From spots to beads-PTM-peptide bead arrays for the characterization of anti-histone antibodies

From spots to beads-PTM-peptide bead arrays for the characterization of anti-histone antibodies
Heubach Y, Planatscher H, Sommersdorf C, Maisch D, Maier J, Joos TO, Templin MF, Poetz O
Proteomics. 2013 Mar;13(6):1010-5. doi: 10.1002/pmic.201200383.

Antibodies that recognize PTMs of histones play a central role in epigenetic proteomic research. Modification-specific antibodies are employed in chromatin immunoprecipitation, for Western blotting and during the immunoprecipitation steps for MS-based global proteomic analyses. Knowledge about the antibodies' off-target binding is essential for the interpretation of experimental data. To address this challenge we developed a fast and cost efficient system for generating peptide bead arrays. We employed this method to establish a bead-based peptide array containing 384 peptides displaying phosphorylated, acetylated, methylated, and citrullinated N-terminal regions of histones H2A, H2B, H3 and H4 and controls. We profiled the binding of 40 PTM-specific antibodies important for epigenetic proteomic research.