Evaluation of different chemical strategies for conjugation of oligonucleotides to peptides

被引:5
|
作者
Kozlov, Igor A. [1 ]
Melnyk, Peter C. [1 ]
Hachmann, John P. [1 ]
Barker, David L. [1 ]
Lebl, Michal [1 ]
Zhao, Chanfeng [1 ]
机构
[1] Illumina Inc, San Diego, CA 92121 USA
来源
NUCLEOSIDES NUCLEOTIDES & NUCLEIC ACIDS | 2007年 / 26卷 / 10-12期
关键词
oligonucleotide-peptide conjugation; protease assay; kinase assay; proteomics;
D O I
10.1080/15257770701533909
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We developed novel assays for high-throughput detection of one or many kinases or proteases. The assays use hundreds of different Peptide substrates, each covalently linked to an oligonucleotide tag. After incubation with sample, the pool of substrates is hybridized to a microarray containing oligonucleotides complementary to the tag sequences. We screened several specific chemistries for the conjugation based on the following criteria: easy derivatization of oligonucleotides and peptides; high efficiency of the conjugation reaction; good stability of the conjugates; and satisfactory conjugate performance in our assays. We have validated selected method during the successful generation of thousands oligonucteolide-peptide conjugates.
引用
收藏
页码:1353 / 1357
页数:5
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