ArticleNucleic acids research2017
DNA-linked Inhibitor Antibody Assay (DIANA) for sensitive and selective enzyme detection and inhibitor screening.
Article in Nucleic acids research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed, 16 citations in OpenAlex.
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- Efficient Method for High-Throughput Screening of Compound Libraries Targeting Human PD-L1.ACS omega · 2026Article
- Identification of 6‑Aryl-7-Deazapurine Ribonucleoside Phosphonates as Inhibitors of Ecto-5'-Nucleotidase (CD73).ACS pharmacology & translational science · 2025Article
- Building the SynBio community in the Czech Republic from the bottom up: You get what you give.Biotechnology notes (Amsterdam, Netherlands) · 2022Article
- Crystal Structure-Guided Design of Bisubstrate Inhibitors and Photoluminescent Probes for Protein Kinases of the PIM Family.Molecules (Basel, Switzerland) · 2021Article
- Activity-Based Diagnostics: An Emerging Paradigm for Disease Detection and Monitoring.Trends in molecular medicine · 2020Review
- DNA-linked inhibitor antibody assay (DIANA) as a new method for screening influenza neuraminidase inhibitors.The Biochemical journal · 2018Article
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Authors and funding
8 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Human diseases are often diagnosed by determining levels of relevant enzymes and treated by enzyme inhibitors. We describe an assay suitable for both ultrasensitive enzyme quantification and quantitative inhibitor screening with unpurified enzymes. In the DNA-linked Inhibitor ANtibody Assay (DIANA), the target enzyme is captured by an immobilized antibody, probed with a small-molecule inhibitor attached to a reporter DNA and detected by quantitative PCR. We validate the approach using the putative cancer markers prostate-specific membrane antigen and carbonic anhydrase IX. We show that DIANA has a linear range of up to six logs and it selectively detects zeptomoles of targets in complex biological samples. DIANA's wide dynamic range permits determination of target enzyme inhibition constants using a single inhibitor concentration. DIANA also enables quantitative screening of small-molecule enzyme inhibitors using microliters of human blood serum containing picograms of target enzyme. DIANA's performance characteristics make it a superior tool for disease detection and drug discovery.
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Registered trials
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