Improving the extraction and purification of immunoglobulin G by the use of ionic liquids as adjuvants in aqueous biphasic systems

J Biotechnol. 2016 Oct 20:236:166-175. doi: 10.1016/j.jbiotec.2016.08.015. Epub 2016 Aug 25.

Abstract

Immunoglobulins G (IgG) could become widespread biopharmaceuticals if cost-efficient processes for their extraction and purification are available. In this work, aqueous biphasic systems (ABS) composed of polyethylene glycols and a buffered salt, and with ionic liquids (ILs) as adjuvants, have been studied as alternative extraction and purification platforms of IgG from a rabbit serum source. Eleven ILs were investigated to provide insights on the chemical features which maximize the IgG partitioning. It is shown that in polymer-salt systems pure IgG preferentially partitions to the polymer-rich phase; yet, the complete extraction was never attained. Remarkably, after the addition of 5wt% of adequate ILs to polymer-salt ABS, the complete extraction of pure IgG in a single-step was accomplished. The best systems and conditions were then applied to the extraction and purification of IgG directly from rabbit serum samples. The complete extraction of IgG in a single-step was maintained while its purity in the polymer-rich phase was enhanced by ca. 37% as compared to the IL-free ABS. The antibody stability was also evaluated revealing that appropriate ILs are able to maintain the IgG stability and can be used as phase-forming components of ABS when envisaging the purification of high-cost biopharmaceuticals.

Keywords: Adjuvants; Antibody; Aqueous biphasic system; Extraction; Ionic liquids; Purification.

MeSH terms

  • Animals
  • Hydrogen-Ion Concentration
  • Imidazoles / chemistry
  • Immunoglobulin G / blood
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / isolation & purification*
  • Ionic Liquids / chemistry*
  • Liquid-Liquid Extraction / methods*
  • Polyethylene Glycols / chemistry*
  • Protein Stability
  • Rabbits

Substances

  • Imidazoles
  • Immunoglobulin G
  • Ionic Liquids
  • Polyethylene Glycols