Click-enabled heterotrifunctional template for sequential bioconjugations.

Area of Science:

• Bioconjugation Chemistry
• Chemical Biology
• Drug Discovery

Background:

• Sequential conjugation of diverse biomolecules is crucial for developing advanced biotherapeutics.
• Existing methods often require multiple steps and purification, limiting efficiency.
• Developing a one-pot system for heterogeneous biomolecule assembly remains a challenge.

Purpose of the Study:

• To develop a heterotrifunctional template for one-pot sequential biomolecule conjugation.
• To demonstrate the versatility of the template with a wide range of molecules.
• To enable the creation of novel biotherapeutics and study complex biological systems.

Main Methods:

• Utilized thiol-maleimide chemistry for initial conjugation.
• Employed both copper-free and copper-mediated click chemistries for subsequent conjugations.
• Performed sequential conjugations in a single reaction vessel (one-pot).

Main Results:

• Successfully conjugated a diverse array of molecules including proteins, peptides, sugars, lipids, fluoroalkanes, biotin, and fluorophores.
• Achieved highly efficient conjugations across all tested substrate types.
• Demonstrated the feasibility of a one-pot sequential conjugation process.

Conclusions:

• The developed heterotrifunctional template offers a versatile and efficient platform for complex biomolecule assembly.
• This method facilitates the creation of chemically-enhanced biotherapeutics, particularly those involving discontinuous epitopes.
• The one-pot approach simplifies conjugation workflows, accelerating research in bioconjugation and drug discovery.