Reagents for biotinylation that are innately water-soluble and can be tracked by both UV-vis spectroscopy and fluorescence (330nm/420nm) have been developed. These reagents are based on the isatoic anhydride platform, which gives rise to their valuable spectroscopic features. The reagents are designed to react quickly with primary and secondary amines, but only sluggishly with neutral water. The reagents snap into place using the spring loaded carbonyl of the anhydride giving CO2 as the only byproduct. A number of reagents have been prepared and tested using a variety of analytical techniques. Biotinylation strategies and thiol reactive reagents will be discussed along with having their utility demonstrated. Asymmetric reagents were generated to show the ease of incorporation of new functionality. The modifications of the isatoic anhydride platform were performed to increase the utility of the reagents for bioconjugation.
