Purification of human immunoglobulin G via Fc-specific small peptide ligand affinity chromatography

• Published in: Journal of Chromatography A Vol. 1216; no. 6; pp. 910 - 918
• Main Authors: Yang, Haiou, Gurgel, Patrick V., Carbonell, Ruben G.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 06-02-2009; Amsterdam; New York: Elsevier; Elsevier
• Subjects: Adsorption; Affinity chromatography; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chromatography, Affinity - methods; Culture Media; Fc fragment; Fundamental and applied biological sciences. Psychology; General pharmacology; Glycoproteins; Hexamer peptide ligand; Human immunoglobulin G; Humans; Immobilized Proteins - chemistry; Immobilized Proteins - metabolism; Immunoglobulin Fc Fragments - metabolism; Immunoglobulin G - isolation & purification; Immunoglobulin G - metabolism; Kinetics; Ligand density; Ligands; Medical sciences; Peptides - chemistry; Peptides - metabolism; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Proteins; Temperature; Human immunoglobulin G; Fc fragment; Hexamer peptide ligand; Affinity chromatography; Ligand density; Human; Purification; Peptides; Antibody; Glycoprotein; Temperature effect; IgG; Coverage rate; Operating conditions; Affinity adsorbent; Separation method; Fc-Fragment; Isolation
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2008.12.004

Chromatographic resins of a family of linear Fc-binding hexamer peptides (HWRGWV, HYFKFD, and HFRRHL) exhibited the ability to selectively adsorb and isolate human IgG (hIgG) from complete mammalian cell culture medium (cMEM). Among them, the HWRGWV resin with a peptide density of 0.08
mequiv./g of resin was able to purify hIgG from cMEM with both purity and yield as high as 95%, comparable to Protein A and A2P agarose gels. The influences of N-terminal acetylation of the HWRGWV resin, ligand density on the resin, initial hIgG concentration, and temperature on IgG isolation were also investigated. The results indicate that these small peptide ligands, especially HWRGWV, offer a potential alternative to the use of Protein A or Protein G for large scale affinity chromatography.