Investigation on the purification and expression of cell I — Hep I recombinant FN polypeptide inE. Coli

Abstract

An anti-metastatic polypeptide, bifunctional-domain (Cell I -Hep I) recombinant polypeptide of human fibronectin, was expressedin E. coli and purified. The expression level was found to be about 20%– 30 % of the total cell proteins. In BL21 (DE3)/T7, anE. coli expressing system, lactose can be used as an inducer to substitute IPTG thereby reducing the cost by several hundredfold and it is suitable for the large-scale preparation of recombinant FN polypeptide. Cell I -Hep 1 fragment is an alkaline polypeptide. In BL21(DE3)/T7 expressing system, better isolation was achieved if DEAE-52, instead of CM-52, was used for ion-exchange chromatography. The purified product was obtained after heparin-agarose affinity chromatography following ion-exchange chromatography.

Keywords

fibronectin / recombinant polypeptide / induced expression / purification / metastasis

References

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