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US4703008A Expired / Lapsed US ⏱️ 0.0 years remaining

DNA sequences encoding erythropoietin

Assignee: Amgen Inc
•
Filing: 1984-11-30
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Granted: 1987-10-27
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Expires: 2004-11-30
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Abstract

Abstract Disclosed are novel polypeptides possessing part or all of the primary structural conformation and one or more of the biological properties of mammalian erythropoietin ("EPO") which are characterized in preferred forms by being the product of procaryotic or eucaryotic host expression of an exogenous DNA sequence. Illustratively, genomic DNA, cDNA and manufactured DNA sequences coding for part or all of the sequence of amino acid residues of EPO or for analogs thereof are incorporated into autonomously replicating plasmid or viral vectors employed to transform or transfect suitable procaryotic or eucaryotic host cells such as bacteria, yeast or vertebrate cells in culture. Upon isolation from culture media or cellular lysates or fragments, products of expression of the DNA sequences display, e.g., the immunological properties and in vitro and in vivo biological activities of EPO of human or monkey species origins. Disclosed also are chemically synthesized polypeptides sharing the biochemical and immunological properties of EPO. Also disclosed are improved methods for the detection of specific single stranded polynucleotides in a heterologus cellular or viral sample prepared from, e.g., DNA present in a plasmid or viral-borne cDNA or genomic DNA "library".

Inventors

  • 👤 Fu-Kuen Lin

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-24 04:11
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/505; A; A61; A61P; A61P7/00; C07K16/00; C07K16/18; C07K16/22; C12; C12N; C12N15/00; C12N15/09; C12N15/63; C12N15/70; C12N15/79; C12N15/80; C12N15/81; C12N15/85; A61K; A61K38/00

Claims Analysis

Total Claims: 36
Claim 1
Independent Claim 48 words
A purified and isolated DNA sequence encoding erythropoietin, said DNA sequence selected from the group consisting of: (a) the DNA sequences set out in FIGS. 5 and 6 or their complementary strands; and (b) DNA sequences which hybridize under stringent conditions to the DNA sequences defined in (a).
Claim 2
Dependent Claim 48 words
A purified and isolated DNA sequence encoding erythropoietin, said DNA sequence selected from the group consisting of: (a) the DNA sequences set out in FIGS. 5 and 6 or their complementary strands; and (b) DNA sequences which hybridize under stringent conditions to the DNA sequences defined in (a).
Claim 3
Dependent Claim 15 words
A purified and isolated DNA sequence consisting essentially of a DNA sequence encoding human erythropoietin.
Claim 4
Dependent Claim 15 words
A purified and isolated DNA sequence consisting essentially of a DNA sequence encoding human erythropoietin.
Claim 5
Dependent Claim 15 words
A purified and isolated DNA sequence consisting essentially of a DNA sequence encoding monkey erythropoietin.
Claim 6
Dependent Claim 15 words
A purified and isolated DNA sequence consisting essentially of a DNA sequence encoding monkey erythropoietin.
Claim 7
Dependent Claim 30 words
A procaryotic or eucaryotic host cell transformed or transfected with a DNA sequence according to claim 1, 2 or 3 in a manner allowing the host cell to express erythropoietin.
Claim 8
Dependent Claim 20 words
A biologically functional circular plasmid or viral DNA vector including a DNA sequence according to claim 1, 2, or 3.
Claim 9
Dependent Claim 18 words
A procaryotic or eucaryotic host cell stably transformed or transfected with a DNA vector according to claim 5.
Claim 10
Dependent Claim 55 words
A purified and isolated DNA sequence consisting essentially of a DNA sequence encoding a polypeptide having an amino acid sequence sufficiently duplicative of that of erythropoietin to allow possession of the biological property of causing bone marrow cells to increase production of reticulocytes and red blood cells, and to increase hemoglobin synthesis or iron uptake.
Claim 11
Dependent Claim 55 words
A purified and isolated DNA sequence consisting essentially of a DNA sequence encoding a polypeptide having an amino acid sequence sufficiently duplicative of that of erythropoietin to allow possession of the biological property of causing bone marrow cells to increase production of reticulocytes and red blood cells, and to increase hemoglobin synthesis or iron uptake.
Claim 12
Dependent Claim 7 words
A cDNA sequence according to claim 7.
Claim 13
Dependent Claim 11 words
A monkey species erythropoietin coding DNA sequence according to claim 8.
Claim 14
Dependent Claim 18 words
A DNA sequence according to claim 9 and including the protein coding region set forth in FIG. 5.
Claim 15
Dependent Claim 8 words
A genomic DNA sequence according to claim 7.
Claim 16
Dependent Claim 11 words
A human species erythropoietin coding DNA sequence according to claim 11.
Claim 17
Dependent Claim 18 words
A DNA sequence according to claim 12 and including the protein coding region set forth in FIG. 6.
Claim 18
Dependent Claim 19 words
A DNA sequence according to claim 7 and including one or more codons preferred for expression in E.coli cells.
Claim 19
Dependent Claim 14 words
A DNA sequence according to claim 14, coding for expression of human species erythropoietin.
Claim 20
Dependent Claim 17 words
A DNA sequence according to claim 15 including the protein coding region set forth in FIG. 7.
Claim 21
Dependent Claim 19 words
A DNA sequence according to claim 7 and including one or more codons preferred for expression in yeast cells.
Claim 22
Dependent Claim 14 words
A DNA sequence according to claim 17, coding for expression of human species erythropoietin.
Claim 23
Dependent Claim 17 words
A DNA sequence according to claim 18 including the protein coding region set forth in FIG. 8.
Claim 24
Dependent Claim 14 words
A DNA sequence according to claim 7 covalently associated with a detectable label substance.
Claim 25
Dependent Claim 14 words
A DNA sequence according to claim 20 wherein the detectable label is a radiolabel.
Claim 26
Dependent Claim 8 words
A single-strand DNA sequence according to claim 20.
Claim 27
Dependent Claim 31 words
A procaryotic or eucaryotic host cell transformed or transfected with a DNA sequence according to claim 7, 8, or 11 in a manner allowing the host cell to express said polypeptide.
Claim 28
Dependent Claim 19 words
A transformed or transfected host cell according to claim 23 which host cell is capable of glycosylating said polypeptide.
Claim 29
Dependent Claim 11 words
A transformed or transfected mammalian host cell according to claim 24.
Claim 30
Dependent Claim 10 words
A transformed or transfected COS cell according to claim 25.
Claim 31
Dependent Claim 10 words
A transformed or transfected CHO cell according to claim 25.
Claim 32
Dependent Claim 17 words
A biologically functional circular plasmid or viral DNA vector including a DNA sequence according to claim 7.
Claim 33
Dependent Claim 18 words
A procaryotic or eucaryotic host cell stably transformed or transfected with a DNA vector according to claim 28.
Claim 34
Dependent Claim 37 words
A DNA sequence according to claim 7 coding for [Phe 15 ]hEPO, [Phe 49 ]hEPO, [Phe 145 ]hEPO, [His 7 ]hEPO, [Asn 2 des-Pro 2 through Ile 6 ]hEPO, [des-Thr 163 through Arg 166 hEPO, or [Δ27-55]hEPO.
Claim 35
Dependent Claim 22 words
A purified and isolated DNA sequence as set out in FIGS. 5 or 6 or the complementary strand of such a sequence.
Claim 36
Dependent Claim 22 words
A purified and isolated DNA sequence as set out in FIGS. 5 or 6 or the complementary strand of such a sequence.

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1990-10-26 1991-04-27 1991-10-27 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1994-10-26 1995-04-27 1995-10-27 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1998-10-26 1999-04-27 1999-10-27 $7700.00 Window Closed / Lapsed

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