Detergent screening of a G-protein-coupled receptor using serial and array biosensor technologies.

Area of Science:

• Biochemistry
• Biophysics
• Drug Discovery

Background:

• G-protein-coupled receptors (GPCRs) are crucial drug targets.
• Screening solubilization conditions is vital for GPCR research.
• Biosensor technologies offer high-throughput screening solutions.

Purpose of the Study:

• To compare two biosensor platforms for screening GPCR solubilization conditions.
• To evaluate the effectiveness of different detergents on chemokine receptor CCR5 activity.
• To assess the benefits and limitations of serial versus array-based biosensing.

Main Methods:

• Utilized a serial processing instrument (Biacore 2000/3000/T100) and an array platform (Biacore Flexchip).
• Screened 96 detergents for their ability to solubilize CCR5 while maintaining binding activity with a conformationally sensitive Fab (2D7).
• Employed in-line capturing for normalization on the serial platform and off-line capturing for the array platform.

Main Results:

• The serial instrument screened three samples per 30-min cycle, taking ~24h per 96-well plate.
• The array platform screened all 96 detergents simultaneously in under 1h.
• Both platforms identified identical detergents yielding active solubilized CCR5, despite differences in normalization and sensitivity.

Conclusions:

• Serial and array biosensor platforms are complementary for screening GPCR solubilization.
• The array platform offers speed but has limitations in normalization and sensitivity for smaller analytes.
• The serial platform provides robust normalization and higher sensitivity but is slower.