Antibody dual-functionalisation enabled through a modular divinylpyrimidine disulfide rebridging strategy

Area of Science:

• Bioconjugation Chemistry
• Antibody Engineering
• Chemical Biology

Background:

• Antibody functionalization is crucial for developing targeted therapies and diagnostics.
• Existing methods often require metal catalysts or multiple purification steps, increasing complexity and cost.

Purpose of the Study:

• To develop a streamlined, metal-free methodology for the dual-functionalization of IgG antibodies.
• To enable sequential bioorthogonal reactions for precise antibody modification.

Main Methods:

• Utilized disulfide rebridging with divinylpyrimidine technology.
• Incorporated bicyclononyne and methylcyclopropene handles for sequential strain-promoted azide-alkyne cycloaddition (SPAAC) and ইনস্ট্যান্ট ইলেক্ট্রন ট্রান্সফার ডাইঅ্যাজোমেথেন অ্যাডিকেশন (IEDDA) reactions.
• Avoided metal catalysis and intermediate purification steps.

Main Results:

• Successfully achieved dual-functionalization of IgG antibodies in a single workflow.
• Demonstrated the compatibility of the method with sequential SPAAC and IEDDA reactions.
• Validated a metal-free and purification-free approach for antibody modification.

Conclusions:

• The developed methodology offers an efficient and versatile platform for IgG antibody dual-functionalization.
• This metal-free, purification-free strategy simplifies antibody engineering for various applications.
• Paves the way for more accessible and scalable antibody-based therapeutic and diagnostic development.