This technology enables extremely fast and stable bioconjugation using a unique boron–nitrogen bond-forming reaction. It works efficiently in water at neutral pH, avoiding harsh conditions required by many existing methods. The approach delivers rapid, clean, and reliable coupling for sensitive biological systems, supporting fast labeling workflows and stable conjugate formation under physiological conditions.
Background: Many bioorthogonal reactions used for biomolecule labeling are slow under physiological conditions or require high reagent concentrations, catalysts, or non-biocompatible environments. These limitations reduce efficiency, increase cost, and restrict applicability in biological systems where mild, rapid, and selective reactions are required. As a result, existing approaches can be unsuitable for sensitive biomolecules or complex biological mixtures. A need exists for a reaction that proceeds quickly at low concentrations, under neutral aqueous conditions, and without catalysts, while maintaining high selectivity and product stability.
Technology Overview: The technology utilizes ortho-formylphenylboronic acid to react with hydrazine-based nucleophiles, forming a boron–nitrogen heterocycle through a two-step process involving rapid hydrazone formation followed by intramolecular cyclization. This reaction proceeds at neutral pH in aqueous solution with very fast kinetics and produces a stable aromatic heterocycle with minimal side products. The resulting conjugates are highly stable and compatible with biological environments, enabling efficient and selective biomolecule coupling. Reaction kinetics measured by absorption spectroscopy show second-order rate constants on the order of 10³ M⁻¹ s⁻¹ with completion within minutes at micromolar concentrations, while product stability has been confirmed with no detectable decomposition over at least 30 days in neutral aqueous buffer.
Advantages: • Achieves reaction rates up to ~1000× faster than traditional hydrazone formation at neutral pH • Forms products with stability exceeding 30 days in physiological buffer without degradation • Operates efficiently at low micromolar reagent concentrations without excess reagents • Completes reactions within minutes under ambient and physiological conditions • Eliminates need for catalysts, acidic pH, or organic solvents • Produces a single dominant product with negligible side reactions • Maintains compatibility with sensitive biomolecules in fully aqueous systems
Applications: • Rapid protein conjugation at low micromolar concentrations in vitro and in cell lysate environments • Biomolecule labeling workflows requiring reaction completion within minutes under physiological pH • Stable drug–biomolecule conjugates with >30 day linker integrity in aqueous storage conditions • Low-dose diagnostic probe labeling where reagent concentrations must remain minimal • Bioconjugation in sensitive biological samples without organic solvents or catalysts • Chemical biology studies requiring fast and selective labeling in complex biological mixtures
Intellectual Property Summary: • United States 10,435,418 Issued 10/8/2019
Stage of Development: Testing done in controlled biochemical systems. Reaction kinetics measured by absorption spectroscopy showing second-order rate constants on the order of 10³ M⁻¹ s⁻¹ and completion within minutes at micromolar concentrations. Product formation and stability confirmed by NMR and spectroscopic analysis demonstrating no detectable decomposition over at least 30 days in neutral aqueous buffer.
Licensing Status: This technology is available for licensing.
Licensing Potential: Strong potential for biotechnology, pharmaceutical, and diagnostics companies seeking rapid, stable, and biocompatible conjugation chemistries for protein labeling, drug conjugates, and chemical biology workflows under physiological conditions.
Additional Information: Additional experimental data, spectroscopic validation results, and application-specific performance details available upon request.
Inventors: Susan Bane, Özlem Dilek, Kamalika Mukherjee