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Site-specific DNA-antibody conjugates for specific and sensitive immuno-PCR
- Kazane, Stephanie A., Sok, Devin, Cho, Edward H., Uson, Maria Loressa, Kuhn, Peter, Schultz, Peter G., Smider, Vaughn V.
- Proceedings of the National Academy of Sciences of the United States of America 2012 v.109 no.10 pp. 3731-3736
- amino acids, antibodies, diagnostic techniques, image analysis, labeling techniques, oligonucleotides
- Antibody conjugates are widely used as diagnostics and imaging reagents. However, many such conjugates suffer losses in sensitivity and specificity due to nonspecific labeling techniques. We have developed methodology to site-specifically conjugate oligonucleotides to antibodies containing a genetically encoded unnatural amino acid with orthogonal chemical reactivity. These oligobody molecules were used in immuno-PCR assays to detect Her2+ cells with greater sensitivity and specificity than nonspecifically coupled fragments, and can detect extremely rare Her2+ cells in a complex cellular environment. Such designed antibody-oligonucleotide conjugates should provide sensitive and specific reagents for diagnostics, as well as enable other unique applications based on oligobody building blocks.