Selection of binders from phage displayed antibody libraries using the BIAcore biosensor

Area of Science:

• Biotechnology
• Immunology
• Molecular Biology

Background:

• Phage display is a powerful technique for antibody discovery.
• Selection based on affinity is crucial for identifying high-quality antibody candidates.
• Dissociation rate constants are key indicators of antibody-antigen binding stability.

Purpose of the Study:

• To establish a method for selecting phage displayed antibodies based on their dissociation rate constants.
• To demonstrate the utility of BIAcore biosensor technology in this selection process.
• To enable rapid enrichment of high-affinity binders from phage libraries.

Main Methods:

• Utilizing a BIAcore biosensor to immobilize antigen (lysozyme).
• Injecting mixed phage stocks displaying antibodies against specific antigens.
• Eluting bound phages and analyzing for specificity and titer to enrich binders.
• Collecting eluted phages at different time points to correlate with dissociation rates.

Main Results:

• Successful enrichment of phages displaying antibodies specific for lysozyme.
• Demonstrated direct proportionality between elution time and antibody affinity.
• Confirmed that selection is driven by decreased dissociation rate constants.
• Quantified kinetic parameters of expressed soluble antibody fragments.

Conclusions:

• Phage displayed antibodies can be effectively selected based on dissociation rate constants.
• BIAcore biosensor technology provides a rapid and simple approach for this selection.
• This method accelerates the identification of high-affinity antibody binders from phage libraries.