Affinity fractionation of lymphocytes using a monolithic cryogel.
(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:
https://lup.lub.lu.se/record/129162
- author
- Kumar, Ashok LU ; Plieva, Fatima LU ; Galaev, Igor LU and Mattiasson, Bo LU
- organization
- publishing date
- 2003
- 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
- 2016-04-01 16:25:09
- date last changed
- 2023-08-15 13:30:56
@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}},
keywords = {{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}},
doi = {{10.1016/j.jim.2003.09.017}},
volume = {{283}},
year = {{2003}},
}