Improved production and purification of interferon-alpha-4a using a T7 RNA polymerase expression system.
An improved method for the synthesis and purification of human interferon-alpha-4a is presented. Interferon-alpha-4a was prepared using a T7 RNA polymerase expression system, where it was expressed from the vector pET-3a. Biologically active interferon-alpha-4a was isolated from the E. coli cells and purified using protamine sulphate precipitation, anion-exchange chromatography and affinity chromatography utilising a monoclonal antibody specific for human interferons-alpha.
Waine
G J
GJ
Department of Biochemistry, Monash University, Clayton, Victoria, Australia.
Wines
B D
BD
Wang
L
L
McMullen
G L
GL
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D015153:Blotting, Western; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D007370:Interferon Type I / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification*; D016898:Interferon-alpha; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011479:Protamines / Q000737:chemistry; D011994:Recombinant Proteins; D012293:Rifampin / Q000494:pharmacology; D014162:Transfection; D014764:Viral Proteins
D000911:Antibodies, Monoclonal; D007370:Interferon Type I; D016898:Interferon-alpha; D011479:Protamines; D011994:Recombinant Proteins; D014764:Viral Proteins; C068567:interferon alpha 4A; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D012293:Rifampin