Acidic pH or salt treatment can convert soluble antibody aggregates in culture harvest into monomers and improve Protein A chromatography step yield

Protein Expr Purif. 2024 Mar:215:106391. doi: 10.1016/j.pep.2023.106391. Epub 2023 Nov 7.


While purifying a regular monospecific antibody, we found that the Protein A step yield was much lower than expected. Further studies revealed that the antibody formed large-size aggregates that did not bind to the Protein A resin, hence leading to dropped recovery. In an attempt to solve this low yield issue, we found that mildly acidic pH or ammonium sulfate treatment can partially convert the aggregates into monomers. In addition, when acidic pH treated culture harvest was processed by Protein A chromatography, the yield was restored to the normal range, suggesting that the monomers recovered from aggregates regained Protein A binding capability. Thus, low pH treatment of culture harvest can be potentially used as a general approach for improving Protein A step yield in cases where non-binding antibody aggregates are formed through noncovalent interactions.

Keywords: Acidic pH; Aggregate; Antibody; Monomer; Protein A chromatography; Salt.

MeSH terms

  • Antibodies, Monoclonal* / chemistry
  • Chromatography
  • Hydrogen-Ion Concentration
  • Staphylococcal Protein A* / chemistry


  • Antibodies, Monoclonal
  • Staphylococcal Protein A