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Affinity cryogel monoliths for screening for optimal separation conditions and chromatographic separation of cells

Dainiak, Maria LU ; Galaev, Igor LU and Mattiasson, Bo LU (2006) In Journal of Chromatography A 1123(2). p.145-150
Abstract
Suitable conditions for separating cells using a chromatographic procedure were evaluated in parallel chromatography on minicolumns. A 96-hole minicolumn plate filled with cryogel monoliths (18.8 mm x 7.1 mm O) with immobilized concanavalin A was used. Chromatographic columns (113 mm x 7.1 mm O) were used for chromatographic resolution of a mixture of Saccharomyces cerevisiae and Escherichia coli cells. Separation of a cell mixture containing equal amounts of cells of both types performed in a column format under the determined optimal conditions, resulted in a quantitative capture of applied S. cerevisiae cells, while E. coli passed through the column. Bound S. cerevisiae cells were released by flow-induced detachment and by compression... (More)
Suitable conditions for separating cells using a chromatographic procedure were evaluated in parallel chromatography on minicolumns. A 96-hole minicolumn plate filled with cryogel monoliths (18.8 mm x 7.1 mm O) with immobilized concanavalin A was used. Chromatographic columns (113 mm x 7.1 mm O) were used for chromatographic resolution of a mixture of Saccharomyces cerevisiae and Escherichia coli cells. Separation of a cell mixture containing equal amounts of cells of both types performed in a column format under the determined optimal conditions, resulted in a quantitative capture of applied S. cerevisiae cells, while E. coli passed through the column. Bound S. cerevisiae cells were released by flow-induced detachment and by compression of the adsorbent in the presence of 0.3 M methyl U-D-manno-pyranoside. The flowthrough and the eluted fractions were analyzed by plate counting and by registering metabolic activity of S. cerevisiae cells in the eluted fractions after capturing on ConA-cryogel monoliths in a 96-minicolumn plate format. The flowthrough fraction contained E. coli cells with nearly 100% purity, whereas the fraction eluted by compression of the adsorbent contained viable S. cerevisiae cells with 95% purity. Thus, an efficient chromatographic separation of cells was achieved using affinity cryogel column. (Less)
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author
; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
screening, column format, format, 96-minicolumn plate, elastic, ConA-cryogel monoliths, macroporous, cell chromatography, cell detachment, compression of the adsorbent
in
Journal of Chromatography A
volume
1123
issue
2
pages
145 - 150
publisher
Elsevier
external identifiers
  • wos:000239741400002
  • pmid:16846611
  • scopus:33746337923
ISSN
0021-9673
DOI
10.1016/j.chroma.2006.05.089
language
English
LU publication?
yes
id
e111c585-66f8-43ac-86f7-2fa592aaf2ad (old id 397383)
date added to LUP
2016-04-01 16:59:42
date last changed
2021-08-25 04:02:56
@article{e111c585-66f8-43ac-86f7-2fa592aaf2ad,
  abstract     = {Suitable conditions for separating cells using a chromatographic procedure were evaluated in parallel chromatography on minicolumns. A 96-hole minicolumn plate filled with cryogel monoliths (18.8 mm x 7.1 mm O) with immobilized concanavalin A was used. Chromatographic columns (113 mm x 7.1 mm O) were used for chromatographic resolution of a mixture of Saccharomyces cerevisiae and Escherichia coli cells. Separation of a cell mixture containing equal amounts of cells of both types performed in a column format under the determined optimal conditions, resulted in a quantitative capture of applied S. cerevisiae cells, while E. coli passed through the column. Bound S. cerevisiae cells were released by flow-induced detachment and by compression of the adsorbent in the presence of 0.3 M methyl U-D-manno-pyranoside. The flowthrough and the eluted fractions were analyzed by plate counting and by registering metabolic activity of S. cerevisiae cells in the eluted fractions after capturing on ConA-cryogel monoliths in a 96-minicolumn plate format. The flowthrough fraction contained E. coli cells with nearly 100% purity, whereas the fraction eluted by compression of the adsorbent contained viable S. cerevisiae cells with 95% purity. Thus, an efficient chromatographic separation of cells was achieved using affinity cryogel column.},
  author       = {Dainiak, Maria and Galaev, Igor and Mattiasson, Bo},
  issn         = {0021-9673},
  language     = {eng},
  number       = {2},
  pages        = {145--150},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography A},
  title        = {Affinity cryogel monoliths for screening for optimal separation conditions and chromatographic separation of cells},
  url          = {http://dx.doi.org/10.1016/j.chroma.2006.05.089},
  doi          = {10.1016/j.chroma.2006.05.089},
  volume       = {1123},
  year         = {2006},
}