Streamlined human antibody generation and optimization by exploiting designed immunoglobulin loci in a B cell line

Area of Science:

• Biotechnology
• Immunology
• Protein Engineering

Background:

• Monoclonal antibodies (mAbs) are crucial therapeutics for cancer, autoimmune, and infectious diseases.
• The avian-derived ADLib system previously enabled rapid generation of antigen-specific mAbs.

Purpose of the Study:

• To develop a human version of the ADLib system for streamlined generation and optimization of functional human mAbs.
• To demonstrate its utility for de novo antibody lead generation and improvement.

Main Methods:

• Construction of tailored human immunoglobulin gene libraries in DT40 cells.
• Isolation of full-length human IgGs against specific antigens.
• Affinity maturation via diversification into secondary libraries and single-cell sorting.

Main Results:

• Successful development of a human ADLib system.
• Demonstrated rapid generation and optimization of functional human mAbs, including antagonistic antibodies.
• Achieved effective affinity maturation, yielding antibodies with improved affinities and functionalities.

Conclusions:

• The human ADLib system is an integrative platform for rapid de novo generation and optimization of therapeutic or diagnostic antibody leads.
• The system's potential extends to directed evolution and optimization of proteins beyond biomedicine.