Raddatz Stefan, Mueller-Ibeler Jochen, Kluge Joachim, Wäss Ludger, Burdinski Gerhard, Havens John R, Onofrey Tom J, Wang Daguang, Schweitzer Markus
Nanogen Recognomics GmbH, Industriepark Höchst G830, 65926 Frankfurt am Main, Germany and. Nanogen Inc., 10398 Pacific Center Court, San Diego, CA 92121, USA.
Nucleic Acids Res. 2002 Nov 1;30(21):4793-802. doi: 10.1093/nar/gkf594.
We report the synthesis of new phosphoramidite building blocks and their use for the modification of oligonucleotides with hydrazides. The reaction of these hydrazide oligonucleotides with active esters and aldehydes is demonstrated for solution conjugation and immobilization. Compared with the established amino modified oligonucleotides, hydrazides show enhanced reactivity at neutral and acidic buffer conditions. One method to introduce hydrazides is using amidites with preformed, protected hydrazides. A completely novel approach is the generation of the hydrazide functionality during the oligonucleotide cleavage and deprotection with hydrazine. Therefore, building blocks for the introduction of esters as hydrazide precursors are described. For the enhanced attachment on surfaces branched modifier amidites, which introduce up to four reactive groups to the oligonucleotide, are applied. The efficiency of branched hydrazide oligonucleotides compared with standard amino modified oligonucleotides for the immobilization of DNA on active electronic Nanogen chips is demonstrated.
我们报道了新型亚磷酰胺砌块的合成及其用于用酰肼修饰寡核苷酸的方法。展示了这些酰肼寡核苷酸与活性酯和醛的反应，用于溶液共轭和固定化。与已有的氨基修饰寡核苷酸相比，酰肼在中性和酸性缓冲条件下表现出更高的反应活性。引入酰肼的一种方法是使用带有预先形成的、受保护的酰肼的亚磷酰胺。一种全新的方法是在寡核苷酸用肼裂解和脱保护过程中生成酰肼官能团。因此，描述了用于引入作为酰肼前体的酯的砌块。为了增强在表面上的附着，应用了分支修饰亚磷酰胺，其可向寡核苷酸引入多达四个反应基团。证明了与标准氨基修饰寡核苷酸相比，分支酰肼寡核苷酸用于将DNA固定在有源电子纳米基因芯片上的效率。
