Generation of native bovine mAbs by phage display

Area of Science:

• Comparative immunology
• Molecular immunology
• Veterinary medicine

Background:

• Large animals are crucial models for studying diseases relevant to humans and agriculture.
• Current research often relies on murine monoclonal antibodies (mAbs), which can elicit anti-species immune responses in host animals.
• Developing host-derived mAbs is essential for accurate in vivo studies and understanding authentic immune responses in target species.

Purpose of the Study:

• To design and optimize a phage display vector (pComBov) for constructing bovine immunoglobulin (Ig) antigen-binding fragment (Fab) libraries.
• To generate and characterize a panel of bovine mAbs using the novel system.
• To facilitate the use of host-derived mAbs in large animals for comparative immunology and disease research.

Main Methods:

• Design and construction of an optimized phage display vector, pComBov, for bovine Ig Fab libraries.
• Generation of combinatorial libraries of native sequence bovine Ig Fab.
• Production and characterization of bovine mAbs against glutathione S-transferase using ELISA and Western blotting.

Main Results:

• Successfully generated and characterized a panel of bovine mAbs against glutathione S-transferase.
• The isolated bovine mAbs exhibited typical bovine Ig features and high specificity for the target antigen.
• The pComBov system demonstrated effectiveness in producing host-derived monoclonal reagents.

Conclusions:

• The pComBov phage display system is a valuable tool for generating host-derived bovine monoclonal antibodies.
• This technology advances the application of in vivo experiments in large animals by overcoming anti-species responses.
• The system is adaptable for other ruminant species, supporting comparative immunology studies in agriculturally important animals.