Deamidation and isomerization liability analysis of 131 clinical-stage antibodies

MAbs. 2019 Jan;11(1):45-57. doi: 10.1080/19420862.2018.1548233. Epub 2018 Dec 10.


Contemporary in vivo and in vitro discovery platform technologies greatly increase the odds of identifying high-affinity monoclonal antibodies (mAbs) towards essentially any desired biologically relevant epitope. Lagging discovery throughput is the ability to select for highly developable mAbs with drug-like properties early in the process. Upstream consideration of developability metrics should reduce the frequency of failures in later development stages. As the field moves towards incorporating biophysical screening assays in parallel to discovery processes, similar approaches should also be used to ensure robust chemical stability. Optimization of chemical stability in the early stages of discovery has the potential to reduce complications in formulation development and improve the potential for successful liquid formulations. However, at present, our knowledge of the chemical stability characteristics of clinical-stage therapeutic mAbs is fragmented and lacks comprehensive comparative assessment. To address this knowledge gap, we produced 131 mAbs with amino acid sequences corresponding to the variable regions of clinical-stage mAbs, subjected these to low and high pH stresses and identified the resulting modifications at amino acid-level resolution via tryptic peptide mapping. Among this large set of mAbs, relatively high frequencies of asparagine deamidation events were observed in CDRs H2 and L1, while CDRs H3, H2 and L1 contained relatively high frequencies of instances of aspartate isomerization.

Keywords: antibody; asparagine deamidation; aspartic acid isomerization; chemical liability; developability; forced degradation; low and high pH stress; tryptic peptide mapping.

MeSH terms

  • Antibodies, Monoclonal / chemistry*
  • Complementarity Determining Regions / chemistry
  • Drug Discovery / methods*
  • Humans
  • Isomerism
  • Protein Stability


  • Antibodies, Monoclonal
  • Complementarity Determining Regions