On silico peptide microarrays for high-resolution mapping of antibody epitopes and diverse protein-protein interactions

Nat Med. 2012 Sep;18(9):1434-40. doi: 10.1038/nm.2913.

Abstract

We developed a new, silicon-based peptide array for a broad range of biological applications, including potential development as a real-time point-of-care platform. We used photolithography on silicon wafers to synthesize microarrays (Intel arrays) that contained every possible overlapping peptide within a linear protein sequence covering the N-terminal tail of human histone H2B. These arrays also included peptides with acetylated and methylated lysine residues, reflecting post-translational modifications of H2B. We defined minimum binding epitopes for commercial antibodies recognizing the modified and unmodified H2B peptides. We further found that this platform is suitable for the highly sensitive characterization of methyltransferases and kinase substrates. The Intel arrays also revealed specific H2B epitopes that are recognized by autoantibodies in individuals with systemic lupus erythematosus who have elevated disease severity. By combining emerging nonfluorescence-based detection methods with an underlying integrated circuit, we are now poised to create a truly transformative proteomics platform with applications in bioscience, drug development and clinical diagnostics.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Computer Systems
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / genetics*
  • Histones / genetics*
  • Humans
  • Methyltransferases / genetics
  • Methyltransferases / metabolism
  • Microarray Analysis / methods*
  • Molecular Sequence Data
  • Peptides / genetics*
  • Phosphorylation
  • Phosphotransferases / genetics
  • Phosphotransferases / metabolism
  • Point-of-Care Systems
  • Protein Interaction Mapping / methods*
  • Proteomics / instrumentation*
  • Proteomics / methods*

Substances

  • Epitopes
  • Histones
  • Peptides
  • Methyltransferases
  • Phosphotransferases