کد مقاله کد نشریه سال انتشار مقاله انگلیسی ترجمه فارسی نسخه تمام متن
1369416 1379505 2016 3 صفحه PDF ندارد دانلود رایگان
عنوان انگلیسی مقاله
A fluorescence polarization based assay for the identification and characterization of polymerase inhibitors
ترجمه فارسی عنوان
سنجش بر اساس قطب بندی فلورسانس برای شناسایی و توصیف مهارکننده پلی مراز
کلمات کلیدی
پلیمراز؛ سنجش بیوشیمیایی؛ قطب بندی فلورسانس؛ توسعه روش؛ شناسایی ضربه
Polymerases; Biochemical assays; Fluorescence polarization; Assay development; Hit identification
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آلی
چکیده انگلیسی

A homogenous fluorescence polarization (FP) assay was developed to monitor the enzymatic activity of polymerases. Under the optimized conditions established in this study, the assay provides highly robust and reproducible data. Miniaturization of the assay for high-throughput screening and compound testing was also performed. The sensitivity of the newly developed assay was confirmed using 2′,3′-dideoxyadenosine-5′-triphosphate (ddATP), a chain-elongating inhibitor of the polymerase reaction. Side-by-side comparison of the presented fluorescence polarization assay with already well established PicoGreen® fluorescence intensity assay revealed that the performance of both formats is comparable with good assay sensitivity. However, the direct ratiometric readout of the presented FP assay makes it superior over existing colorimetric and fluorescence intensity based assays in terms of susceptibility to false positives. Moreover, due to its generic nature the presented FP assay can be applied to other polymerases and is compatible with identification of inhibitors and requirements of hit-to-lead programs.

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ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Bioorganic & Medicinal Chemistry Letters - Volume 26, Issue 18, 15 September 2016, Pages 4433–4435
نویسندگان
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