Affinity binding of cells to cryogel adsorbents with immobilized specific ligands: effect of ligand coupling and matrix architecture.
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.
Kumar
Ashok
A
Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, SE-22100 Lund, Sweden.
Rodríguez-Caballero
Arancha
A
Plieva
Fatima M
FM
Galaev
Igor Yu
IY
Nandakumar
Kutty Selva
KS
Kamihira
Masamichi
M
Holmdahl
Rikard
R
Orfao
Alberto
A
Mattiasson
Bo
B
D018941:Antigens, CD19 / Q000276:immunology; D018952:Antigens, CD34 / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology*; D002469:Cell Separation / Q000379:methods*; D006801:Humans; D007164:Immunosorbents / Q000737:chemistry*; D007951:Leukemia, Myeloid / Q000276:immunology; D008024:Ligands; D008863:Microspheres; D011142:Polyvinyl Alcohol / Q000737:chemistry*; D013205:Staphylococcal Protein A / Q000737:chemistry* / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology
D018941:Antigens, CD19; D018952:Antigens, CD34; D007164:Immunosorbents; D008024:Ligands; D013205:Staphylococcal Protein A; D011142:Polyvinyl Alcohol