title: Initial evaluation of protein A modified capillary-channeled polymer fibers for the capture and recovery of immunoglobulin G.
abstract: A novel protein A affinity chromatography stationary phase has been developed from polypropylene capillary-channeled polymer fibers modified with a recombinant protein A ligand for the capture and recovery of immunoglobulin G (IgG) with high specificity and yield. An SPE micropipette tip format was employed so that solvent, protein, and antibody consumption was minimized. The adsorption modification of the fiber surfaces with protein A was evaluated as a function of feed concentration and volume. Optimal modification of the fiber surface with protein A yielded a 5.7 mg/mL (bed volume) ligand capacity with the modified fibers showing stability across numerous solvent environments. Performance was evaluated through exposure to human IgG and myoglobin, individually and as a mixture. Myoglobin was used as a surrogate for host cell proteins common to growth media. The efficacy of the selective binding to the ligand is demonstrated by the 2.9:1 (IgG/protein A) binding stoichiometry. Elution with 0.1 M acetic acid yielded an 89% recovery of the captured IgG based on absorption measurements of the collected eluents. Regeneration was possible with 10 mM NaOH. Protein A modified polypropylene capillary-channeled polymer fibers show promising initial results as an affinity phase for efficient capture and purification of IgG.
authors.affiliation: Department of Chemistry, Biosystems Research Complex, Clemson University, Clemson, SC, USA.
authors.forename: Abby J
authors.initials: AJ
authors.lastname: Schadock-Hewitt
authors.forename: R Kenneth
authors.initials: RK
authors.lastname: Marcus
mesh_terms: D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification*; D011108:Polymers / Q000737:chemistry*; D013205:Staphylococcal Protein A / Q000737:chemistry*
chemical_list: D007074:Immunoglobulin G; D011108:Polymers; D013205:Staphylococcal Protein A
keywords: Affinity chromatography; Capillary-channeled polymer; Fibers; Protein A; Stationary phases