Evaluation of Different Chemical Strategies for Conjugation of Oligonucleotides to Peptides

• Published in: Nucleosides, nucleotides & nucleic acids Vol. 26; no. 10-12; pp. 1353 - 1357
• Main Authors: Kozlov, Igor A., Melnyk, Peter C., Hachmann, John P., Barker, David L., Lebl, Michal, Zhao, Chanfeng
• Format: Journal Article
• Language: English
• Published: United States Taylor & Francis Group 01-01-2007
• 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
• ISSN: 1525-7770; 1532-2335
• DOI: 10.1080/15257770701533909

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.