Biomedical Engineering Reference
In-Depth Information
FIGURE 11.9
Benzamidine-based affinity maturation DNA-encoded library. (a) Schematic representation of a library member: a library of 8000
(40
×
200) DNA-tagged benzamidine-based compounds was assembled by sequential stepwise addition of two different sets of building blocks
onto a core structure consisting of (4-carbamimidoylbenzyl)glutamamide; (b) after selection on trypsin-coated resin, high-throughput-sequencing
analysis revealed a strong enrichment of structurally related compounds; (c) table summarizing the inhibition constants of some hits, identified and
resynthesized after trypsin selection. Twenty-one compounds were individually resynthesized as glutamine (Gln) or alanine (Ala) derivatives and
incubated at different concentrations with four different serine proteases (trypsin, thrombin, uPA, and FXa). The reaction was started by addition of the
fluorogenic substrate Z-GGR-AMC and the change of fluorescence signal was recorded. Gln_22_81 exhibited an IC
50
value of 3.0 nM toward trypsin
and excellent specificity toward closely related proteases. (Adapted from [62], with permission; copyright
C
2010 American Chemical Society.)
