Purification of proteins employing CTAP-III fusions
AssigneeSRI International
InventorsPaul H. Johnson, Ping Sze, Richard C. Winant, Jerome B. Lazar
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Abstract
The present invention provides a process for the recovery of heterologous proteins from CTAP-III fusion proteins comprising expressing a fusion protein having a first amino acid sequence, a second amino acid sequence, and a selectable site which may be cleaved to provide first and second polypeptide fragments, respectively, wherein the first amino acid fragment is homologous to CTAP-III, and the first and second fragments have different pI values; cleaving the fusion protein to provide the first and second fragments; and separating the first and second fragments by ion exchange chromatography.
Claims4
In a method for purifying an heterologous polypeptide from a fusion polypeptide produced in a recombinant bacterial microorganism which method comprises the steps of expressing a fusion polypeptide composed of a connective tissue activating-peptide III polypeptide and the heterologous polypeptide separated by an intervening selectable cleavage site, separating the fusion polypeptide by cleaving at the selectable cleavage site, and subsequently recovering the heterologous polypeptide, said improvement employing a connective tissue activating-peptide III polypeptide having a selectable internal cleavage site whereby said separation step cleaves the fusion polypeptide and internally cleaves the connective tissue activating-peptide III polypeptide to produce connective tissue activating-peptide III peptides, each peptide having an isoelectric point difference of a minimum of approximately 1.5 pH units between the heterologous polypeptide and the connective tissue activating-peptide III peptide.
Dependencies are inferred from the claim text.