Advanced

Affinity fractionation of lymphocytes using a monolithic cryogel.

Kumar, Ashok LU ; Plieva, Fatima LU ; Galaev, Igor LU and Mattiasson, Bo LU (2003) In Journal of Immunological Methods 283(1-2). p.185-194
Abstract
A new type of continuous, supermacroporous, monolithic, cryogel affinity adsorbent was developed, allowing specific fractionation and separation of human peripheral blood lymphocytes in a chromatographic format. The affinity adsorbent was used to design a novel cell separation strategy, which was based on the interaction of protein A from Staphylococcus aureus with cells bearing IgG antibodies on the surface. After treating lymphocytes with goat anti-human IgG(H+L), the IgG-positive B-lymphocytes were efficiently separated from T-lymphocytes. Protein A covalently coupled to epoxy activated dimethylacrylamide (DMAA) cryogel matrix specifically bound IgG-bearing B-lymphocytes through the Fc region, while non-bound T-lymphocytes passed... (More)
A new type of continuous, supermacroporous, monolithic, cryogel affinity adsorbent was developed, allowing specific fractionation and separation of human peripheral blood lymphocytes in a chromatographic format. The affinity adsorbent was used to design a novel cell separation strategy, which was based on the interaction of protein A from Staphylococcus aureus with cells bearing IgG antibodies on the surface. After treating lymphocytes with goat anti-human IgG(H+L), the IgG-positive B-lymphocytes were efficiently separated from T-lymphocytes. Protein A covalently coupled to epoxy activated dimethylacrylamide (DMAA) cryogel matrix specifically bound IgG-bearing B-lymphocytes through the Fc region, while non-bound T-lymphocytes passed through the column. More than 90% of the B-lymphocytes were retained in the column while the cells in the breakthrough fraction were enriched in T-lymphocytes (81%). The viability of the T-lymphocytes isolated was greater than 90%. The bound lymphocytes released by human or dog IgG recovered 60–70% of the B-cells without significantly impairing the cell viability. The technique can be applied in general to cell separation systems where IgG antibodies against specific cell surface markers are available. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Cell separation, Supermacroporous cryogels, Lymphocyte fractionation, Monolithic adsorbent, Protein-A cell affinity
in
Journal of Immunological Methods
volume
283
issue
1-2
pages
185 - 194
publisher
Elsevier
external identifiers
  • pmid:14659910
  • wos:000187349900017
  • scopus:0344196813
ISSN
1872-7905
DOI
10.1016/j.jim.2003.09.017
language
English
LU publication?
yes
id
507f99d6-4062-4fb3-87ee-627ac12127b2 (old id 129162)
date added to LUP
2007-07-02 12:06:04
date last changed
2018-10-14 04:09:56
@article{507f99d6-4062-4fb3-87ee-627ac12127b2,
  abstract     = {A new type of continuous, supermacroporous, monolithic, cryogel affinity adsorbent was developed, allowing specific fractionation and separation of human peripheral blood lymphocytes in a chromatographic format. The affinity adsorbent was used to design a novel cell separation strategy, which was based on the interaction of protein A from Staphylococcus aureus with cells bearing IgG antibodies on the surface. After treating lymphocytes with goat anti-human IgG(H+L), the IgG-positive B-lymphocytes were efficiently separated from T-lymphocytes. Protein A covalently coupled to epoxy activated dimethylacrylamide (DMAA) cryogel matrix specifically bound IgG-bearing B-lymphocytes through the Fc region, while non-bound T-lymphocytes passed through the column. More than 90% of the B-lymphocytes were retained in the column while the cells in the breakthrough fraction were enriched in T-lymphocytes (81%). The viability of the T-lymphocytes isolated was greater than 90%. The bound lymphocytes released by human or dog IgG recovered 60–70% of the B-cells without significantly impairing the cell viability. The technique can be applied in general to cell separation systems where IgG antibodies against specific cell surface markers are available.},
  author       = {Kumar, Ashok and Plieva, Fatima and Galaev, Igor and Mattiasson, Bo},
  issn         = {1872-7905},
  keyword      = {Cell separation,Supermacroporous cryogels,Lymphocyte fractionation,Monolithic adsorbent,Protein-A cell affinity},
  language     = {eng},
  number       = {1-2},
  pages        = {185--194},
  publisher    = {Elsevier},
  series       = {Journal of Immunological Methods},
  title        = {Affinity fractionation of lymphocytes using a monolithic cryogel.},
  url          = {http://dx.doi.org/10.1016/j.jim.2003.09.017},
  volume       = {283},
  year         = {2003},
}