WO2009118460A1 - Lapins transgeniques producteurs de facteur vii humain - Google Patents
Lapins transgeniques producteurs de facteur vii humain Download PDFInfo
- Publication number
- WO2009118460A1 WO2009118460A1 PCT/FR2008/000396 FR2008000396W WO2009118460A1 WO 2009118460 A1 WO2009118460 A1 WO 2009118460A1 FR 2008000396 W FR2008000396 W FR 2008000396W WO 2009118460 A1 WO2009118460 A1 WO 2009118460A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- factor vii
- human factor
- transgenic
- milk
- rabbits
- Prior art date
Links
- 241000283973 Oryctolagus cuniculus Species 0.000 title claims abstract description 35
- 230000009261 transgenic effect Effects 0.000 title claims abstract description 28
- 229940099816 human factor vii Drugs 0.000 title claims abstract description 23
- 235000013336 milk Nutrition 0.000 claims abstract description 20
- 210000004080 milk Anatomy 0.000 claims abstract description 20
- 239000008267 milk Substances 0.000 claims abstract description 19
- 238000004519 manufacturing process Methods 0.000 claims abstract description 10
- 239000002994 raw material Substances 0.000 claims abstract description 3
- 108700019146 Transgenes Proteins 0.000 claims description 17
- 241000124008 Mammalia Species 0.000 claims description 3
- 108091033319 polynucleotide Proteins 0.000 claims description 3
- 102000040430 polynucleotide Human genes 0.000 claims description 3
- 239000002157 polynucleotide Substances 0.000 claims description 3
- 230000022532 regulation of transcription, DNA-dependent Effects 0.000 claims description 2
- 210000004907 gland Anatomy 0.000 claims 1
- 210000005075 mammary gland Anatomy 0.000 abstract description 8
- 102100023804 Coagulation factor VII Human genes 0.000 description 17
- 108010023321 Factor VII Proteins 0.000 description 17
- 229940012413 factor vii Drugs 0.000 description 17
- 108090000623 proteins and genes Proteins 0.000 description 16
- 238000013518 transcription Methods 0.000 description 11
- 230000035897 transcription Effects 0.000 description 11
- 210000002257 embryonic structure Anatomy 0.000 description 10
- 239000003623 enhancer Substances 0.000 description 10
- 239000013598 vector Substances 0.000 description 10
- 235000018102 proteins Nutrition 0.000 description 9
- 102000004169 proteins and genes Human genes 0.000 description 9
- 101710087237 Whey acidic protein Proteins 0.000 description 8
- 241001465754 Metazoa Species 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 7
- 108010076119 Caseins Proteins 0.000 description 6
- 238000000520 microinjection Methods 0.000 description 6
- 102000011632 Caseins Human genes 0.000 description 5
- 108091092195 Intron Proteins 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 241000894007 species Species 0.000 description 5
- 108020004414 DNA Proteins 0.000 description 4
- 108091005904 Hemoglobin subunit beta Proteins 0.000 description 4
- 108090000144 Human Proteins Proteins 0.000 description 4
- 102000003839 Human Proteins Human genes 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 230000004481 post-translational protein modification Effects 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 108010054265 Factor VIIa Proteins 0.000 description 3
- 101001035782 Gallus gallus Hemoglobin subunit beta Proteins 0.000 description 3
- 230000004988 N-glycosylation Effects 0.000 description 3
- 230000004989 O-glycosylation Effects 0.000 description 3
- 108010076504 Protein Sorting Signals Proteins 0.000 description 3
- 101150059663 WAP gene Proteins 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 229940024606 amino acid Drugs 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000003776 cleavage reaction Methods 0.000 description 3
- 238000010367 cloning Methods 0.000 description 3
- 230000006251 gamma-carboxylation Effects 0.000 description 3
- 239000005556 hormone Substances 0.000 description 3
- 229940088597 hormone Drugs 0.000 description 3
- 230000010354 integration Effects 0.000 description 3
- 210000001161 mammalian embryo Anatomy 0.000 description 3
- 230000013011 mating Effects 0.000 description 3
- 210000003101 oviduct Anatomy 0.000 description 3
- 210000002381 plasma Anatomy 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 238000003752 polymerase chain reaction Methods 0.000 description 3
- 230000007017 scission Effects 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- YGEHCIVVZVBCLE-FSYGUOKUSA-N 3alpha-galactobiose Chemical group OC[C@@H](O)[C@H](O)[C@@H]([C@@H](O)C=O)O[C@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O YGEHCIVVZVBCLE-FSYGUOKUSA-N 0.000 description 2
- 102000004506 Blood Proteins Human genes 0.000 description 2
- 108010017384 Blood Proteins Proteins 0.000 description 2
- 102100022641 Coagulation factor IX Human genes 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 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 2
- 108010062466 Enzyme Precursors Proteins 0.000 description 2
- 102000010911 Enzyme Precursors Human genes 0.000 description 2
- 108010076282 Factor IX Proteins 0.000 description 2
- 208000009292 Hemophilia A Diseases 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 108010000521 Human Growth Hormone Proteins 0.000 description 2
- 102000014171 Milk Proteins Human genes 0.000 description 2
- 108010011756 Milk Proteins Proteins 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 2
- 238000002105 Southern blotting Methods 0.000 description 2
- 108010000499 Thromboplastin Proteins 0.000 description 2
- 102000002262 Thromboplastin Human genes 0.000 description 2
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 2
- 229960000723 ampicillin Drugs 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 2
- 235000021240 caseins Nutrition 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 239000000539 dimer Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229960004222 factor ix Drugs 0.000 description 2
- 229940012414 factor viia Drugs 0.000 description 2
- 230000002008 hemorrhagic effect Effects 0.000 description 2
- 238000005805 hydroxylation reaction Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 235000021239 milk protein Nutrition 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 230000001360 synchronised effect Effects 0.000 description 2
- 238000011830 transgenic mouse model Methods 0.000 description 2
- 238000013519 translation Methods 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- AUTALUGDOGWPQH-UBLOVXTBSA-N (2r,3s,4r,5r)-2,3,4,5,6-pentahydroxyhexanal;(2r,3s,4r)-2,3,4,5-tetrahydroxypentanal Chemical compound OC[C@@H](O)[C@H](O)[C@@H](O)C=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O AUTALUGDOGWPQH-UBLOVXTBSA-N 0.000 description 1
- ODDPRQJTYDIWJU-UHFFFAOYSA-N 3'-beta-D-galactopyranosyl-lactose Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(OC2C(OC(O)C(O)C2O)CO)OC(CO)C1O ODDPRQJTYDIWJU-UHFFFAOYSA-N 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 208000031872 Body Remains Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 108091029523 CpG island Proteins 0.000 description 1
- 108010054218 Factor VIII Proteins 0.000 description 1
- 102000001690 Factor VIII Human genes 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 108010049003 Fibrinogen Proteins 0.000 description 1
- 102000008946 Fibrinogen Human genes 0.000 description 1
- 102000030902 Galactosyltransferase Human genes 0.000 description 1
- 108060003306 Galactosyltransferase Proteins 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 102100021519 Hemoglobin subunit beta Human genes 0.000 description 1
- 208000031220 Hemophilia Diseases 0.000 description 1
- 101500025568 Homo sapiens Saposin-D Proteins 0.000 description 1
- 102000002265 Human Growth Hormone Human genes 0.000 description 1
- 239000000854 Human Growth Hormone Substances 0.000 description 1
- 241000598436 Human T-cell lymphotropic virus Species 0.000 description 1
- 206010020460 Human T-cell lymphotropic virus type I infection Diseases 0.000 description 1
- 241000714260 Human T-lymphotropic virus 1 Species 0.000 description 1
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical group C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 description 1
- 108010063045 Lactoferrin Proteins 0.000 description 1
- 102000010445 Lactoferrin Human genes 0.000 description 1
- 108091026898 Leader sequence (mRNA) Proteins 0.000 description 1
- 241000713333 Mouse mammary tumor virus Species 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 101800004937 Protein C Proteins 0.000 description 1
- 102000017975 Protein C Human genes 0.000 description 1
- 108010094028 Prothrombin Proteins 0.000 description 1
- 102100027378 Prothrombin Human genes 0.000 description 1
- 108020005067 RNA Splice Sites Proteins 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 101800001700 Saposin-D Proteins 0.000 description 1
- 208000006011 Stroke Diseases 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 206010042573 Superovulation Diseases 0.000 description 1
- 108090000190 Thrombin Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 229930003448 Vitamin K Natural products 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 101150087698 alpha gene Proteins 0.000 description 1
- 125000003169 alpha-Gal epitope group Chemical group [C@H]1([C@H](O)[C@@H](O)[C@@H](O)[C@H](O1)CO)O[C@@H]1[C@H]([C@@H](O[C@@H]([C@@H]1O)CO)O[C@H]1[C@@H]([C@H](C(O[C@@H]1CO)*)NC(C)=O)O)O 0.000 description 1
- 235000020244 animal milk Nutrition 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 125000000637 arginyl group Chemical group N[C@@H](CCCNC(N)=N)C(=O)* 0.000 description 1
- 235000009582 asparagine Nutrition 0.000 description 1
- 150000001508 asparagines Chemical class 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N aspartic acid group Chemical group N[C@@H](CC(=O)O)C(=O)O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 108010006025 bovine growth hormone Proteins 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 230000035602 clotting Effects 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 150000001945 cysteines Chemical group 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000000368 destabilizing effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000013020 embryo development Effects 0.000 description 1
- 108010030074 endodeoxyribonuclease MluI Proteins 0.000 description 1
- 210000003989 endothelium vascular Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 229960000301 factor viii Drugs 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 229940012952 fibrinogen Drugs 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 230000003325 follicular Effects 0.000 description 1
- 230000008217 follicular development Effects 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000002695 general anesthesia Methods 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 150000002307 glutamic acids Chemical group 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 229940100689 human protein c Drugs 0.000 description 1
- 230000033444 hydroxylation Effects 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 238000000126 in silico method Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 229940090044 injection Drugs 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 230000009027 insemination Effects 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 229960000310 isoleucine Drugs 0.000 description 1
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 1
- 125000000741 isoleucyl group Chemical group [H]N([H])C(C(C([H])([H])[H])C([H])([H])C([H])([H])[H])C(=O)O* 0.000 description 1
- CSSYQJWUGATIHM-IKGCZBKSSA-N l-phenylalanyl-l-lysyl-l-cysteinyl-l-arginyl-l-arginyl-l-tryptophyl-l-glutaminyl-l-tryptophyl-l-arginyl-l-methionyl-l-lysyl-l-lysyl-l-leucylglycyl-l-alanyl-l-prolyl-l-seryl-l-isoleucyl-l-threonyl-l-cysteinyl-l-valyl-l-arginyl-l-arginyl-l-alanyl-l-phenylal Chemical compound C([C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 CSSYQJWUGATIHM-IKGCZBKSSA-N 0.000 description 1
- 229940078795 lactoferrin Drugs 0.000 description 1
- 235000021242 lactoferrin Nutrition 0.000 description 1
- 238000002350 laparotomy Methods 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 210000000287 oocyte Anatomy 0.000 description 1
- XNOPRXBHLZRZKH-MQYCRUOZSA-N oxytocin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CSSC[C@H](N)C(=O)N1)C(=O)N1C(CCC1)C(=O)N[C@@H](CC(C)C)C(=O)NCC(N)=O)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 XNOPRXBHLZRZKH-MQYCRUOZSA-N 0.000 description 1
- 229940082408 oxytocin injection Drugs 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 230000032696 parturition Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 description 1
- 239000013600 plasmid vector Substances 0.000 description 1
- 239000002797 plasminogen activator inhibitor Substances 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 229960000856 protein c Drugs 0.000 description 1
- 230000006337 proteolytic cleavage Effects 0.000 description 1
- 229940039716 prothrombin Drugs 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 235000004400 serine Nutrition 0.000 description 1
- 150000003355 serines Chemical group 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 229960004072 thrombin Drugs 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000012250 transgenic expression Methods 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 235000019168 vitamin K Nutrition 0.000 description 1
- 239000011712 vitamin K Substances 0.000 description 1
- 150000003721 vitamin K derivatives Chemical class 0.000 description 1
- 229940046010 vitamin k Drugs 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
- A01K67/0275—Genetically modified vertebrates, e.g. transgenic
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/04—Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
-
- 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
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- 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
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6424—Serine endopeptidases (3.4.21)
- C12N9/6437—Coagulation factor VIIa (3.4.21.21)
-
- 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
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
- C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
- C12N9/6424—Serine endopeptidases (3.4.21)
- C12N9/647—Blood coagulation factors not provided for in a preceding group or according to more than one of the proceeding groups
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/21—Serine endopeptidases (3.4.21)
- C12Y304/21021—Coagulation factor VIIa (3.4.21.21)
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/05—Animals comprising random inserted nucleic acids (transgenic)
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/107—Rabbit
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/01—Animal expressing industrially exogenous proteins
-
- 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
- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/008—Vector systems having a special element relevant for transcription cell type or tissue specific enhancer/promoter combination
-
- 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
- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/30—Vector systems having a special element relevant for transcription being an enhancer not forming part of the promoter region
-
- 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
- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/40—Vector systems having a special element relevant for transcription being an insulator
-
- 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
- C12N2830/00—Vector systems having a special element relevant for transcription
- C12N2830/80—Vector systems having a special element relevant for transcription from vertebrates
- C12N2830/85—Vector systems having a special element relevant for transcription from vertebrates mammalian
Definitions
- the present invention relates to the production of recombinant human factor VII in the milk of transgenic rabbits.
- Factor VII is a plasma protein involved in the process of blood coagulation, and in particular in the initiation of the extrinsic coagulation pathway.
- Factor VII which is a vitamin K-dependent glycoprotein, is synthesized in the liver as a 466 amino acid precursor comprising a signal peptide (residues 1-20) and a propeptide (residues 21-60).
- Factor VII circulating in blood plasma is a zymogen (or proenzyme) consisting of a peptide chain of 53 kDa containing 406 residues.
- Factor VII is activated by factor VIIa by proteolytic cleavage between arginine at position 152 and isoleucine at position 153:
- Factor VIIa is composed of a 152 amino acid light chain derived from the N-terminus of a molecular weight of about 20 kDa and a 254 amino acid heavy chain derived from the C-terminus of a molecular weight of about 30 kDa, which are covalently linked together by a disulfide bridge between the cysteines in position 135 and 262.
- Circulating factor VII may complex with the tissue factor (TF) produced by subendothelial fibroblasts, when this is released during a breach in the vascular endothelium.
- TF tissue factor
- the formation of this complex is accompanied by the activation of factor VII factor VIIa.
- the TF-VIIa complex activates factors IX and X, resulting in the formation of factors IXa and Xa, which in turn activate the conversion of prothrombin to thrombin, which allows the conversion of fibrinogen to fibrin, resulting in clot formation.
- the factor VlI / VIIa has been used for many years to treat patients with different haemostasis disorders (for example hemophilia type A, which corresponds to a deficiency of factor VIII, haemophilia type B which corresponds to a deficiency in factor IX, or hereditary deficits in factor VlI / VIIa), as well as hemorrhagic accidents of various origins such as cerebrovascular accidents, or hemorrhagic traumas.
- haemostasis disorders for example hemophilia type A, which corresponds to a deficiency of factor VIII, haemophilia type B which corresponds to a deficiency in factor IX, or hereditary deficits in factor VlI / VIIa
- hemorrhagic accidents of various origins such as cerebrovascular accidents, or hemorrhagic traumas.
- the factor VII preparations used were obtained from human plasmas.
- recombinant factor VII preparations produced by transformed mammalian cells are presently preferred. for expressing human factor VII, as described, for example, in Application EP 0 200 421.
- An alternative to the production of proteins of therapeutic interest by transformed cell cultures is their production by transgenic animals, and in particular in milk of these. This approach theoretically has many advantages, including a higher yield, a lower production cost than cell culture production, and an increase in production more easily and more flexible at the industrial level.
- these factors include the ability of the host species to produce in industrializable quantities, proteins with post-translational modifications similar to those of the native human protein. This factor is particularly critical if it is envisaged to produce in recombinant form in milk proteins such as factor VII, whose post-translational modifications are multiple and varied (cleavage of the signal peptide and the propeptide, ⁇ -carboxylation, ⁇ - hydroxylation, O-glycosylation and N-glycosylation), which are naturally produced in the liver. Indeed, the cells of the mammary gland have different post-translational modification capacities than those of the liver cells.
- transgenic mice expressing human protein C in their mammary glands could not properly perform propeptide cleavage and gamma-carboxylation (Drohan et al., Transgenic Res., 355-64, 1994). . Similar observations have been made in transgenic pigs for both protein C and factor IX (Lee et al., J. Biochem., 118, 81-7, 1995, Van Cott et al, Genet Anal., 15, 155). -60, 1999).
- the O-glycosylation and the N-glycosylation of the proteins can vary, qualitatively and quantitatively depending on the mammalian species concerned.
- the human proteins do not comprise a galactose-galactose ⁇ (1-3) galactose unit (Gal ⁇ 1-3Gal motif), since the gene coding for alpha-1,3-galactosyltransferase, responsible for the synthesis of this motif, is inactivated in humans and Old World monkeys (Europe, Asia) while present in other mammals.
- This motif (also called ⁇ -Gal epitope) is consequently highly immunogenic in humans (GALILI et al, Blood, 82 (8): 2485-2493, 1993).
- a glycosylated human protein produced in recombinant form in mammals possessing an active alpha 1,3 galactosyltransferase is therefore likely to carry a large amount of Gal ⁇ 1-3Gal motifs and may cause significant undesirable immune reactions.
- transgenic rabbits expressing in their mammary glands a sequence coding for human factor VII produced in their milk a correctly cleaved recombinant factor VII, and furthermore not containing or containing only very few Gal ⁇ l-3Gal patterns.
- the present invention therefore relates to a transgenic rabbit expressing a human factor VII in its milk.
- a transgenic rabbit according to the invention contains in its genome one or more copies of a transgene comprising a polynucleotide encoding a human factor VII, placed under the transcriptional control of a promoter allowing its specific expression in the mammary gland cells of said rabbit. .
- transgene is meant a nucleic acid construct stably inserted into the genome of a host organism, which is transmitted to its offspring from generation to generation.
- the transgene allows expression of a protein of interest (factor VII) in the transgenic host animal milk.
- Promoters for the specific expression in mammary gland cells of the rabbit are known per se.
- they may be promoters of casein genes or milk serum proteins: mention will be made in particular of the ⁇ , ⁇ , or K casein promoters, the ⁇ -lactoglobin promoter, the P-lactalbumin promoter and the WAP (whey acidic protein) promoter; , or that of lactoferrin.
- It may be a promoter from the rabbit, such as the WAP promoter described in Application EP0527063, or a promoter from another mammalian species.
- the polynucleotide encoding human factor VII is preferably a cDNA, or the coding portion (ORF) thereof. It can be the natural cDNA of factor VII, the sequence of which is known per se, and available on the databases, for example under accession number Ml 3232, or a sequence coding for a modified human factor VII variant in particular to increase its activity and / or to eliminate its undesirable effects; by way of examples, mention may be made of the variants described in Application 2008/0010693 or that described in the publication by Sorensen et al. (Br. J. HaematoL, 137 (2): 158-65, 2007). It is advantageous to use a DNA sequence optimized for expression in the mammary glands of rabbits.
- Such a sequence can be obtained in silico by techniques well known to those skilled in the art, in order to eliminate cryptic splice sites, A / T rich sequences, destabilizing mRNAs, polyadenylation sites as well as that potentially parasitic TATA boxes, and CpG islands, and optimize the codons to reflect the preferences of the mammary gland cells of rabbits to produce milk proteins.
- the transgene contains, besides the promoter and the coding sequence for factor VII, other elements intended to optimize the transcription and / or the translation of the recombinant protein.
- elements are known in themselves to those skilled in the art (see, for example, Houdebine et ah, in: Cari A. Pinkert (ed), Vector Design for Transgene Transgenic Expression Animal Technology, 2nd edn, New York Academy Press, 419-458, 2002).
- a strong isolator placed at 5 'of the promoter, guaranteeing a level of expression of the protein dependent on the number of transgene copies integrated and independent of the site of integration of the transgene into the genome of the animal: for example the 5 ⁇ S4 region of the chicken beta-globin gene (Taboit-Dameron et al., Transgenic Res., 8: 223-235, 1999; Rival-Gervier et al., Transgenic Res., 12: 723). - 730, 2003).
- exon / intron pairs which may contain one or more transcription or translation enhancers: by way of examples, mention may be made of: the introns of the late and early genomes of the SV40 virus genome, the first intron of a beta-globin gene, EF1 alpha gene introns, Palpha-sl casein introns, WAP gene introns, human and bovine growth hormone genes introns; the "enhancer” sequences can in particular be chosen from those present in the LTR sequences of the HTLV virus, or the MMTV virus (murine mammary tumor virus), the "enhancer” sequence of the immunoglobin gene, the “enhancer” sequence of the gene alpha-1 casein, the "enhancer” sequence of beta-globin.
- the "enhancer” sequence may also be the distal region upstream (up to 140 kbp) of the WAP gene or the downstream distal region (at least 10 kbp) of the WAP gene, as described by Rival-Gervier and al. (Mol Reprod Develop, 63: 161-167, 2002), in Application EP 1 217 071
- terminators of the SV40 early or late genes those of the beta-globin genes, WAP genes, human or bovine growth.
- Transgenesis can be carried out by conventional methods known per se.
- the transgene is introduced by micro-injection into the pronuclei of fertilized embryos which are then reimplanted in carrier females.
- transgenic animals is also possible by cloning by nuclear transfer followed by embryo transfer in recipient females.
- the subject of the present invention is also the milk produced by the transgenic rabbits in accordance with the invention, and the use of this milk as a raw material for the purification of recombinant human factor VII.
- Recombinant human factor VII can be purified from this milk by methods known per se, for example as described in US Pat. No. 6,268,487.
- the transgenesis vector used for cloning is derived from the plasmid pPolylII, having an ampicillin resistance gene as well as the bacterial origin of replication Col El.
- This transgenesis vector contains an expression cassette comprising: a dimer of the sequence of the 5 ⁇ S4 isolator of the chicken beta-globin gene (Genbank U78775) (Recillas-Targa et al, Proc Natl Acad ScL 5 : 6883-6888, 2002) upstream of the WAP promoter (whey acidic protein; Genbank X52564) of 6.3 kbp rabbit (Rival-Gervier et al, Transgenic Res., 6: 723-730, 2003), the first intron of the rabbit beta-globin gene (Genbank V00882) containing a transcription enhancer ; a second transcription amplifier (SUR 1.2.3) containing the 5 'UTR sequence of the SV40 early genes fused with the R region and the beginning of the U5 region of HT
- the FVII encoding DNA insert recovered by MlullNhel digestion from the intermediate vector, was inserted between the MwI and NheI sites of the expression cassette.
- the resulting transgenesis vector is shown in Figure 1. It contains an insert which is constituted in its 5 'to 3' orientation by i) the dimer of the 5 ⁇ S4 isolator sequence of the chicken beta-globin gene, ii) the 6.3 kbp rabbit WAP (whey acidic protein) promoter, iii) the intron containing the first transcription enhancer, iv) the second transcription enhancer, v) the human factor VII cDNA, vi) the third transcriptional amplifier; and vii) the transcription terminator.
- the colonies containing the recombinant vector are selected on the basis of their resistance to ampicillin, and then the presence of the insert is monitored by restriction fragment analysis, followed by sequencing.
- Transgenic rabbits were obtained by the conventional microinjection technique (Brinster et al, Proc Natl Acad Soi 82: 4438-4442, 1985).
- transgenesis vector containing the recombinant factor VII coding sequence was digested with the NotI restriction enzyme and the insert containing the transgene was isolated on agarose gel and then purified on ElutipD (Schleicher-Schuell,
- Microinjection of oocytes taken and implantation On the 4th day, 18-19h after the mating, the embryos are taken from the donor rabbits to carry out microinjection of DNA: the microinjection of DNA takes place just after samples (15-25 h after mating).
- the one-cell stage embryos are placed in a micro-drop of medium under an inverted microscope equipped with Normarsky objectives and micro-manipulators. Individual embryos are positioned and secured using a pipette.
- Intact unicellular embryos are then reimplanted under general anesthesia into the lumen of the oviducts of the synchronized recipient rabbits (10 embryos in each oviduct), using a surgical procedure (the oviducts are externalised by laparotomy). Parturition may occur naturally 29-31 days after embryo transfer. If necessary, oxytocin injection is initiated on the 31st day. The number of rabbits born in relation to the number of reimplanted embryos is of the order of 5 to 20%. EXAMPLE 3 SELECTION AND CHARACTERIZATION OF TRANSGENIC RABBITS
- the microinjected recombinant DNA integrates the genome randomly. Newborn rabbits (10 days) are tested for the presence of the transgene by an ear biopsy. The genomic DNA is extracted and PCR (Polymerase Chain Reaction) analysis is performed using primers specific for the recombinant insert.
- PCR Polymerase Chain Reaction
- the lineages of the FO founders were further characterized by i) analyzing the number of transgene copies integrated into their genome, and ii) determining the number of integration sites.
- the number of copies of the transgene integrated into the genome of each FO founder line was determined by quantitative PCR and Southern blotting. This number varies, depending on the line, from 1 copy per cell to more than 200 copies per cell.
- the number of integration sites has also been determined by Southern blotting. This number varies, depending on the line, from 1 to more than 5 sites per genome.
- Detection was performed by chemiluminescence (Amersham Biosciences) to determine the human factor VII concentration for each of the milks obtained. An expression level of factor VII varying between 90 ⁇ g / ml and 11 mg / ml was observed. The influence of the number of copies of the transgene on the level of expression has also been sought. The results are illustrated in Figure 2, and show a correlation between the transgene copy number and the expression level of factor VII in the milk of transgenic animals.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Veterinary Medicine (AREA)
- Biophysics (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Hematology (AREA)
- Physics & Mathematics (AREA)
- Animal Husbandry (AREA)
- Plant Pathology (AREA)
- Toxicology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Diabetes (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
Claims
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2011501259A JP2011517562A (ja) | 2008-03-25 | 2008-03-25 | ヒト第vii因子を産生するトランスジェニックウサギ |
EP08787842A EP2271206A1 (fr) | 2008-03-25 | 2008-03-25 | Lapins transgeniques producteurs de facteur vii humain |
CA2719459A CA2719459A1 (fr) | 2008-03-25 | 2008-03-25 | Lapins transgeniques producteurs de facteur vii humain |
PCT/FR2008/000396 WO2009118460A1 (fr) | 2008-03-25 | 2008-03-25 | Lapins transgeniques producteurs de facteur vii humain |
US12/934,124 US20110059510A1 (en) | 2008-03-25 | 2008-03-25 | Transgenic rabbits producing human factor vii |
BRPI0822557A BRPI0822557A2 (pt) | 2008-03-25 | 2008-03-25 | coelhos transgênicos produtores de fator vii humano |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
PCT/FR2008/000396 WO2009118460A1 (fr) | 2008-03-25 | 2008-03-25 | Lapins transgeniques producteurs de facteur vii humain |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2009118460A1 true WO2009118460A1 (fr) | 2009-10-01 |
Family
ID=40136350
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/FR2008/000396 WO2009118460A1 (fr) | 2008-03-25 | 2008-03-25 | Lapins transgeniques producteurs de facteur vii humain |
Country Status (6)
Country | Link |
---|---|
US (1) | US20110059510A1 (fr) |
EP (1) | EP2271206A1 (fr) |
JP (1) | JP2011517562A (fr) |
BR (1) | BRPI0822557A2 (fr) |
CA (1) | CA2719459A1 (fr) |
WO (1) | WO2009118460A1 (fr) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2687595B1 (fr) | 2012-07-19 | 2018-05-30 | Laboratoire Français du Fractionnement et des Biotechnologies | Procédé de purification de facteur VII transgénique |
FR3082427B1 (fr) | 2018-06-14 | 2020-09-25 | Lab Francais Du Fractionnement | Combinaison de facteur vii et d'un anticorps bispecifique anti-facteurs ix et x |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007138199A2 (fr) * | 2006-05-31 | 2007-12-06 | Lfb Biotechnologies | Composition de facteur vii recombinant ou transgenique, chaque molecule de facteur vii possedant deux sites de n-glycosylation a motifs glycanniques definis |
WO2008015339A2 (fr) * | 2006-08-01 | 2008-02-07 | Lfb Biotechnologies | Composition de facteur vii recombinant |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
ATE455552T1 (de) * | 1997-02-14 | 2010-02-15 | American Nat Red Cross | Expression des active human factor ix im brustdrüsengewebe transgener tiere |
-
2008
- 2008-03-25 JP JP2011501259A patent/JP2011517562A/ja active Pending
- 2008-03-25 WO PCT/FR2008/000396 patent/WO2009118460A1/fr active Application Filing
- 2008-03-25 US US12/934,124 patent/US20110059510A1/en not_active Abandoned
- 2008-03-25 EP EP08787842A patent/EP2271206A1/fr not_active Withdrawn
- 2008-03-25 BR BRPI0822557A patent/BRPI0822557A2/pt not_active IP Right Cessation
- 2008-03-25 CA CA2719459A patent/CA2719459A1/fr not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2007138199A2 (fr) * | 2006-05-31 | 2007-12-06 | Lfb Biotechnologies | Composition de facteur vii recombinant ou transgenique, chaque molecule de facteur vii possedant deux sites de n-glycosylation a motifs glycanniques definis |
WO2008015339A2 (fr) * | 2006-08-01 | 2008-02-07 | Lfb Biotechnologies | Composition de facteur vii recombinant |
Non-Patent Citations (3)
Title |
---|
ERIC SOLER ET AL: "Production of Two Vaccinating Recombinant Rotavirus Proteins in the Milk of Transgenic Rabbits", TRANSGENIC RESEARCH, KLUWER ACADEMIC PUBLISHERS-PLENUM PUBLISHERS, NE, vol. 14, no. 6, 1 December 2005 (2005-12-01), pages 833 - 844, XP019269494, ISSN: 1573-9368 * |
GALET C ET AL: "EXPRESSION OF A SINGLE BETAALPHA CHAIN PROTEIN OF EQUINE LH/CG IN MILK OF TRANSGENIC RABBITS AND ITS BIOLOGICAL ACTIVITY", MOLECULAR AND CELLULAR ENDOCRINOLOGY, ELSEVIER IRELAND LTD, IE, vol. 174, no. 1/02, 28 March 2001 (2001-03-28), pages 31 - 40, XP001015528, ISSN: 0303-7207 * |
See also references of EP2271206A1 * |
Also Published As
Publication number | Publication date |
---|---|
EP2271206A1 (fr) | 2011-01-12 |
JP2011517562A (ja) | 2011-06-16 |
CA2719459A1 (fr) | 2009-10-01 |
BRPI0822557A2 (pt) | 2019-09-24 |
US20110059510A1 (en) | 2011-03-10 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Houdebine | Production of pharmaceutical proteins by transgenic animals | |
US7888321B2 (en) | Production of high levels of transgenic factor IX without gene rescue, and its therapeutic uses | |
US7435869B2 (en) | Transgenic nonhuman mammals producing fibrinogen in milk and methods of producing fibrin | |
WO1998035689A1 (fr) | Expression du facteur ix humain actif, dans un tissu mammaire d'animal transgenique | |
JP2010017184A (ja) | ヒトの凝固第viii因子及びフォンビルブラント因子を発現するトランスジェニック動物 | |
CZ309194A3 (en) | Dna sequences used in production of recombinant bssl/cel in transgenic mammals who are not human beings and produced bssl/cel in suckling's food | |
US20070011752A1 (en) | Production of human proteins in transgenic animal saliva | |
EP0744891B1 (fr) | Fibrinogene transgenique | |
US20130131317A1 (en) | Expression of secreted human alpha-fetoprotein in transgenic animals | |
WO2002072023A2 (fr) | Production de niveaux eleves de facteur transgeniques viii avec stabilite etudiee, et ses applications therapeutiques | |
EP2271206A1 (fr) | Lapins transgeniques producteurs de facteur vii humain | |
US20110283375A1 (en) | Transgenic Prothrombin And Related Thrombin Precursors And Transgenics, Methods, Compositions, Uses And The Like Relating Thereto | |
JP2004500890A (ja) | 遺伝子導入によって作成された血小板由来増殖因子 | |
JPH10502816A (ja) | α−ラクトアルブミン遺伝子構造物 | |
US20040117862A1 (en) | Production of high levels of transgenic factor VII with engineered stability and its therapeutic uses | |
EP2555611B1 (fr) | Préparation de protéine plasmatique de transfert des phospholipides (pltp) humaine recombinante a partir du lait d'animaux transgéniques | |
WO2000024759A1 (fr) | Administration systemique de produits geniques par l'intermediaire la peau | |
WO2023019233A1 (fr) | Protéines de facteur ix recombinantes, leurs méthodes de fabrication et leurs méthodes d'utilisation chez des sujets non hémophiles | |
Velander et al. | TREATMENT OF HEMOPHILIA WITH HUMAN FACTORIX PRODUCED IN MAMIMARY TISSUE OF TRANSGENIC MAMMALS | |
Mohammadian et al. | Investigation of hFVIII Production in Mammary Glands of Transgenic Mice | |
Velander et al. | TRANSGENIC NONHUMAN MAMMALS PRODUCING FIBRINOGEN IN MILKAND METHODS OF PRODUCING FIBRIN | |
AU2002254193A1 (en) | Production of high levels of transgenic factor VIII with engineered stability, and its therapeutic uses | |
AU2002305043A1 (en) | Production of high levels of trangenic factor IX without gene rescue, and its therapeutic uses |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 08787842 Country of ref document: EP Kind code of ref document: A1 |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2011501259 Country of ref document: JP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2719459 Country of ref document: CA |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2008787842 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 12934124 Country of ref document: US |
|
ENP | Entry into the national phase |
Ref document number: PI0822557 Country of ref document: BR Kind code of ref document: A2 Effective date: 20100924 |