Recovery and purification of recombinant protein from transgenic corn by aqueous two-phase partitioning

摘要

Com has been used as an expression host for several recombinant proteins with potential for large-scale production. Cost-effective downstream initial recovery, separation and concentration remain a challenge. Aqueous two-phase (ATP) partitioning has been used to recover and concentrate proteins from fermentation broths and offers advantages for integration of those steps with biomass removal.rnAn integrated procedure, which combines extraction and partitioning into one step, was compared to partitioning of separately prepared clarified extracts (two-step procedures). The results of the integrated procedure showed less protein extracted and lower protein partitioning to the upper phase than in the two-step procedures. A model protein, lysozyme, was added to the integrated extract to assess separation selectivity of this method for a protein that had previously been enriched by the two-step procedures. Phase ratio and NaCl addition were manipulated to optimize separation by ATP partitioning. NaCl addition and lowering the phase ratio enhanced the separation of lysozyme from com proteins.