A Guide to Run Affinity Screens Using Differential Scanning Fluorimetry and Surface Plasmon Resonance Assays.

Area of Science:

• Drug discovery and medicinal chemistry
• Biophysics and biochemical assays
• High-throughput screening

Background:

• Drug discovery has shifted from phenotypic to target-based strategies over 30 years.
• High-throughput screening of compound libraries is a primary source for new chemical matter.
• Biophysical methods are crucial for validating hits and ruling out false positives from assay artifacts.

Purpose of the Study:

• To discuss the application of biophysical affinity screening methods in hit identification and validation.
• To highlight advances in throughput, sensitivity, and robustness of these methods.
• To provide practical aspects and caveats for establishing affinity-based screens.

Main Methods:

• Differential scanning fluorimetry (DSF)
• Surface plasmon resonance (SPR)
• Comparison of assay complexity, throughput, and information content.

Main Results:

• Biophysical methods can complement classical functional assays in hit identification.
• Advances allow broader application of affinity screening in drug discovery.
• New target classes, previously inaccessible to functional assays, can now be explored.

Conclusions:

• Differential scanning fluorimetry and surface plasmon resonance are broadly utilized in academia and industry.
• These methods exemplify how diverse assays support hit identification and validation.
• Understanding practical aspects and caveats is key to successful implementation of affinity screens.