle, K
Pieken, W
McGee, DPC
Husar, GM
Leuck, M
Anziano, DJ
Sebesta, DP
机构
[1] NeXstar Pharmaceut, Boulder, CO 80301 USA
[2] Proligo LLC, Boulder, CO 80301 USA
关键词
D O I
10.1021/jo0100190
中图分类号
O62 [有机化学];
学科分类号
070303 ; 081704 ;
摘要
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.
引用
收藏
页码:5352 / 5358
页数:7
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