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

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• 作者：Maria B. Dainiak ; Igor Yu. Galaev and Bo Mattiasson
• 刊名：Journal of Chromatography A
• 出版年：2006
• 出版时间：11 August 2006
• 年：2006
• 卷：1123
• 期：2
• 页码：145-150
• 全文大小：1846 K

文摘

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 × 7.1 mm Ø) with immobilized concanavalin A was used. Chromatographic columns (113 mm × 7.1 mm Ø) 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 b1;-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.