期刊
JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY
卷 77, 期 9, 页码 1025-1029出版社
WILEY
DOI: 10.1002/jctb.673
关键词
aqueous two-phase systems; protein recovery; chemical extraction
In this study the use of an aqueous two-phase system (ATPS) following the direct chemical extraction of a recombinant viral coat protein, from the cytoplasm of Escherichia coli, is evaluated. The driving force is the need to establish an economically-viable process for the manufacture of a vaccine against human papilloma infection. The partition behaviour of recombinant L1 protein, the major structural protein of the virus, and DNA was investigated in a polyethylene glycol (PEG)phosphate system. An evaluation of system parameters including PEG molecular mass and the concentrations of PEG and phosphate was conducted, to estimate conditions under which the L1 protein and DNA partition to opposite phases. ATPS extraction comprising a volume ratio of 1.00, PEG 1000 (18.0%(w/w)) and phosphate (15.0%(w/w)) provided the conditions for accumulation of DNA into the bottom phase and concentration of L1 protein into the opposite phase (ie partition coefficient of DNA; In K-DNA<0.0 and partition coefficient of L1; In K-L1>2.5). The findings reported here demonstrate the potential of ATPS to recover recombinant protein released from E coli by direct chemical extraction. (C) 2002 Society of Chemical Industry.
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