Sheep monoclonal antibody fragments generated using a phage display system

Area of Science:

• Immunology
• Biotechnology
• Molecular Biology

Background:

• Monoclonal sheep antibodies hold significant promise for biomedical, veterinary, and agricultural applications.
• Conventional methods for generating sheep monoclonal antibodies, such as hybridoma technology, are not routine and can be complex.

Purpose of the Study:

• To establish an efficient method for generating recombinant sheep antibody fragments from immunized animals.
• To develop a valuable tool for studying sheep immunology and for general biorecognition.

Main Methods:

• Utilized a modified phage display system to generate recombinant antibody fragments.
• Amplified immunoglobulin V gene repertoires from the spleens of sheep immunized with human serum albumin and conalbumin.
• Constructed a phage display single-chain variable fragment (scFv) library using an efficient two-step cloning method.

Main Results:

• Successfully isolated and characterized 14 different single-chain Fv (scFv) fragments.
• Sequence analysis confirmed typical ovine immunoglobulin characteristics, identifying thirteen Vlambda and 11 VH genes.
• Produced soluble monomeric scFvs in Escherichia coli and measured affinities using surface plasmon resonance, observing affinities typical of a secondary immune response.

Conclusions:

• The described method provides a robust approach for generating recombinant sheep antibody fragments.
• This technique is valuable for advancing sheep immunology research and facilitating biorecognition studies.
• The generated scFvs exhibit characteristics suitable for various biotechnological applications.