关键词: SDATE;Release of intracellular proteins;high speed bead mill;aqueous two-phase extraction;tumor necrosis factor

Abstract: A new technique was developed for the integrated processing of cell disruption and aqueous
two-phase extraction in a high-speed bead mill to separate intracellular proteins from
microbial cells. The process was named as simultaneous cell disruption and aqueous two-
phase extraction (SDATE). Advantages, such as, high cell disruption efficiency, biochemical
activities preserved of the proteins, cell debris elimination, and prelimiary purification
of the target protein were being claimed. When this technique was employed in isolating
recombinant Tumor Necrosis Factor (rhTNF) from E.coli, overall protein concentration and
TNF activity were found to have been increased. More than 95% of TNF was partitioned into
the top phase and all the cell debris were in the bottom phase. The partition coefficient
was greater than 3 and the TNF purification factor was greater than 6. It is shown that
less separation steps were being utilized in the new technique, meaning a reduction in
separation time and less process extractors required.

Key words: SDATE, Release of intracellular proteins, high speed bead mill, aqueous two-phase extraction, tumor necrosis factor