AI Article Synopsis
- A synthetic gene for human platelet factor 4 (hPF4) was successfully expressed in E. coli and processed into its active form (rhPF4) through cyanogen bromide treatment and column chromatography.
- The recombinant hPF4 maintains the same tetrameric structure, heparin-binding ability, and angiogenesis-inhibiting properties as the natural hPF4.
- Efforts to purify rhPF4 from residual fusion protein were challenging, leading to the development of a modified fusion strategy that can enhance purification methods for other recombinant proteins without affecting their expression levels.
Article 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.
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| http://dx.doi.org/10.1016/1046-5928(91)90062-n | DOI Listing |
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