Biological Applications of Synthetic Binders Isolated from a Conceptually New Adhiron Library

Area of Science:

• Biotechnology
• Molecular Biology

Background:

• Adhirons are small (10 kDa) synthetic ligands.
• They represent a potential alternative to antibody fragments and other scaffolds like DARPins or affibodies.

Purpose of the Study:

• To develop a novel adhiron phage display library.
• To evaluate the efficacy of this library in isolating high-affinity binders against various antigens.

Main Methods:

• Construction of a new adhiron phage display library incorporating cysteines in hypervariable loops.
• Panning the library against diverse antigens, including plant protoplasts, C-Reactive Protein, and SpyCatcher.

Main Results:

• Isolation of adhirons with low nM binding constants against membrane epitopes and soluble proteins.
• Demonstrated ease of bacterial production (15 mg/L), stability, and low aggregation propensity.
• Successful application of adhirons in ELISA, flow cytometry, Western blot, and electrochemical biosensors.

Conclusions:

• The novel adhiron library is effective in generating high-affinity binders superior to previous adhirons and comparable to nanobodies.
• Isolated adhirons exhibit desirable properties for biotechnological applications and biosensing.