title: Affinity chromatographic purification of human immunoglobulin a from Chinese hamster ovary cell culture supernatant.
abstract: Hexamer peptide ligand HWRGWV, initially screened from a solid phase combinatorial peptide library for immunoglobulins G (IgG) purification, is shown to also have potential for immunoglobulin A (IgA) purification. The determined dissociation constants for hIgA on HWRGWV resins at three different peptide densities from 0.11 to 0.55 meq/g fall in the range of 10(-6) -10(-7) M, which are somewhat lower than those for hIgG. Although relatively low dynamic binding capacity (DBC) in the range of 9.2-16.8 mg IgA/mL resin at linear flow rates from 173 to 35 cm/h were obtained for IgA compared to IgG, the DBC value of HWRGWV for IgA is much greater than current commercially available affinity ligands. Although relatively lower binding affinity to secretory IgA compared to monomeric IgA was observed, the peptide ligand resins exhibit great potential for large-scale purification of both human IgA and secretory IgA. Recoveries of 96.0% and 94.3%, and purities of 90.3% and 91.7% were achieved for human IgA and secretory IgA purification, respectively, from spiked Chinese hamster ovary cell culture supernatants without an extra afterwash step. Over 95% in purities were achieved for IgA and secretory IgA with an extra afterwash step; however, the recoveries would decrease at least 15% and 40% for IgA and secretory IgA, respectively.
authors.affiliation: Dept. of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695, USA.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Liu
authors.forename: Patrick V
authors.initials: PV
authors.lastname: Gurgel
authors.forename: Ruben G
authors.initials: RG
authors.lastname: Carbonell
mesh_terms: D000327:Adsorption; D000818:Animals; D016466:CHO Cells; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D007070:Immunoglobulin A / Q000302:isolation & purification*; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry; D013499:Surface Properties
chemical_list: D007070:Immunoglobulin A; D009842:Oligopeptides