Figure 2.

Fragment-based screening of CCL28 by 2D NMR. Overlays of CCL28(1-108) (100 μM) HSQC spectra in 3% v/v d₆-DMSO (green contours) and a 12-plex mixture of compounds (0.5 mM each) from a Maybridge fragment library (magenta) for representative (A) non-hit (MB07) and (B) hit (MH06) samples. (C) PCA analysis of all 12-plex HSQC spectra, colored according to the result of k-means clustering. Hits are more likely found in cluster 2 or 3 than in cluster 1, which includes the reference spectrum (black star). (D) HSQC difference plots (reference spectrum is subtracted from the 12-plex spectrum) for non-hit and hit samples shown in panels (A) and (B). (E) DIA plot for all 12-plex samples in the Maybridge library screen, sorted by magnitude within k-means clusters from PCA analysis shown in panel C. (F) Summary of the primary screen of CCL28(1-108) against fragment libraries from Zenobia, Maybridge, Enamine and Targetmol, with a representative set of 12-plex, 3-plex and single compound HSQC difference plots shown for each sample leading to identification of an individual fragment hit (7H04).