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Related Experiment Videos

High throughput screening assay for UDP-glucuronosyltransferase 1A1 glucuronidation profiling.

O V Trubetskoy1, M Finel, M Kurkela

  • 1Quintessence Biosciences, University of Wisconsin, Madison, WI., School of Pharmacy, University of Wisconsin, Madison, WI 53719, USA. vladimir.trubetskoy@mirusbio.com

Assay and Drug Development Technologies
|July 20, 2007
PubMed
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This study introduces a new fluorescence-based high throughput screening (HTS) assay for UDP-glucuronosyltransferase 1A1 (UGT1A1). This assay utilizes polymeric micelles to improve drug-drug interaction prediction and reduce drug development costs.

Area of Science:

  • Biochemistry
  • Pharmacology
  • Drug Development

Background:

  • High throughput screening (HTS) assays are crucial for evaluating drug toxicity and interactions.
  • Existing HTS assays for cytochrome P450s are common, but assays for UDP-glucuronosyltransferases (UGTs) are less developed.
  • UGTs play a significant role in drug metabolism and require robust HTS assay development.

Purpose of the Study:

  • To develop a novel fluorescence-based HTS assay for the UGT1A1 enzyme.
  • To characterize the assay's performance in a 384-well plate format using robotic liquid handling.
  • To identify potential assay modifiers (hits) from a small molecule library.

Main Methods:

  • Development of a fluorescence-based HTS assay for UGT1A1.
  • Utilized recombinant UGT1A1 enzyme and a fluorescent substrate.

Related Experiment Videos

  • Incorporated PreserveX-QML polymeric micelles as a stabilizer and to block nonspecific interactions.
  • Screened a small molecule library using robotic liquid handling in a 384-well plate format.

    • Main Results:

    • Established a functional fluorescence-based HTS assay for UGT1A1.
    • Characterized assay performance metrics suitable for robotic liquid handling.
    • Identified assay modifiers from a small molecule library screen.
    • Evaluated the impact of polymeric micelles on assay performance and compound promiscuity.

    Conclusions:

    • The developed HTS assay is suitable for UGT1A1 characterization and drug-drug interaction studies.
    • Polymeric micelles enhance assay performance and can mitigate compound promiscuity issues.
    • This assay facilitates the development of better drug candidates and reduces drug development costs.