High throughput screening for the design of protein binding polymers

Area of Science:

• Biomaterials Science
• Polymer Chemistry
• Biotechnology

Background:

• Polymer encapsulation enhances protein stability and in vivo half-life.
• Identifying optimal polymer-protein interactions is challenging and resource-intensive.

Purpose of the Study:

• To develop a high-throughput screening method for identifying strong polymer-protein interactions.
• To enable rapid readout of polymer-protein binding using Förster Resonance Energy Transfer (FRET).

Main Methods:

• Iterative screening of 288 polymers with diverse monomer compositions.
• Utilized Förster Resonance Energy Transfer (FRET) for rapid detection of polymer-protein binding.
• Tested against a panel of eight different enzymes to assess binding interactions.

Main Results:

• Optimized assay conditions allowed detection of strong binders at protein concentrations as low as 0.1 μM.
• Identified moderately selective polymer binders and elucidated general polymer design trends for strong binding.
• Observed that binding trends vary across different proteins, highlighting the need for specific screening.

Conclusions:

• The FRET-based screening approach is efficient for identifying polymers for protein encapsulation.
• Successfully applied the method to find lead polymers for TNF-related apoptosis-inducing ligand (TRAIL) encapsulation.
• This technique is valuable for designing polymers for selective protein binding or repulsion, especially with expensive proteins.