Figure 1.
a) Nitrocefin assay principle. A change in absorbance at λ=495 nm is measured after the cephalosporin core of nitrocefin is hydrolyzed by β-lactamase b) CCF2 (FRET) assay principle. In the intact CCF2 substrate, coumarin moiety's donor FRET resulting from λ= 409 nm excitation is efficiently quenched by the acceptor fluorescein moiety; hydrolysis of the β-lactam leads to the increase of a donor fluorescence measured at 447 nm and simultaneous decrease of an acceptor fluorescence at measured at 520 nm.