title: Comparison of antibody binding to immobilized group specific affinity ligands in high performance monolith affinity chromatography.
abstract: A novel biochromatographic principle is introduced taking the quantitative analysis of affinity interactions between antibodies and immobilized group specific ligands (protein A, G, and L) as example. The name high performance monolith affinity chromatography (HPMAC) is proposed for this technique. HPMAC uses rigid, macroporous monoliths, so-called convective interaction media (CIM)-disks, as stationary phase. An optimized procedure is described for the covalent immobilization of the group specific affinity ligands to such disks. The binding of polyclonal bovine IgG and a recombinant human antibody (type IgGl-kappa) to all affinity disks is discussed. An essential feature of HPMAC is its compatibility to unusually high mobile phase flow rates ( > 4 ml/min). Chromatographic experiments are thus completed within seconds without significant loss in binding capacity and retentive power. This makes HPMAC a promising tool for applications in fast process monitoring or screening. As an example for the former, the direct quantitative isolation of recombinant antibodies from serum-free culture supernatant is demonstrated.
authors: Center of Biotechnology, Institute of Chemical Engineering, Swiss Federal Institute of Technology, Lausanne, Switzerland.
authors: L G
authors: LG
authors: Berruex
authors: R
authors: R
authors: Freitag
authors: T B
authors: TB
authors: Tennikova
mesh_terms: D000818:Animals; D001426:Bacterial Proteins; D002417:Cattle; D002846:Chromatography, Affinity; D004268:DNA-Binding Proteins / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000378:metabolism; D013205:Staphylococcal Protein A / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C029152:G-substrate; D007074:Immunoglobulin G; C116593:L-protein, Peptococcus magnus; D009419:Nerve Tissue Proteins; D013205:Staphylococcal Protein A