Evaluation of Different Chemical Strategies for Conjugation of Oligonucleotides to Peptides

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 oligon...

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Bibliographic Details
Published in:Nucleosides, nucleotides & nucleic acids nucleotides & nucleic acids, 2007-01, Vol.26 (10-12), p.1353-1357
Main Authors: Kozlov, Igor A., Melnyk, Peter C., Hachmann, John P., Barker, David L., Lebl, Michal, Zhao, Chanfeng
Format: Article
Language:English
Subjects:
DNA - chemistry
kinase assay
Oligonucleotide-peptide conjugation
Oligonucleotides - chemistry
Peptide Hydrolases - analysis
Peptide Hydrolases - chemistry
Peptides - chemistry
Phosphotransferases - analysis
Phosphotransferases - chemistry
protease assay
proteomics
Substrate Specificity
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Summary: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 oligonucleotide-peptide conjugates.
ISSN:1525-7770
1532-2335
DOI:10.1080/15257770701533909