Effect of pre-filtration on selective isolation of tat protein by affinity membrane separation: Analysis of flux, separation efficiency, and processing time

Saurav Datta, D. Bhattacharyya, P. D. Ray, A. Nath, M. Toborek

Research output: Contribution to journalArticle

Abstract

The isolation and purification of Tat protein from bacterial lysate using avidin-biotin interaction in microfiltration membranes have been reported in the literature. To increase the efficacy of the technique, improvements in flux, Tat separation efficiency, and processing time are essential. In the current research work a pre-filtration step was introduced to remove unwanted high molecular weight proteins and other impurities from feed prior to affinity membrane separation. Significant enhancement in flux and separation efficiency of Tat was observed. Processing time was also reduced significantly. For example, with UF pretreatment step the total Tat recovery was around four times higher (with processing time 25% lower) than that observed with the untreated feed. The quality of purified Tat was analyzed by SDS-PAGE, Western Blot, and biotin analysis. Flux behavior in affinity separation was described by model equations.

Original languageEnglish (US)
Pages (from-to)2451-2471
Number of pages21
JournalSeparation Science and Technology
Volume42
Issue number11
DOIs
StatePublished - Aug 1 2007
Externally publishedYes

Keywords

  • Affinity separation
  • Avidin
  • Biotin
  • Flux
  • Processing time
  • Separation efficiency
  • Tat

ASJC Scopus subject areas

  • Chemistry(all)
  • Chemical Engineering(all)
  • Process Chemistry and Technology
  • Filtration and Separation

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