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

Kumar, Ashok LU ; Rodriguez-Caballero, A; Plieva, Fatima LU ; Galaev, Igor LU ; Nandakumar, K S; Kamihira, M; Holmdahl, Rikard LU ; Orfao, A and Mattiasson, Bo LU (2005) In Journal of Molecular Recognition 18(1). p.84-93
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Molecular Recognition
volume
18
issue
1
pages
84 - 93
publisher
John Wiley & Sons
external identifiers
  • wos:000227612900006
  • scopus:11244305393
ISSN
1099-1352
DOI
10.1002/jmr.693
language
English
LU publication?
yes
id
c96c8be8-979c-4685-b0d9-d1da4d39ef97 (old id 155189)
date added to LUP
2007-07-02 12:04:23
date last changed
2017-01-15 03:29:40
@article{c96c8be8-979c-4685-b0d9-d1da4d39ef97,
  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.},
  author       = {Kumar, Ashok and Rodriguez-Caballero, A and Plieva, Fatima and Galaev, Igor and Nandakumar, K S and Kamihira, M and Holmdahl, Rikard and Orfao, A and Mattiasson, Bo},
  issn         = {1099-1352},
  language     = {eng},
  number       = {1},
  pages        = {84--93},
  publisher    = {John Wiley & Sons},
  series       = {Journal of Molecular Recognition},
  title        = {Affinity binding of cells to cryogel adsorbents with immobilized specific ligands: effect of ligand coupling and matrix architecture},
  url          = {http://dx.doi.org/10.1002/jmr.693},
  volume       = {18},
  year         = {2005},
}