title: Expression and purification of active recombinant platelet factor 4 from a cleavable fusion protein.
abstract: A synthetic gene for human platelet factor 4 (hPF4) has been expressed at high levels as a fusion protein in Escherichia coli. The hPF4 sequence has been cleaved from the fusion protein by cyanogen bromide treatment and purified by column chromatography. Like hPF4, our recombinant hPF4 (rhPF4) is tetrameric under physiological conditions, binds heparin, and inhibits angiogenesis. Extensive purification to remove trace amounts of uncleaved fusion protein completely from the desired product rhPF4 was difficult. We have exploited recombinant DNA technology by modifying the fusion moiety to accomplish separation. This type of modification, which did not affect expression level, could be applied to other recombinant fusion proteins.
authors.affiliation: Repligen Corporation, Cambridge, Massachusetts 02139.
authors.forename: J A
authors.initials: JA
authors.lastname: Myers
authors.forename: G S
authors.initials: GS
authors.lastname: Gray
authors.forename: D J
authors.initials: DJ
authors.lastname: Peters
authors.forename: R J
authors.initials: RJ
authors.lastname: Grimaila
authors.forename: A J
authors.initials: AJ
authors.lastname: Hunt
authors.forename: T E
authors.initials: TE
authors.lastname: Maione
authors.forename: W T
authors.initials: WT
authors.lastname: Mueller
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002847:Chromatography, Agarose; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D010978:Platelet Factor 4 / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
chemical_list: D011993:Recombinant Fusion Proteins; D010978:Platelet Factor 4; D004247:DNA