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        The Use of Recombinant Fusion Proteases in the Affinity Purification of Recombinant Proteins

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        788
        At present the protein expression systems used commonly by researchers incorporate an affinity tail fused to the protein of interest. These affinity tails provide a convenient and efficient method for the purification of the expressed fusion protein using affinity chromatography. Many different affinity tails have been developed and a few of the commonly used fusion protein expression systems are listed in Table 1 . The plasmid expression vectors for the production of fusion proteins in various hosts are available commercially and with the advent of the polymerase chain reaction (PCR) any gene of known sequence can be cloned into any expression vector. Most affinity tails are linked to the N-terminus of the protein of interest, but some affinity tails are able to be linked to either the N- or C-terminus of the protein of interest. The choice of affinity tail to use for the expression of any particular protein is empirical since the factors leading to the high expression of recombinant proteins in foreign hosts have yet to be elucidated.
        Table 1  Systems Commonly Used for the Expression of Recombinant Fusion Proteins in E. coli a

        Affinity tail

        Elution ligand

        Supplier

        References

        Glutathione S-transferase

        Glutathione

        Pharmacra

        (5)

        Transitional metals

        Transrtional metals

        Qiagen, Inc

        (6,7)

        Bmdmg polypepttdes

        e.g,Zn2+ ,Cu2+ ,N1 2+

           

        Maltose bmdmg protein

        Maltose

        New England

        (8)

           

        Biolabs

         

        Staphylococcus aureus

        IgG

        Pharmacia

        (9)

        protein A

             

        Biotmylated pepttdes

        Streptavidin or

        Promega

        Promega

         

        avidin

           
        a Transrtronal metal brndlng polypepttdes can be fused to either the N or C terminus of proteins
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