Affinity binding of cells to cryogel adsorbents with immobilized specific ligands: Effect of ligand coupling and matrix architecture

10 páginas, 4 figuras, 2 tablas.-- et al.The capture of human acute myeloid leukemia KG-1 cells expressing the CD34 surface antigen and the fractionation of human blood lymphocytes were evaluated on polyvinyl alcohol (PVA)-cryogel beads and dimethyl acrylamide (DMAAm) monolithic cryogel with immobilized protein A. The affinity ligand (protein A) was chemically coupled to the reactive PVA-cryogel beads and epoxy-derivatized monolithic cryogels through different immobilization techniques and the binding efficiency of the cell surface receptors specific antibody-labeled cells to the gels/beads was determined. The binding of cells to monolithic cryogel was higher (90-95%) compared with cryogel beads (76%). B-lymphocytes, which bound to the protein A-cryogel beads, were separated from T-lymphocytes with yields for the two cell types 74 and 85%, respectively. About 91% of the bound B-cells could be recovered without significantly impairing their viability. Our results show differences in the percentage of cell-binding to the immunosorbents caused by ligand density, flow shear forces and bond strength between the cells and the affinity surface once distinct chemical coupling of protein A, size of beads, sequence of antibody binding to protein A adsorbents, morphology and geometry of surface matrices were compared. Copyright 2004 John Wiley and Sons, Ltd.This work was financially supported by Protista International AB (Bjuv, Sweden), The Swedish Foundation for Strategic Research, The Swedish Foundation for International Cooperation in Research and Higher education (STINT, IG2003-2089), The Swedish Institute (Visby Program, project 2886/2002), Royal Swedish Academy of Sciences, INTAS (project 00-57) and by a grant (SAF2002-03096) from the Ministerio de Ciencia y Tecnología (Madrid, Spain). Arancha Rodríguez-Caballero is recipient of grant (13P-BPG-2003) from the Consejo Superior de Investigaciones Cientificas (CSIC-Spain).Peer Reviewe

Affinity binding of cells to cryogel adsorbents with immobilized specific ligands: Effect of ligand coupling and matrix architecture

10 páginas, 4 figuras, 2 tablas.-- et al.The capture of human acute myeloid leukemia KG-1 cells expressing the CD34 surface antigen and the fractionation of human blood lymphocytes were evaluated on polyvinyl alcohol (PVA)-cryogel beads and dimethyl acrylamide (DMAAm) monolithic cryogel with immobilized protein A. The affinity ligand (protein A) was chemically coupled to the reactive PVA-cryogel beads and epoxy-derivatized monolithic cryogels through different immobilization techniques and the binding efficiency of the cell surface receptors specific antibody-labeled cells to the gels/beads was determined. The binding of cells to monolithic cryogel was higher (90-95%) compared with cryogel beads (76%). B-lymphocytes, which bound to the protein A-cryogel beads, were separated from T-lymphocytes with yields for the two cell types 74 and 85%, respectively. About 91% of the bound B-cells could be recovered without significantly impairing their viability. Our results show differences in the percentage of cell-binding to the immunosorbents caused by ligand density, flow shear forces and bond strength between the cells and the affinity surface once distinct chemical coupling of protein A, size of beads, sequence of antibody binding to protein A adsorbents, morphology and geometry of surface matrices were compared. Copyright 2004 John Wiley and Sons, Ltd.This work was financially supported by Protista International AB (Bjuv, Sweden), The Swedish Foundation for Strategic Research, The Swedish Foundation for International Cooperation in Research and Higher education (STINT, IG2003-2089), The Swedish Institute (Visby Program, project 2886/2002), Royal Swedish Academy of Sciences, INTAS (project 00-57) and by a grant (SAF2002-03096) from the Ministerio de Ciencia y Tecnología (Madrid, Spain). Arancha Rodríguez-Caballero is recipient of grant (13P-BPG-2003) from the Consejo Superior de Investigaciones Cientificas (CSIC-Spain).Peer Reviewe

Affinity binding of cells to cryogel adsorbents with immobilized specific ligands: effect of ligand coupling and matrix architecture

The capture of human acute mycloid leukemia KG-1 cells expressing the CD34 surface antigen and the fractionation of human blood lymphocytes were evaluated on polyvinyl alcohol (PVA)-cryogel beads and dimethyl acrylamide (DMAAm) monolithic cryogel with immobilized protein A. The affinity ligand (protein A) was chemically coupled to the reactive PVA-cryogel beads and epoxy-derivatized monolithic cryogels through different immobilization techniques and the binding efficiency of the cell surface receptors specific antibody-labeled cells to the gels/beads was determined. The binding of cells to monolithic cryogel was higher (90-95%) compared with cryogel beads (76%). B-lymphocytes, which bound to the protein Acryogel beads, were separated from T-lymphocytes with yields for the two cell types 74 and 85%, respectively. About 91% of the bound B-cells could be recovered without significantly impairing their viability. Our results show differences in the percentage of cell-binding to the immunosorbents caused by ligand density, flow shear forces and bond strength between the cells and the affinity surface once distinct chemical coupling of protein A, size of beads, sequence of antibody binding to protein A adsorbents, morphology and geometry of surface matrices were compared. Copyright (c) 2004 John Wiley T Sons, Ltd