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An inline QC method for determining serial dilution performance of DMSO-based systems.

Leslie A Walling¹

¹Amgen, Inc., M/S 29-1-A, One Amgen Center Drive, Thousand Oaks, CA 91320, USA. lwalling@amgen.com

Journal of Laboratory Automation

|May 26, 2011

Summary

This study introduces Sulforhodamine 101 dye for inline measurement of serial dilutions, improving accuracy in drug candidate efficacy analysis. The new method enhances precision for dose-response curves in dimethylsulfoxide-based systems.

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Area of Science:

Pharmacology and Drug Discovery
Analytical Chemistry
Biotechnology

Background:

Serial dilution is crucial for drug candidate efficacy analysis but prone to error propagation.
Current quality control assumptions in serial dilutions can lead to inaccurate results.
Improving precision and accuracy in dilution plate preparation is essential for reliable dose-response curves.

Purpose of the Study:

To develop an inline performance measurement for enhancing the precision and accuracy of serial dilution protocols.
To introduce Sulforhodamine 101 (S101) dye as a quantitative fluorometric method for dimethylsulfoxide (DMSO)-based systems.
To validate the S101 dye method for quantifying the performance of 96- and 384-well dilution protocols.

Main Methods:

Serial dilution of compounds in DMSO for dose-response curve generation.
Introduction and calibration of Sulforhodamine 101 (S101) dye for inline quantification.
Direct calibration method accounting for S101's nonlinear behavior in DMSO.
Testing of 96- and 384-well dilution protocols on five identical instruments.

Main Results:

S101 dye was selected and validated as a quantitative fluorometric method for DMSO-based systems.
A direct calibration method was developed to address the nonlinear behavior of S101 in DMSO.
The S101 dye method successfully quantified the performance of 96- and 384-well dilution protocols.
The inline measurement approach demonstrated potential for improved precision and accuracy in dilution processes.

Conclusions:

Sulforhodamine 101 dye offers a robust method for inline quantification of serial dilutions in DMSO.
The developed calibration strategy effectively manages S101's nonlinear characteristics.
This approach enhances the reliability of dose-response curve generation and drug candidate efficacy analysis.
The method provides a valuable tool for quality control in high-throughput screening and drug development.