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Fractionation of Recombinant Tumor Necrosis Factor Using Hydrophobic and Hydrophilic Membranes
Author(s) -
Dorin Glenn,
Thomson James,
Hanisch Wolf
Publication year - 1990
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp00006a013
Subject(s) - membrane , recombinant dna , chemistry , fractionation , yield (engineering) , filtration (mathematics) , tumor necrosis factor alpha , size exclusion chromatography , intracellular , escherichia coli , chromatography , cell disruption , biochemistry , biology , materials science , enzyme , immunology , metallurgy , gene , statistics , mathematics
Abstract Recombinant tumor necrosis factor (TNF) is expressed in Escherichia coli as a soluble intracellular protein. A purification process is described that incorporates a hydrophilic membrane (cellulosic) separation followed by a hydrophobic one (PTFE). The hydrophilic step is a traditional one in that species are separated primarily on the basis of size. The hydrophobic step separates species on the basis of parameters apparently not related to size. By combining these two steps, an increase in TNF purity of 7‐10‐fold can be achieved with a yield of 50%. The effects of cellular debris and pH on selectivity and recovery of the hydrophobic filtration step are discussed.