A biosensor-based phage display selection method for automated, high-throughput Nanobody discovery

Area of Science:

• Biotechnology
• Immunology
• Molecular Biology

Background:

• Traditional Nanobody® (Nb) selection relies on labor-intensive, time-consuming biopanning methods.
• Existing methods often require purified antigens and extensive optimization, increasing costs and duration.

Purpose of the Study:

• To validate an improved biopanning strategy utilizing biosensors for efficient Nanobody® selection.
• To demonstrate the advantages of Octet biolayer interferometry for high-throughput Nanobody® enrichment.
• To reduce the time, labor, and material costs associated with Nanobody® discovery.

Main Methods:

• Phage-displayed Nanobody® libraries were panned against target antigens immobilized on Octet biolayer interferometry sensors.
• Optimization of association/dissociation times and buffer conditions was performed to enhance specificity and affinity.
• Downscaling to a 384-well format and using unpurified targets and phage were explored.

Main Results:

• The Octet panning strategy significantly reduced non-specific and low-affinity binders, decreasing the number of panning rounds required.
• High-affinity Nanobodies® were enriched in as few as three rounds within five days.
• Quantification of binder enrichment was achieved directly via interferometry, eliminating the need for phage titration.

Conclusions:

• Octet biosensor-based panning offers a high-throughput, efficient, and cost-effective alternative to traditional biopanning methods.
• This improved strategy enables rapid isolation of target-specific Nanobodies® using unpurified reagents and simplified quantification.
• The validated method streamlines Nanobody® discovery for subsequent screening and characterization.