ES2576681T3 - Anticuerpos IL-1 alfa - Google Patents
Anticuerpos IL-1 alfa Download PDFInfo
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- ES2576681T3 ES2576681T3 ES09758739.8T ES09758739T ES2576681T3 ES 2576681 T3 ES2576681 T3 ES 2576681T3 ES 09758739 T ES09758739 T ES 09758739T ES 2576681 T3 ES2576681 T3 ES 2576681T3
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- human
- amino acid
- acid sequence
- mabp1
- light chain
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- 102000004125 Interleukin-1alpha Human genes 0.000 title 1
- 108010082786 Interleukin-1alpha Proteins 0.000 title 1
- 125000003275 alpha amino acid group Chemical group 0.000 abstract description 4
- 101001002634 Homo sapiens Interleukin-1 alpha Proteins 0.000 abstract description 2
- 239000013256 coordination polymer Substances 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 3
- 108010082117 matrigel Proteins 0.000 description 3
- 230000005012 migration Effects 0.000 description 3
- 238000013508 migration Methods 0.000 description 3
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 210000002469 basement membrane Anatomy 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- 108020004705 Codon Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 102100020881 Interleukin-1 alpha Human genes 0.000 description 1
- 108091081024 Start codon Proteins 0.000 description 1
- -1 Thr amino acids Chemical class 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 238000001879 gelation Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
- C07K16/244—Interleukins [IL]
- C07K16/245—IL-1
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P9/00—Drugs for disorders of the cardiovascular system
- A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/545—IL-1
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/069—Vascular Endothelial cells
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0693—Tumour cells; Cancer cells
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/10—Cells modified by introduction of foreign genetic material
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
- G01N33/6869—Interleukin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55522—Cytokines; Lymphokines; Interferons
- A61K2039/55527—Interleukins
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
- A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
- A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/52—Cytokines; Lymphokines; Interferons
- C07K14/54—Interleukins [IL]
- C07K14/5412—IL-6
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
- C07K2317/14—Specific host cells or culture conditions, e.g. components, pH or temperature
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
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- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
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- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
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- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/52—Assays involving cytokines
- G01N2333/54—Interleukins [IL]
- G01N2333/545—IL-1
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Abstract
Un AcM IgG1 humano purificado que se une específicamente a IL-1α humana, el AcM que comprende una cadena pesada unida covalentemente a una cadena ligera, en el que la cadena pesada comprende la secuencia de aminoácidos de SEQ ID NO: 9 y la cadena ligera comprende la secuencia de aminoácidos de SEQ ID NO: 11.
Description
>hIL-1a-IgG1-CP
>hIL-1a-IgG1-CP
La RV-CL se amplificó por PCR introduciendo los sitios HindIII/ClaI corriente arriba del codón de inicio ATG y un sitio
10 BsiWI en el extremo 3'. La secuencia de Kappa-CL constante humana se amplificó por PCR introduciendo un sitio BsiWI en el extremo 5' codificante de un Arg adicional, los primeros aminoácidos Thr, y un sitio BamHI en el extremo 3'. La secuencia de aminoácidos Kappa-CL constante humana correspondió a la entrada Swiss Prot P01834. Las secuencias RV-CP y Kappa-CL constante se ligaron utilizando el sitio BsiWI y se clonaron en pcDNA3 utilizando los sitios HindIII y BamHI.
15
7
>NATHMAB-hIL-1A-IGG1-CP
5 La secuencia completa codificante de NATHMAB-hIL-la/cadena ligera Kappa se sintetizó génicamente. La secuencia RV-CL correspondía a la secuencia de aminoácidos descrita en la patente de Estados Unidos 5.959.085. La secuencia de Kappa-CL constante humana correspondía a la entrada Swiss-Prot P01834. La secuencia de nucleótidos era un codón optimizado para la expresión en células CHO. Se añadió una secuencia de Kozac (gccacc) corriente arriba del ATG.
10 >NATHMAB-hIL-1A-K-CL
9
Cadena ligera
5 Ejemplo 6 -Afinidad de Unión a MABp1
La afinidad de unión de MABp1 purificado se determinó empleando resonancia superficial de plasmón (RSP) en un instrumento BIAcore 2000 (GE Health Sciences). Un Ac (Fc) IgG anti-humano monoclonal de ratón se inmovilizó covalentemente en las células de flujo de un chip sensor CM5 utilizando un kit de captura de Ac humano y un kit de 10 acoplamiento de amina (GE Health Sciences). Se alcanzaron normalmente niveles de inmovilización de 8.000
14.000 RU. Tras la inmovilización del Ac (Fc) de captura IgG anti-humano de ratón, se ejecutaron tres ciclos iniciales con tampón de migración HBS-EP (GE Health Sciences) y dos ciclos iniciales con MABp1 para estabilizar la superficie CM5 y eliminar cualquier Ac no unido covalentemente. Para el análisis, Ac MABp1 se diluyó en tampón de migración HBS-EP a una concentración final de 1 µg/ml y se inmovilizó a 700 RU en una célula de flujo del chip 15 sensor CM5. La citoquina IL-1A humana libre de transportador (eBioscience, n.º 34-8019) se diluyó en serie en tampón de migración HBS-EP en un intervalo de ensayo de 100 nM a 0,05 nM. El caudal era de 30 µl/min. Los datos de disociación para cada dilución de citoquinas se registraron durante 15 minutos. La superficie CM5 se regeneró después de cada ciclo empleando una inyección única de MgCl2 3 M durante 25 segundos a un caudal de 30 µl/min. El software BiaEvaluation y un modelo de unión de Langmuir se utilizaron para ajustar los datos. La KD para MABp1
20 determinó ser inferior a 2,0 x 10-10 M.
Ejemplo 7 -MABp1 inhibe la invasión de células tumorales de una matriz de la membrana basal
Matrigel (BD), una matriz de la membrana basal, se descongeló a 4 ºC durante la noche y el diluido (5 mg/ml a 1
25 mg/ml) en medio de cultivo celular frío libre de suero. 100 ul de matrigel diluido se colocaron en las cámaras superiores de un cultivo polarizado de 24 pocillos (Costar) y el cultivo polarizado se incubó a 37 ºC durante al menos 4 a 5 h para gelificación. Las células tumorales (MDA-MB-231 y THP-1) se recogieron de los matraces de cultivo tisular con tripsina/EDTA, se lavaron con medios de cultivo, y se volvieron a suspender en un medio que contenía SFB al 1 % en una densidad de 1 X 106 células/ml. El matrigel gelificado se lavó gradualmente con los medios de
30 cultivo libres de suero calentado, y se añadieron 100 ul de la suspensión celular a cada pocillo. La cámara inferior del cultivo polarizado se llenó con 600 ul de medio de cultivo, y las placas se incubaron a 37 ºC durante 12 a 24 h.
12
Claims (1)
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imagen1
Applications Claiming Priority (7)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US121391P | 1999-02-25 | ||
US5758608P | 2008-05-30 | 2008-05-30 | |
US57586P | 2008-05-30 | ||
US12139108P | 2008-12-10 | 2008-12-10 | |
US17835009P | 2009-05-14 | 2009-05-14 | |
US178350P | 2009-05-14 | ||
PCT/US2009/003355 WO2009148575A1 (en) | 2008-05-30 | 2009-06-01 | Interleukin-1 alpha abs and methods of use |
Publications (1)
Publication Number | Publication Date |
---|---|
ES2576681T3 true ES2576681T3 (es) | 2016-07-08 |
Family
ID=41380314
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
ES09758739.8T Active ES2576681T3 (es) | 2008-05-30 | 2009-06-01 | Anticuerpos IL-1 alfa |
ES16164746T Active ES2797126T3 (es) | 2008-05-30 | 2009-06-01 | Usos de anticuerpos IL-1 alfa |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
ES16164746T Active ES2797126T3 (es) | 2008-05-30 | 2009-06-01 | Usos de anticuerpos IL-1 alfa |
Country Status (24)
Country | Link |
---|---|
US (14) | US8034337B2 (es) |
EP (3) | EP3085773B1 (es) |
JP (5) | JP5670318B2 (es) |
KR (3) | KR20150127300A (es) |
CN (4) | CN103656638B (es) |
AU (1) | AU2009255670B2 (es) |
BR (1) | BRPI0909610B1 (es) |
CA (2) | CA3011485A1 (es) |
CO (1) | CO6351749A2 (es) |
DK (1) | DK2285409T3 (es) |
ES (2) | ES2576681T3 (es) |
HK (2) | HK1159481A1 (es) |
HU (1) | HUE029003T2 (es) |
IL (5) | IL209576A (es) |
MX (1) | MX2010012963A (es) |
MY (1) | MY154067A (es) |
NZ (1) | NZ590033A (es) |
PH (1) | PH12014501773A1 (es) |
PL (2) | PL2285409T3 (es) |
PT (1) | PT2285409T (es) |
RU (2) | RU2498998C2 (es) |
SI (1) | SI2285409T1 (es) |
WO (1) | WO2009148575A1 (es) |
ZA (1) | ZA201009224B (es) |
Families Citing this family (440)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20070084897A1 (en) | 2003-05-20 | 2007-04-19 | Shelton Frederick E Iv | Articulating surgical stapling instrument incorporating a two-piece e-beam firing mechanism |
US9060770B2 (en) | 2003-05-20 | 2015-06-23 | Ethicon Endo-Surgery, Inc. | Robotically-driven surgical instrument with E-beam driver |
US11896225B2 (en) | 2004-07-28 | 2024-02-13 | Cilag Gmbh International | Staple cartridge comprising a pan |
US8215531B2 (en) | 2004-07-28 | 2012-07-10 | Ethicon Endo-Surgery, Inc. | Surgical stapling instrument having a medical substance dispenser |
JP5645361B2 (ja) * | 2005-08-02 | 2014-12-24 | エクスバイオテク インコーポレーティッド | IL−1α自己抗体を用いた血管障害の診断、処置、および予防 |
US9237891B2 (en) | 2005-08-31 | 2016-01-19 | Ethicon Endo-Surgery, Inc. | Robotically-controlled surgical stapling devices that produce formed staples having different lengths |
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- 2013-02-08 US US13/762,453 patent/US8679489B2/en active Active
- 2013-02-14 US US13/767,292 patent/US8784817B2/en active Active
- 2013-02-17 IL IL224752A patent/IL224752A/en active IP Right Grant
- 2013-08-19 RU RU2013138469A patent/RU2666915C2/ru active
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2014
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- 2014-08-06 PH PH12014501773A patent/PH12014501773A1/en unknown
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- 2014-12-11 US US14/567,072 patent/US9181338B2/en active Active
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2015
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2017
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2019
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2020
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2021
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2023
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