Detachment strategies for affinity-adsorbed cells

X Cao, R Eisenthal, J Hubble

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Target cells adsorbed in affinity sepn. processes need to be effectively recovered from the adsorption matrix once contaminating cell types have been removed. Although tangential flow can be used to generate detachment forces, the resultant diln. is excessive. In this study, three alternative detachment protocols have been assessed using a membrane-based adsorption surface. Back flushing buffer through the membrane to generate greater detachment forces was studied. However, at the max. back flush velocity obtainable less than 50% of attached cells were removed and recovered cell concns. were low. Controlled transmembrane delivery of hydrochloric acid as a non-specific eluent was also investigated. While this allowed complete recovery of cells the effective acid concn. was excessive. The upstream diln. required to maintain cell viability again leads to considerable diln.Injection of air bubbles into the tangential flow stream to generate elevated detachment forces was found to be the most effective approach. Detachment was found to be a function of both no. and size of bubbles. Under optimal conditions, 80% of attached cells can be removed to give a detached cell concn. of 108 cells ml-1. This leads to an essentially reagentless detachment protocol allowing the development of an isocratic affinity cell separator where cells can be attached, washed, and detached in the same buffer. [on SciFinder (R)]
Original languageEnglish
Pages (from-to)153-160
Number of pages8
JournalEnzyme Microbial Technology
Volume31
Issue number1-2
Publication statusPublished - 2002

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Buffers
Membranes
Adsorption
Hydrochloric Acid
Separators
Cells
Recovery
Acids
Air
Cell Survival
Injections

Keywords

  • Shear
  • BUU (Biological use
  • detachment wine yeast affinity cell separator
  • affinity cell adsorption
  • Cell
  • Desorption (of cells
  • Separators (sorters
  • unclassified)
  • BIOL (Biological study)
  • Immobilization (microbial cell
  • Saccharomyces cerevisiae
  • Polyamides Role
  • Filtration (tangential-flow filtration
  • Adsorption
  • detachment strategies for affinity-adsorbed cells)
  • Bubbles
  • Immobilization
  • Wine yeast (detachment strategies for affinity-adsorbed cells)
  • USES (Uses) (detachment strategies for affinity-adsorbed cells)

Cite this

Cao, X., Eisenthal, R., & Hubble, J. (2002). Detachment strategies for affinity-adsorbed cells. Enzyme Microbial Technology, 31(1-2), 153-160.

Detachment strategies for affinity-adsorbed cells. / Cao, X; Eisenthal, R; Hubble, J.

In: Enzyme Microbial Technology, Vol. 31, No. 1-2, 2002, p. 153-160.

Research output: Contribution to journalArticle

Cao, X, Eisenthal, R & Hubble, J 2002, 'Detachment strategies for affinity-adsorbed cells', Enzyme Microbial Technology, vol. 31, no. 1-2, pp. 153-160.
Cao X, Eisenthal R, Hubble J. Detachment strategies for affinity-adsorbed cells. Enzyme Microbial Technology. 2002;31(1-2):153-160.
Cao, X ; Eisenthal, R ; Hubble, J. / Detachment strategies for affinity-adsorbed cells. In: Enzyme Microbial Technology. 2002 ; Vol. 31, No. 1-2. pp. 153-160.
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