Antibody Purification Using Affinity Chromatography

Methods Mol Biol. 2022:2466:3-22. doi: 10.1007/978-1-0716-2176-9_1.

Abstract

Antibodies are an integral part of many biological assays and biotherapeutics. However, the sources from which antibodies are derived frequently contain other contaminants which may interfere with assays or cause adverse reactions if administered in vivo. Therefore, a means of isolating these antibodies from their source at high levels of purity is critical. Affinity chromatography is currently one of the most widely applied methods for the purification of antibodies. This method relies on specific and reversible, interactions between antibody structures, or recombinant tags fused to these structures, and ligands immobilized on solid support matrices, generally within a column. Herein, common chromatographic methods applied to antibody purification are described. These include the purification of IgG, and its recombinant forms, through protein A, protein G and immobilized metal affinity chromatography.

Keywords: Affinity purification; Antibodies; Chromatography; Methods; Recombinant proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies* / chemistry
  • Chromatography, Affinity / methods
  • Ligands
  • Staphylococcal Protein A* / chemistry

Substances

  • Antibodies
  • Ligands
  • Staphylococcal Protein A