Academic Journal
DNA-linked inhibitor antibody assay (DIANA) as a new method for screening influenza neuraminidase inhibitors
| العنوان: | DNA-linked inhibitor antibody assay (DIANA) as a new method for screening influenza neuraminidase inhibitors |
|---|---|
| المؤلفون: | Kožíšek, Milan, Navrátil, Václav, Rojíková, Kateřina, Pokorná, Jana, Berenguer Albiñana, Carlos, Pachl, Petr, Zemanová, Jitka, Machara, Aleš, Šácha, Pavel, Hudlický, Jason, Císařová, Ivana, Řezáčová, Pavlína, Konvalinka, Jan |
| المصدر: | Biochemical Journal ; volume 475, issue 23, page 3847-3860 ; ISSN 0264-6021 1470-8728 |
| بيانات النشر: | Portland Press Ltd. |
| سنة النشر: | 2018 |
| الوصف: | Influenza neuraminidase is responsible for the escape of new viral particles from the infected cell surface. Several neuraminidase inhibitors are used clinically to treat patients or stockpiled for emergencies. However, the increasing development of viral resistance against approved inhibitors has underscored the need for the development of new antivirals effective against resistant influenza strains. A facile, sensitive, and inexpensive screening method would help achieve this goal. Recently, we described a multiwell plate-based DNA-linked inhibitor antibody assay (DIANA). This highly sensitive method can quantify femtomolar concentrations of enzymes. DIANA also has been applied to high-throughput enzyme inhibitor screening, allowing the evaluation of inhibition constants from a single inhibitor concentration. Here, we report the design, synthesis, and structural characterization of a tamiphosphor derivative linked to a reporter DNA oligonucleotide for the development of a DIANA-type assay to screen potential influenza neuraminidase inhibitors. The neuraminidase is first captured by an immobilized antibody, and the test compound competes for binding to the enzyme with the oligo-linked detection probe, which is then quantified by qPCR. We validated this novel assay by comparing it with the standard fluorometric assay and demonstrated its usefulness for sensitive neuraminidase detection as well as high-throughput screening of potential new neuraminidase inhibitors. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| DOI: | 10.1042/bcj20180764 |
| الاتاحة: | http://dx.doi.org/10.1042/bcj20180764 https://portlandpress.com/biochemj/article-pdf/475/23/3847/733699/bcj-2018-0764.pdf |
| Rights: | https://creativecommons.org/licenses/by-nc-nd/4.0/ |
| رقم الانضمام: | edsbas.41C33938 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1042/bcj20180764 |
|---|