Purification of human immunoglobulin G via Fc-specific small peptide ligand affinity chromatography.
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.
Yang
Haiou
H
Department of Chemical and Biomolecular Engineering, North Carolina State University, 1017 Main Campus Drive, Centennial Campus, Partner's Building I, Suite 3200, Box 7006, Raleigh, NC 27695-7565, USA.
Gurgel
Patrick V
PV
Carbonell
Ruben G
RG
D000327:Adsorption; D002846:Chromatography, Affinity / Q000379:methods*; D003470:Culture Media; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism*; D007141:Immunoglobulin Fc Fragments / Q000378:metabolism*; D007074:Immunoglobulin G / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D013696:Temperature
D003470:Culture Media; D055767:Immobilized Proteins; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D008024:Ligands; D010455:Peptides