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US 5,856,456 AGranted
Linker for linked fusion polypeptides
AssigneeEnzon, Inc.
InventorsMarc D. Whitlow, David R. Filpula
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AssigneeEnzon, Inc.
InventorsMarc D. Whitlow, David R. Filpula
Estimating the term…
The invention is directed to a novel peptide linker useful for connecting polypeptide constituents into a novel linked fusion polypeptide. The peptide linker of the invention provides greater stability and is less susceptible to aggregation than previously known peptide linkers. The peptide linker of the invention may be up to about 50 amino acids in length and contains at least one occurrence of a charged amino acid followed by a proline. When used for making a single chain Fv (sFv), the peptide linker is preferably from 18 to about 30 amino acids in length. A preferred embodiment of the peptide linker of the invention comprises the sequence: GSTSGSGXPGSGEGSTKG (SEQ. ID NO 1), where X is a charged amino acid, preferably lysine or arginine. Methods of making linked fusion polypeptides using the peptide linker of the invention are claimed. DNA molecules encoding such linked fusion polypeptides, and methods of producing such linked fusion polypeptides from these DNA molecules are also claimed.
A DNA molecule encoding a linked fusion polypeptide comprising a first polypeptide and a second polypeptide connected by a peptide linker, wherein said peptide linker is 18 to 50 amino acid residues long, said peptide linker comprises one or more occurrences of the sequence XP wherein X is a charged amino acid, and said peptide linker further comprises the sequence represented by GSTSGSGXPGSGSGEGSTKG (SEQ, ID No. 1), said sequence positioned within said peptide linker so as to inhibit proteolysis of said linker by either subtilisin or typsin.
A DNA molecule encoding the polypeptide comprising the sequence represented by GSTSGSGXPGSGEGSTKG (SEQ. ID No.1) wherein X is a charged amino acid.
A DNA molecule encoding a linked fusion polypeptide comprising a first polypeptide and a second polypeptide connected by a peptide linker, wherein said first and second polypeptides are derived from the same multi-chain protein, said multi-chain protein being a member of the immunoglobulin superfamily;
A DNA molecule encoding a linked fusion polypeptide comprising a first polypeptide and a second polypeptide connected by a peptide linker, wherein said peptide linker is 18 to 50 amino acid residues long, said peptide linker comprises one or more occurrences of a sequence XP wherein is a charged amino acid, and said peptide linker further comprises the sequence represented by GSTSGSGKPGSGEGSTKG (SEQ. ID No. 10), wherein said sequence is positioned within said peptide linker so as to inhibit proteolysis of said linker by either subtilisin or trypsin.
A DNA molecule encoding a linked fusion polypeptide comprising a first polypeptide and a second polypeptide connected by a peptide linker, wherein said peptide linker is 12 to 50 amino acid residues long, said peptide linker comprises one or more occurrences of a sequence XP wherein X is a charged amino acid, and said peptide linker further comprises the sequence represented by GSTSGKPSEGKG (amino acid nos. 113-124 of SEQ. ID No. 12), wherein said sequence is positioned within said peptide linker so as to inhibit proteolysis of said linker by at least subtilisin or trypsin.
A DNA molecule encoding a linked fusion polypeptide comprising a first polypeptide and a second polypeptide connected by a peptide linker, wherein said peptide linker is 12 to 50 amino acid residues long, said peptide linker comprises one or more occurrences of the sequence XP wherein X is a charged amino acid, and said peptide linker further comprises the sequence represented by GSTSGXPSEGKG (SEQ. ID NO 25), wherein said sequence is positioned within said peptide linker so as to inhibit proteolysis of said linker by either subtilisin or trypsin.
A DNA molecule encoding the polypeptide comprising the sequence represented by GSTSGSGKPGSGEGSTKG (SEQ. ID No. 10).
A DNA molecule encoding the polypeptide comprising the sequence represented by GSTSGKPSEGKG (amino acid nos. 113-124 of SEQ. ID No. 12).
A DNA molecule encoding the polypeptide comprising the sequence represented by GSTSGXPSEGKG (SEQ. ID NO 25) wherein X is a charged amino acid.
Dependencies are inferred from the claim text.