title: Effects of peptide density and elution pH on affinity chromatographic purification of human immunoglobulins A and M.
abstract: A family of linear hexamer peptide ligands HWRGWV, HYFKFD and HFRRHL, initially identified for their affinity to the Fc portion of human immunoglobulin G (hIgG), also have potential for use in the purification of human immunoglobulins A (hIgA) and M (hIgM). HWRGWV demonstrated the strongest binding affinity to hIgM, followed by hIgA and hIgG respectively. The effects of N-terminal acetylation of the peptide, as well as elution buffer pH, on the chromatographic elution of human IgG, IgA and IgM from HWRGWV resins at various peptide densities (0.04-0.55 meq/g) were investigated. Over 80% recovery and 90% purity were achieved for human IgG and IgA isolation from complete minimum essential medium (cMEM) using HWRGWV resin at optimum peptide densities. For human IgM, 75.7% recovery and 86.0% purity were achieved by using HWRGWV at a low peptide density of 0.04 meq/g. Although HYFKFD and HFRRHL exhibited their ability for isolation of human IgG, IgA and IgM from cMEM as well, HWRGWV is the best option among them for large-scale purification of human IgG, IgA and IgM based on conditions tested.
authors: Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695, USA.
authors: Zhuo
authors: Z
authors: Liu
authors: Patrick V
authors: PV
authors: Gurgel
authors: Ruben G
authors: RG
authors: Carbonell
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007070:Immunoglobulin A / Q000737:chemistry / Q000302:isolation & purification*; D007075:Immunoglobulin M / Q000737:chemistry / Q000302:isolation & purification*; D007163:Immunosorbent Techniques; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry*; D011487:Protein Conformation; D016415:Sequence Alignment
chemical_list: D007070:Immunoglobulin A; D007075:Immunoglobulin M; D009842:Oligopeptides