Bioconjugation and Protein Labeling with Photo-Initiated, Bench-Stable, Water-Soluble Triazene Reagents

Triazenes and methods of producing diazonium species from said triazenes using ultraviolet (UV) light, which provide a fast, easy, stable, scalable, and selectively-triggerable means of modifying aromatic nucleophiles, including those on protein surfaces. The invention includes triazenes for use as bioconjugates, e.g., for use in protein modification, for use as probes (including but not limited to detectable probes such as fluorescent probes), protein crosslinking, etc. Application of UV light at neutral pH releases diazonium species, which can be used in a wide variety of applications, including protein tagging or modification. This chemistry may be used to create libraries of designed conjugates that may be stored safely for long periods of time.

Background:
Chemical labeling of proteins is important for understanding protein structure and function. Labeling natural amino acids in protein of interest is one approach to reach this goal. Most of current methods involve harsh conditions which causes protein denaturation or have low reaction yield. The technology intends to improve on these shortcomings through high-yield and better stability. The relatively low prevalence and absence of charge allows for a more controlled labeling without significantly affecting protein stability.

Applications:

• Protein labeling
• Antibody drug conjugates
• Labeling materials with electron-rich aromatics
• Fluorescent probes
• Chimeras where two proteins are covalently linked
• Modification of tyrosine, histidine, or tryptophan residues
• Create libraries of designed conjugates for stable storage

Advantages:

• Activated by UV light
• Works at neutral pH
• Fast reactions in seconds to minutes
• More controlled labeling
• Scalable, easy to synthesize
• Stable at room temperature