Modular generation of multispecific antibodies using protein trans-splicing

Area of Science:

• Biotechnology
• Protein Engineering
• Drug Discovery

Background:

• Multispecific antibodies offer advantages over monoclonal antibodies in drug development.
• The vast design space of multispecific antibodies presents challenges for development.
• Efficient screening platforms are needed to navigate complex antibody designs.

Purpose of the Study:

• To develop a high-throughput platform for producing and screening multispecific antibody libraries.
• To utilize intein-mediated protein splicing for efficient antibody construction.
• To validate the fidelity and applicability of the intein-based splicing approach.

Main Methods:

• Intein-mediated protein splicing for combinatorial antibody library production.
• High-throughput screening of generated multispecific antibody libraries.
• Biochemical and functional characterization of spliced antibodies.
• Integration with the DuetMab platform for complex formats.

Main Results:

• Spliced bispecific antibodies demonstrated comparable biochemical and functional properties to conventionally produced ones.
• Intein splicing successfully generated trispecific antibodies when combined with the DuetMab platform.
• Dual-splicing reactions enabled the creation of bivalent bispecific and trispecific antibody formats.
• The platform proved effective for generating and screening complex multispecific antibody libraries.

Conclusions:

• Intein-based protein splicing is a powerful strategy for efficient multispecific antibody library generation and screening.
• This approach facilitates the rapid identification of optimal antibody configurations and formats.
• The platform accelerates the development of novel antibody-based therapeutics.