Purification of animal immunoglobulin G (IgG) using peptoid affinity ligands

Biotechnol Prog. 2020 Jul;36(4):e2994. doi: 10.1002/btpr.2994. Epub 2020 Apr 6.


The availability of highly pure animal antibodies is critical in the production of diagnostic tools and biosensors. The peptoid PL16, previously isolated from an ensemble of peptoid variants of the IgG-binding peptide HWRGWV, was utilized in this work as affinity ligand on WorkBeads resin for the purification of immunoglobulin G (IgG) from a variety of mammalian sources and chicken immunoglobulin Y (IgY). The chromatographic protocol initially optimized for murine serum and ascites was subsequently employed for processing rabbit, goat and sheep, donkey, llama, and chicken sera. The PL16-WorkBeads resin proved able to recover all antibody targets with values of yield between 50 and 90%, and purity consistently above 90%. Notably, PL16 not only binds a broader spectrum of animal immunoglobulins than the reference ligands Protein A and G, but it also binds equally well with all their subclasses. Unlike the protein ligands, in fact, PL16 afforded excellent values of yield and purity of mammalian polyclonal IgG, namely murine (47 and 94%), rabbit (66.5 and 91.7%), caprine IgG (63 and 91-95%), donkey, and llama (93 and 97%), as well as chicken IgY (42 and 92%). Of notice, it is also the ability of PL16 to target monomeric IgG without binding aggregated IgG; when challenged with a mixture of monomeric and aggregated murine IgG, PL16 eluted <3% of fed aggregates, against 11-13% eluted by Protein A and G. Collectively, these results prove the potential of the proposed peptoid ligand for large-scale purification of animal immunoglobulins.

Keywords: IgG aggregates; affinity chromatography; animal IgG; peptoid ligands; protein purification.

MeSH terms

  • Animals
  • Chickens
  • Chromatography, Affinity*
  • Goats
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / immunology
  • Immunoglobulin G / isolation & purification*
  • Immunoglobulins / chemistry
  • Immunoglobulins / isolation & purification*
  • Ligands
  • Mice
  • Peptoids / chemistry*
  • Protein Binding / drug effects
  • Rabbits
  • Sheep


  • IgY
  • Immunoglobulin G
  • Immunoglobulins
  • Ligands
  • Peptoids