Highly efficient selection of epitope specific antibody through competitive yeast display library sorting

Area of Science:

• Immunology
• Biotechnology
• Virology

Background:

• Combinatory antibody library display technologies are crucial for developing therapeutic antibodies.
• Precise epitope targeting is essential for antibodies to effectively modulate molecular interactions.
• Isolating epitope-specific antibodies is vital for various research applications.

Purpose of the Study:

• To develop an efficient strategy for isolating epitope-specific antibodies from yeast display antibody libraries.
• To use dengue virus envelope protein domain III as a model target for antibody selection.
• To validate the isolation of antibodies against a specific, neutralizing epitope.

Main Methods:

• Designed a domain III mutant protein with a key mutation in a neutralizing epitope.
• Employed competitive panning of a yeast display naïve antibody library against wild-type and mutant proteins.
• Sorted and characterized yeast display antibodies that bound to wild-type but not mutant domain III.

Main Results:

• Successfully isolated two unique antibody clones with cross-reactive binding to different dengue virus serotype envelope protein domain IIIs.
• Epitope mapping confirmed one antibody targets the intended neutralizing epitope.
• The method efficiently selected epitope-specific antibodies from a diverse library.

Conclusions:

• The developed strategy enables efficient isolation of epitope-specific antibodies.
• This approach has broad implications for selecting antibodies against conserved viral epitopes and unique cancer targets.
• This method is valuable for engineering therapeutic antibodies with precise targeting capabilities.