High throughput purification of monoclonal recombinant antibodies using a Protein-A coated membrane plate system

N Biotechnol. 2023 Nov 25:77:111-119. doi: 10.1016/j.nbt.2023.08.003. Epub 2023 Aug 28.

Abstract

The therapeutic use of monoclonal antibodies (mAbs) ranges from cancer treatment to immune-mediated conditions, covering infectious and cardiovascular disorders, among others. The development of improved methods for therapeutic antibody discovery has accelerated the identification of numerous mAbs: a discovery campaign can be deeply mined, resulting in hundreds, even thousands, of potential antibody leads for a given target of interest. High throughput mAb expression and purification methods are required for the rapid validation of those leads. In this work, we describe the implementation of a Protein-A coated membrane plate system, the Purexa™ AHT membrane plate, for robust preparative purification of hundreds of recombinant mAbs, without the need for automation. The high efficiency (>80%) recovery generated sufficient mAb for downstream screening analyses such as ELISA and surface plasmon resonance (SPR). This new system allows the functional validation of hundreds of lead antibodies from discovery campaigns in a timely manner regardless of operational size.

Keywords: Affinity purification; Antibodies; High throughput; Membrane plate; Protein A.

MeSH terms

  • Antibodies, Monoclonal*
  • Enzyme-Linked Immunosorbent Assay
  • Recombinant Proteins
  • Staphylococcal Protein A*
  • Surface Plasmon Resonance

Substances

  • Antibodies, Monoclonal
  • Recombinant Proteins
  • Staphylococcal Protein A