Impact of operating variables on the expanded bed adsorption of Saccharomyces cerevisiae cells using a concanavalin A derivatized perfluorocarbon

Abstract

The use of fluidizable affinity adsorbents for the adsorption of cells in expanded mode is investigated. Affinity adsorbents have been synthesized by immobilizing the lectin Concanavalin A onto the surface of triazine-activated perfluorocarbon-solids. The adsorbents were found to adsorb Saccharomyces cerevisiae cells from solution with adsorption capacities of up to 6.8 x 10(9) cells mL(-1). Adsorption kinetics were rapid with a time constant of </=8 min. The adsorbed cells could be eluted using 500 mM methyl alpha,D-mannopyranoside, although the kinetics of release were slowed by the multipoint nature of the interaction. The dynamic capacity of the Con A PVA FEP in expanded mode was up to 4.5 x 10(9) cells mL(-1). The operating parameters of bed height, application flow rate, and adsorbent size distribution were investigated for any potential improvements in throughput, which may improve utility for more fragile cells. A decrease in settled-bed height from 20 to 5 cm resulted in a decrease in dynamic capacity of 27% from 4.5 to 3.3 x 10(9) cells mL(-1). An increase in application flow rate from 0.7 to 2.0 mL/min(-1) (resulting in an expansion increase from two- to fourfold) resulted in a 40% decrease in dynamic capacity from 4.0 to 2.4 x 10(9) cells mL(-1). An increase in the mean size distribution of the perfluorocarbon from 42 to 69 microm and therefore the flow rate needed for twofold expansion of 0.7 to 1.5 mL/min(-1) resulted in a 56% decrease in dynamic capacity from 4.0 to 1.8 x 10(9) cells mL(-1). The expanded bed, using certain combinations of the operating parameters, therefore shows significant potential for the robust, high efficiency and high capacity capture and separation of cells.

Discrimination and detection of bacteria with a label-free impedimetric biosensor based on self-assembled lectin monolayer

“…The phenomenon indicates that S. cerevisiae is captured by the Con A interface. The experimental result is agreed with previous literature reports [21,22].…”

Membrane adsorbers comprising grafted glycopolymers for targeted lectin binding

“…Many research studies have employed conA for lectin affinity chromatography. Of special interest to the authors, Clemmitt and Chase investigated the roles of operating variables and established optimal parameters for designing expanded bed columns for high capacity and high efficiency capture of specific glycosylated cells. Reichelt et al describe efforts to improve the throughput of lectin affinity chromatographic media, including the development of monolithic spin‐type columns that employ conA for enriching and purifying glycan mixtures.…”

Separation and enrichment of neural stem cells using segregation in an expanded bed

“…The expanded bed, already known for its ability to segregate particles based on size, and its various applications in adsorption, as reported by Chase and Draeger (1992), Chang and Chase (1996), Chen et al (2003), Clemmitt and Chase (2003), Lin et al (2003), Ujam et al (2003), and Abdullah and Chase (2005), is demonstrated in this paper to have a novel application in the separation and enrichment of neural stem cells.…”