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

Maria Dainiak, Igor Galaev, Bo Mattiasson

Research output: Contribution to journalArticlepeer-review

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.
Original languageEnglish
Pages (from-to)145-150
JournalJournal of Chromatography A
Volume1123
Issue number2
DOIs
Publication statusPublished - 2006

Subject classification (UKÄ)

  • Industrial Biotechnology

Free keywords

  • screening
  • column format
  • format
  • 96-minicolumn plate
  • elastic
  • ConA-cryogel monoliths
  • macroporous
  • cell chromatography
  • cell detachment
  • compression of the adsorbent

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