Multispecific, Multivalent Antibody-Based Molecules Engineered on the DART® and TRIDENT^TM Platforms

Area of Science:

• Biotechnology
• Immunology
• Protein Engineering

Background:

• Conventional monoclonal antibodies have limitations in targeting multiple antigens simultaneously.
• Multispecific antibodies enable advanced therapeutic applications like cell bridging and targeted protein proximity.
• The DART and TRIDENT platforms facilitate the engineering of complex multispecific antibodies.

Purpose of the Study:

• To describe the design and generation of DART and TRIDENT molecules.
• To detail the expression and purification of these engineered antibodies.
• To focus on DART and TRIDENT molecules designed for dual or triple antigen targeting.

Main Methods:

• Engineering of DART (Dual Affinity Re-Targeting) molecules with stabilized diabody-like structures.
• Development of TRIDENT molecules by linking DART units or DART and Fab domains.
• Utilizing Fc domains for half-life modulation and effector function control.
• Expression in CHO cells via transient transfection and subsequent purification.

Main Results:

• Successful design and generation of DART and TRIDENT expression plasmids.
• Expression of engineered multispecific antibodies in CHO cells.
• Purification protocols established for DART and TRIDENT molecules from cell supernatants.
• Demonstrated capability to create molecules with up to tetravalent antigen targeting.

Conclusions:

• DART and TRIDENT platforms provide versatile tools for engineering multispecific antibodies.
• These platforms enable the creation of antibodies with tailored properties for therapeutic applications.
• The described protocols support the development of novel bispecific and trispecific antibodies.