A modular approach for assembling aldehyde-tagged proteins on DNA scaffolds

Area of Science:

• Bioconjugation Chemistry
• Molecular Biology
• Immunology

Background:

• Antibody scaffold diversity is crucial for expanding immune system capabilities and developing novel therapeutics.
• Systematic exploration of scaffold diversity requires modular and site-specific protein-nucleic acid conjugation methods.
• Current methods lack the simplicity and modularity needed for efficient protein-DNA assembly.

Purpose of the Study:

• To develop a modular and site-specific conjugation strategy for linking synthetic oligonucleotides to proteins.
• To enable the assembly of protein-DNA conjugates onto DNA-based scaffolds with precise spatial control.
• To facilitate the exploration of expanded scaffold diversity for immunoglobulin-based probes and therapeutics.

Main Methods:

• Utilized aldehyde tags on proteins for conjugation to synthetic oligonucleotides.
• Developed a modular approach for site-specific protein-nucleic acid conjugation.
• Assembled the resulting protein-DNA conjugates onto DNA-based scaffolds with nanometer-scale resolution.

Main Results:

• Successfully conjugated synthetic oligonucleotides to proteins with aldehyde tags.
• Demonstrated assembly of protein-DNA conjugates onto DNA scaffolds with low nanometer spatial resolution.
• Confirmed that the resulting conjugates can bind to live cells.

Conclusions:

• The developed modular and site-specific conjugation strategy is a valuable tool for protein-DNA assembly.
• This method facilitates the exploration of expanded antibody scaffold diversity.
• The approach has potential applications in developing enhanced therapeutics and biological probes.