DNA encoding erythropoietin multimers having modified 5′ and 3′ sequences and its use to prepare EPO therapeutics
AssigneeBeth Israel Deaconess Medical Center, Inc.
InventorArthur J. Sytkowski
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Abstract
Provided are nucleic acids encoding multimeric erythropoietin (EPO) proteins and having modifications in the 5′ and 3′ noncoding sequences relative to the corresponding sequences in native EPO DNA. The invention also relates to the use of such nucleic acids to produce multimeric EPO proteins, which may have altered activity as compared to EPO multimers expressed from nucleic acids having native 5′ and 3′ sequences.
Claims36
A nucleic acid molecule encoding a fusion protein, wherein the fusion protein comprises the amino acid sequence of two or more erythropoietin polypeptides, and wherein the nucleic acid molecule comprise a 5′ noncoding sequence comprising SEQ ID NOS:
- 3 or 4.
A nucleic acid molecule encoding a fusion protein, wherein the fusion protein comprises the amino acid sequence of two or more erythropoietin polypeptides, and wherein the nucleic acid molecule comprises a 3′ noncoding sequence comprising SEQ ID NO:
- 12.
A recombinant host cell comprising a nucleic acid molecule encoding a fusion protein, wherein the fusion protein comprises the amino acid sequence of two or more erythropoietin polypeptides, and wherein the nucleic acid molecule comprises a 5′ noncoding sequence comprising SEQ ID NOS:
- 3 or 4.
A recombinant host cell comprising a nucleic acid molecule encoding a fusion protein, wherein the fusion protein comprises the amino acid sequence of two or more erythropoietin polypeptides, and wherein the nucleic acid molecule comprises a 3′ noncoding sequence comprising SEQ ID NO:
- 12.
A method for making an erythropoietin fusion protein, comprising the steps of:
- a) transfecting a recombinant host cell with a vector comprising a nucleic acid molecule encoding a fusion protein, wherein the fusion protein comprises the amino acid sequence of two or more erythropoietin polypeptides, and wherein the nucleic acid molecule comprises a 5′ noncoding sequence comprising SEQ ID NOS:
- 3 or 4;
- and
- b) culturing the recombinant host cell in a suitable medium to produce the erythropoietin fusion protein.
A method for making an erythropoietin fusion protein, comprising the steps of:
- a) transfecting a recombinant host cell with a vector comprising a nucleic acid molecule encoding a fusion protein, wherein the fusion protein comprises the amino acid sequence of two or more erythropoietin polypeptides, and wherein the nucleic acid molecule comprises a 3′ noncoding sequence comprising SEQ ID NO:
- 12;
- and
- b) culturing the recombinant host cell in a suitable medium to produce the erythropoietin fusion protein.
A method for making an erythropoietin fusion protein, comprising the steps of:
- a) transfecting a recombinant host cell with a vector comprising a nucleic acid molecule encoding a fusion protein, wherein the fusion protein comprises the amino acid sequence of two or more erythropoietin polypeptides, and wherein the nucleic acid molecule comprises a 5′ noncoding sequence comprising SEQ ID NOS:
- 3 or 4 and further comprises a 3′ noncoding sequence comprising SEQ ID NO:
- 12;
- and
- b) culturing the recombinant host cell in a suitable medium to produce the erythropoietin fusion protein.
Dependencies are inferred from the claim text.