Affinity chromatography: A versatile technique for antibody purification

Methods. 2017 Mar 1:116:84-94. doi: 10.1016/j.ymeth.2016.12.010. Epub 2016 Dec 22.

Abstract

Antibodies continue to be extremely utilized entities in myriad applications including basic research, imaging, targeted delivery, chromatography, diagnostics, and therapeutics. At production stage, antibodies are generally present in complex matrices and most of their intended applications necessitate purification. Antibody purification has always been a major bottleneck in downstream processing of antibodies, due to the need of high quality products and associated high costs. Over the years, extensive research has focused on finding better purification methodologies to overcome this holdup. Among a plethora of different techniques, affinity chromatography is one of the most selective, rapid and easy method for antibody purification. This review aims to provide a detailed overview on affinity chromatography and the components involved in purification. An array of support matrices along with various classes of affinity ligands detailing their underlying working principles, together with the advantages and limitations of each system in purifying different types of antibodies, accompanying recent developments and important practical methodological considerations to optimize purification procedure are discussed.

Keywords: Affinity chromatography; Affinity ligands; Affinity matrices; Antibody; Antibody purification.

Publication types

  • Review

MeSH terms

  • Animals
  • Antibodies / chemistry
  • Antibodies / isolation & purification*
  • Chromatography, Affinity / instrumentation
  • Chromatography, Affinity / methods*
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Ligands
  • Protein Binding
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Static Electricity

Substances

  • Antibodies
  • Ligands
  • Recombinant Proteins