Cellulose binding fusion proteins for immobilization and purification of polypeptides
AssigneeThe University of British Columbia
InventorsDouglas G. Kilburn, Robert C. Miller, Richard A. J. Warren, Neil R. Gilkes
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Abstract
A fusion protein is prepared containing a polypeptide such as an enzyme and an amino acid sequence having a substrate binding region of a polysaccharidase such as cellulase that has essentially no polysaccharidase activity. By contacting the fusion protein with an affinity matrix containing a substrate such as cellulose for the cellulase substrate binding region, the substrate binding region binds to the affinity matrix to immobilize the polypeptide. The polypeptide can be purified by separating the fusion protein or polypeptide from the affinity matrix.
Claims7
In a method using an affinity matrix for immobilization of a polypeptide of interest, the improvement which comprises:
- preparing a fusion protein comprising said polypeptide of interest and an amino acid sequence comprising a substrate binding region of a cellulose with the provisio that said amino acid sequence is essentially lacking in cellulose activity;
- and
- contacting said fusion protein with an affinity matrix comprising a cellulose substrate for said cellulose whereby said substrate binding region binds to said affinity matrix.
In a method using an affinity matrix for purification of a polypeptide of interest, the improvement which comprises:
- preparing a fusion protein comprising said polypeptide of interest and an amino acid sequence comprising a substrate binding region of a cellulose, with the proviso that said amino acid sequence is essentially lacing in cellulose activity;
- contacting said fusion protein with an affinity matrix comprising a cellulose substrate for said cellulose whereby said substrate binding region binds to said affinity matrix;
- and
- separating the fusion protein or the polypeptide of interest from the affinity matrix.
Dependencies are inferred from the claim text.