A comparative study of monoclonal antibodies. 1. Phase behavior and protein-protein interactions

Biotechnol Prog. Jan-Feb 2015;31(1):268-76. doi: 10.1002/btpr.2011. Epub 2014 Nov 19.


Protein phase behavior is involved in numerous aspects of downstream processing, either by design as in crystallization or precipitation processes, or as an undesired effect, such as aggregation. This work explores the phase behavior of eight monoclonal antibodies (mAbs) that exhibit liquid-liquid separation, aggregation, gelation, and crystallization. The phase behavior has been studied systematically as a function of a number of factors, including solution composition and pH, in order to explore the degree of variability among different antibodies. Comparisons of the locations of phase boundaries show consistent trends as a function of solution composition; however, changing the solution pH has different effects on each of the antibodies studied. Furthermore, the types of dense phases formed varied among the antibodies. Protein-protein interactions, as reflected by values of the osmotic second virial coefficient, are used to correlate the phase behavior. The primary findings are that values of the osmotic second virial coefficient are useful for correlating phase boundary locations, though there is appreciable variability among the antibodies in the apparent strengths of the intrinsic protein-protein attraction manifested. However, the osmotic second virial coefficient does not provide a clear basis to predict the type of dense phase likely to result under a given set of solution conditions.

Keywords: aggregation; monoclonal antibody; phase equilibrium; protein interactions; thermodynamic properties.

Publication types

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

MeSH terms

  • Ammonium Sulfate
  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / isolation & purification
  • Antibodies, Monoclonal / metabolism*
  • Humans
  • Hydrogen-Ion Concentration
  • Protein Aggregates
  • Protein Binding
  • Sodium Chloride
  • Thermodynamics


  • Antibodies, Monoclonal
  • Protein Aggregates
  • Sodium Chloride
  • Ammonium Sulfate