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

Ashok Kumar, A Rodriguez-Caballero, Fatima Plieva, Igor Galaev, K S Nandakumar, M Kamihira, Rikard Holmdahl, A Orfao, Bo Mattiasson

Research output: Contribution to journalArticlepeer-review

72 Citations (SciVal)

Abstract

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.
Original languageEnglish
Pages (from-to)84-93
JournalJournal of Molecular Recognition
Volume18
Issue number1
DOIs
Publication statusPublished - 2005

Bibliographical note

The information about affiliations in this record was updated in December 2015.
The record was previously connected to the following departments: Medical Inflammation Research (013212019), Biotechnology (LTH) (011001037)

Subject classification (UKÄ)

  • Immunology in the medical area

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