Construction of Rabbit Immune Antibody Libraries

Area of Science:

• Immunology
• Biotechnology

Background:

• Rabbits offer advantages over mice for generating high-affinity, target-specific antibodies due to stronger immune responses.
• Murine monoclonal antibodies dominate the market due to established hybridoma technology, despite rabbit antibody potential.
• Phage display offers an alternative to hybridoma technology for generating monoclonal antibodies from non-murine species.

Purpose of the Study:

• To describe the construction of a rabbit immune Fab library.
• To facilitate the facile isolation of rabbit monoclonal antibodies.
• To enable rapid identification of high-affinity, target-specific rabbit monoclonal antibodies.

Main Methods:

• Construction of a rabbit immune Fab library from immunized rabbit B cells.
• Amplification of antibody variable domain repertoires and assembly into a Fab repertoire via PCR.
• Cloning Fab genes into a phagemid vector, transformation into E. coli, and rescue of a phage-displayed library.
• Biopanning the phage-displayed library against the immunization antigen.

Main Results:

• Successful construction of a functional rabbit immune Fab library.
• Demonstrated feasibility of isolating target-specific rabbit monoclonal antibodies.
• Potential for rapid identification of high-affinity antibodies.

Conclusions:

• Rabbit immune Fab libraries are a valuable tool for generating high-affinity, target-specific rabbit monoclonal antibodies.
• Phage display selection provides an efficient alternative to hybridoma technology for rabbit antibody discovery.
• This method overcomes limitations associated with murine antibody production.