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Target Engagement-Mediated Amplification for Monitoring Drug-Target Interactions in Situ
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- Al-Amin, Rasel A., 1983- (författare)
- Uppsala universitet,Institutionen för immunologi, genetik och patologi,Science for Life Laboratory, SciLifeLab, Science for Life Laboratory, SciLifeLab,Molekylära verktyg, Molecular tools
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- Johansson, Lars (författare)
- Division of Translational Medicine & Chemical Biology, Department of Medical Biochemistry & Biophysics, Karolinska Institutet
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- Landegren, Nils, 1986- (författare)
- Uppsala universitet,Autoimmunitet,Science for Life Laboratory, SciLifeLab,Department of Medicine (Solna), Karolinska University Hospital, Karolinska Institutet,Autoimmunitet, Autoimmunity
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- (creator_code:org_t)
- Engelska.
- Annan publikation (övrigt vetenskapligt/konstnärligt)
Abstract
Ämnesord
Stäng
- It is important to determine the localization of drugs or drug candidates at cellular and subcellular resolution in relevant clinical specimens. This is necessary to evaluate drug candidates from early stages of drug development to clinical evaluation of mutations potentially causing resistance to targeted therapy. We describe a technology where oligonucleotide-conjugated drug molecules are used to visualize and measure target engagement in situ via rolling-circle amplification (RCA) of circularized oligonucleotide probes (padlock probes). We established this target engagement-mediated amplification (TEMA) technique using kinase inhibitor precursor compounds, and we applied the assay to investigate target interactions by microscopy in pathology tissue sections and using flow cytometry for blood samples from patients, as well as in commercial arrays including almost half of all human proteins. In the variant proxTEMAtechnique, in situ proximity ligation assays were performed by combining drug-DNA conjugates with antibody-DNA conjugates to specifically reveal drug binding to particular on- or off-targets in pathological tissues sections. In conclusion, the TEMA methods successfully visualize drug-target interaction by experimental and clinically approved kinase inhibitors in situ and with kinases among a large collection of arrayed proteins.
Nyckelord
- Molecular Biotechnology
- Molekylär bioteknik
Publikations- och innehållstyp
- vet (ämneskategori)
- ovr (ämneskategori)
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