Antigen clasping by two antigen-binding sites of an exceptionally specific antibody for histone methylation

Proc Natl Acad Sci U S A. 2016 Feb 23;113(8):2092-7. doi: 10.1073/pnas.1522691113. Epub 2016 Feb 9.


Antibodies have a well-established modular architecture wherein the antigen-binding site residing in the antigen-binding fragment (Fab or Fv) is an autonomous and complete unit for antigen recognition. Here, we describe antibodies departing from this paradigm. We developed recombinant antibodies to trimethylated lysine residues on histone H3, important epigenetic marks and challenging targets for molecular recognition. Quantitative characterization demonstrated their exquisite specificity and high affinity, and they performed well in common epigenetics applications. Surprisingly, crystal structures and biophysical analyses revealed that two antigen-binding sites of these antibodies form a head-to-head dimer and cooperatively recognize the antigen in the dimer interface. This "antigen clasping" produced an expansive interface where trimethylated Lys bound to an unusually extensive aromatic cage in one Fab and the histone N terminus to a pocket in the other, thereby rationalizing the high specificity. A long-neck antibody format with a long linker between the antigen-binding module and the Fc region facilitated antigen clasping and achieved both high specificity and high potency. Antigen clasping substantially expands the paradigm of antibody-antigen recognition and suggests a strategy for developing extremely specific antibodies.

Keywords: antibody engineering; antibody validation; data reproducibility; epigenetics; protein–protein interaction.

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

  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / genetics
  • Antigens / chemistry*
  • Antigens / genetics
  • Binding Sites, Antibody*
  • Crystallography, X-Ray
  • Histones / chemistry*
  • Histones / genetics
  • Humans
  • Immunoglobulin Fab Fragments / chemistry*
  • Immunoglobulin Fab Fragments / genetics
  • Methylation
  • Protein Structure, Quaternary


  • Antibodies, Monoclonal
  • Antigens
  • Histones
  • Immunoglobulin Fab Fragments

Associated data

  • PDB/4YHP
  • PDB/4YHY
  • PDB/4YHZ