Performance comparison of protein A affinity resins for the purification of monoclonal antibodies

J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Mar 15;848(1):97-107. doi: 10.1016/j.jchromb.2006.04.050. Epub 2006 Jun 12.

Abstract

During the selection of protein A affinity resin for the purification of monoclonal antibodies, dynamic binding capacity (Q(dyn10%)), volumetric production rate (Pr(vol)) and 'process robustness' are essential parameters to be evaluated. In this article, empirical mathematical models describe these parameters as a function of antibody concentration in load (C0), load flow rate (u(load)) and bed height (L). These models allow us to select optimal process conditions for each of the evaluated protein A affinity resins. C0, u(load) and L largely affect dynamic binding capacity (Q(dyn10%)) and volumetric production rate (Pr(vol)). Maximum Q(dyn10%) is generally obtained at high C0 and at low u(load). Maximum Pr(vol) is obtained at high C0 and at lowest L, run at high u(load). All evaluated resins have a relatively high robustness against variations in C0. |DeltaQ(dyn10%)/deltaC0| ranges from 0.0 to 7.8. It is clear that Q(dyn10%), Pr(vol) and 'process robustness' cannot be maximized all at the same time. Furthermore, some other aspects like IgG recovery, protein A leaching, easiness to pack, easiness to clean, number of re-uses and cost of production might be important to be taken into the equation. Certain evaluation parameters may be more important than others, depending on the specific situation. Therefore, a case-by-case evaluation is recommended.

MeSH terms

  • Algorithms
  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / isolation & purification*
  • Chromatography, Affinity / methods*
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Hybridomas
  • Immunoglobulin G / biosynthesis
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / isolation & purification
  • Mice
  • Reproducibility of Results
  • Staphylococcal Protein A / chemistry*

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin G
  • Staphylococcal Protein A