Rapid high level production and purification of recombinant murine and human interferons alpha from Escherichia coli.
The availability of large quantities of pure interferon alpha (IFN-alpha) subtypes for in vivo studies has often proved difficult. This paper presents details on the use of the commercially available pGEX expression system for the production and purification of milligram (mg) quantities of recombinant Murine (Mu) and Human (Hu) IFNs-alpha-1 in Escherichia coli. Initially a fusion product is made which can be rapidly purified on a glutathione-sepharose 4B affinity matrix. Biologically active IFN-alpha can then be released from the matrix by cleavage with the restriction protease activated factor X (FXa+7,++). Routine yields of the final products were in the range of 0.5 to 2.0 mg/l of original culture.
Boyer
S J
SJ
Department of Microbiology, University of Western Australia, Queen Elizabeth II Medical Centre, Nedlands.
Colley
S M
SM
Lai
C M
CM
Swaminathan
N
N
Smith
W A
WA
Beilharz
M W
MW
D000818:Animals; D004926:Escherichia coli / Q000378:metabolism*; D006801:Humans; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000302:isolation & purification; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification
D016898:Interferon-alpha; D011993:Recombinant Fusion Proteins