Hydrazino-Pictet-Spengler Ligation as a Biocompatible Method for the Generation of Stable Protein Conjugates

• DOI: 10.1021/bc400042a
• Publication Date: 2013-05-29T00:19:56Z
• Authors: Paresh Agarwal; Romas Kudirka; Aaron E. Albers; Robyn M. Barfield; Gregory W. de Hart; Penelope M. Drake; Lesley C. Jones; David Rabuka
• Keywords: Models, Molecular; 0301 basic medicine; Aldehydes; Molecular Structure; Hydrolysis; Proteins; Biocompatible Materials; Hydrogen-Ion Concentration; Ketones; 03 medical and health sciences; Hydrazines; Oximes; Humans

Aldehyde- and ketone-functionalized biomolecules have found widespread use in biochemical and biotechnological fields. They are typically conjugated with hydrazide or aminooxy nucleophiles under acidic conditions to yield hydrazone or oxime products that are relatively stable, but susceptible to hydrolysis over time. We introduce a new reaction, the hydrazino-Pictet-Spengler (HIPS) ligation, which has two distinct advantages over hydrazone and oxime ligations. First, the HIPS ligation proceeds quickly near neutral pH, allowing for one-step labeling of aldehyde-functionalized proteins under mild conditions. Second, the HIPS ligation product is very stable (>5 days) in human plasma relative to an oxime-linked conjugate (∼1 day), as demonstrated by monitoring protein-fluorophore conjugates by ELISA. Thus, the HIPS ligation exhibits a combination of product stability and speed near neutral pH that is unparalleled by current carbonyl bioconjugation chemistries.