CaptureSelect FcXP affinity medium exhibits strong aggregate separation capability

Protein Expr Purif. 2024 Aug:220:106503. doi: 10.1016/j.pep.2024.106503. Epub 2024 May 15.

Abstract

Protein A affinity chromatography has been widely used for initial product capture in recombinant antibody/Fc-fusion purification. However, in general Protein A lacks the capability of separating aggregates (unless the aggregates are too large to enter the pores of resin beads or have their Protein A binding sites buried, in which case the aggregates do not bind). In the current work, we demonstrated that CaptureSelect FcXP affinity medium exhibited strong aggregate separation capability and effectively removed aggregates under pH or conductivity gradient elution in two bispecific antibody (bsAb) cases. For these two cases, aggregate contents were reduced from >16% and >22% (in the feed) to <1% and <5% (in the eluate) for the first and second bsAbs, respectively. While more case studies are required to further demonstrate FcXP's superiority in aggregate removal, findings from the current study suggest that FcXP can potentially be a better alternative than Protein A for product capture in cases where aggregate content is high.

Keywords: Aggregate; Bispecific antibody (bsAb); CaptureSelect FcXP; Conductivity gradient; Protein A; pH gradient.

MeSH terms

  • Antibodies, Bispecific* / chemistry
  • Antibodies, Bispecific* / isolation & purification
  • Chromatography, Affinity* / methods
  • Humans
  • Immunoglobulin Fc Fragments / chemistry
  • Immunoglobulin Fc Fragments / isolation & purification
  • Protein Aggregates
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Staphylococcal Protein A* / chemistry

Substances

  • Antibodies, Bispecific
  • Staphylococcal Protein A
  • Recombinant Fusion Proteins
  • Protein Aggregates
  • Immunoglobulin Fc Fragments