JP2008533980A - ジフテリア毒素生産のための発酵方法 - Google Patents
ジフテリア毒素生産のための発酵方法 Download PDFInfo
- Publication number
- JP2008533980A JP2008533980A JP2008502343A JP2008502343A JP2008533980A JP 2008533980 A JP2008533980 A JP 2008533980A JP 2008502343 A JP2008502343 A JP 2008502343A JP 2008502343 A JP2008502343 A JP 2008502343A JP 2008533980 A JP2008533980 A JP 2008533980A
- Authority
- JP
- Japan
- Prior art keywords
- diphtheria
- fermentation
- fermentation step
- culture
- kla
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000000855 fermentation Methods 0.000 title claims abstract description 111
- 230000004151 fermentation Effects 0.000 title claims abstract description 110
- 238000000034 method Methods 0.000 title claims abstract description 82
- 108010053187 Diphtheria Toxin Proteins 0.000 title claims description 47
- 102000016607 Diphtheria Toxin Human genes 0.000 title claims description 47
- 238000004519 manufacturing process Methods 0.000 title claims description 21
- 206010013023 diphtheria Diseases 0.000 claims abstract description 54
- 238000005273 aeration Methods 0.000 claims abstract description 44
- 238000013019 agitation Methods 0.000 claims abstract description 26
- 241000186216 Corynebacterium Species 0.000 claims abstract description 3
- 108010071134 CRM197 (non-toxic variant of diphtheria toxin) Proteins 0.000 claims description 57
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 53
- 108091007433 antigens Proteins 0.000 claims description 37
- 102000036639 antigens Human genes 0.000 claims description 37
- 239000002609 medium Substances 0.000 claims description 35
- 150000004676 glycans Chemical class 0.000 claims description 34
- 229920001282 polysaccharide Polymers 0.000 claims description 34
- 239000005017 polysaccharide Substances 0.000 claims description 34
- 239000000427 antigen Substances 0.000 claims description 33
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 26
- 239000001301 oxygen Substances 0.000 claims description 26
- 229910052760 oxygen Inorganic materials 0.000 claims description 26
- 239000012634 fragment Substances 0.000 claims description 24
- 229910052742 iron Inorganic materials 0.000 claims description 17
- 238000003756 stirring Methods 0.000 claims description 12
- 229920001542 oligosaccharide Polymers 0.000 claims description 11
- 150000002482 oligosaccharides Chemical class 0.000 claims description 11
- 239000008194 pharmaceutical composition Substances 0.000 claims description 11
- 230000001580 bacterial effect Effects 0.000 claims description 9
- 239000000523 sample Substances 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 8
- 239000002253 acid Substances 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 7
- 239000001963 growth medium Substances 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 5
- 239000003937 drug carrier Substances 0.000 claims description 5
- 208000015181 infectious disease Diseases 0.000 claims description 5
- 208000035143 Bacterial infection Diseases 0.000 claims description 4
- 239000002518 antifoaming agent Substances 0.000 claims description 4
- 208000022362 bacterial infectious disease Diseases 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 230000002265 prevention Effects 0.000 claims description 4
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 244000068988 Glycine max Species 0.000 claims description 2
- 235000010469 Glycine max Nutrition 0.000 claims description 2
- 229940041514 candida albicans extract Drugs 0.000 claims description 2
- 239000005018 casein Substances 0.000 claims 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 claims description 2
- 235000021240 caseins Nutrition 0.000 claims description 2
- 230000001268 conjugating effect Effects 0.000 claims description 2
- 159000000014 iron salts Chemical class 0.000 claims description 2
- 239000012138 yeast extract Substances 0.000 claims description 2
- 241000196324 Embryophyta Species 0.000 claims 1
- 231100000765 toxin Toxicity 0.000 description 21
- 239000000203 mixture Substances 0.000 description 19
- 108090000623 proteins and genes Proteins 0.000 description 18
- 239000003053 toxin Substances 0.000 description 18
- 108700012359 toxins Proteins 0.000 description 18
- 229960005486 vaccine Drugs 0.000 description 18
- 235000018102 proteins Nutrition 0.000 description 16
- 102000004169 proteins and genes Human genes 0.000 description 16
- 230000002163 immunogen Effects 0.000 description 15
- 229960003983 diphtheria toxoid Drugs 0.000 description 12
- 239000000499 gel Substances 0.000 description 10
- 230000035772 mutation Effects 0.000 description 10
- 241000894006 Bacteria Species 0.000 description 8
- 108010078791 Carrier Proteins Proteins 0.000 description 8
- 102000014914 Carrier Proteins Human genes 0.000 description 8
- 238000001471 micro-filtration Methods 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 201000005702 Pertussis Diseases 0.000 description 7
- 230000037430 deletion Effects 0.000 description 7
- 238000012217 deletion Methods 0.000 description 7
- 238000011081 inoculation Methods 0.000 description 7
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 7
- 229960000814 tetanus toxoid Drugs 0.000 description 7
- 241000606768 Haemophilus influenzae Species 0.000 description 6
- 101710183389 Pneumolysin Proteins 0.000 description 6
- 230000012010 growth Effects 0.000 description 6
- 239000012466 permeate Substances 0.000 description 6
- 239000000725 suspension Substances 0.000 description 6
- 230000015572 biosynthetic process Effects 0.000 description 5
- 238000011026 diafiltration Methods 0.000 description 5
- 238000011084 recovery Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 101710085938 Matrix protein Proteins 0.000 description 4
- 101710127721 Membrane protein Proteins 0.000 description 4
- 241000588650 Neisseria meningitidis Species 0.000 description 4
- 241000193998 Streptococcus pneumoniae Species 0.000 description 4
- 239000002671 adjuvant Substances 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000012228 culture supernatant Substances 0.000 description 4
- 239000007789 gas Substances 0.000 description 4
- 239000002054 inoculum Substances 0.000 description 4
- 229940031000 streptococcus pneumoniae Drugs 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 231100000419 toxicity Toxicity 0.000 description 4
- 230000001988 toxicity Effects 0.000 description 4
- -1 CRM228 Proteins 0.000 description 3
- 102100037840 Dehydrogenase/reductase SDR family member 2, mitochondrial Human genes 0.000 description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 3
- 241000588653 Neisseria Species 0.000 description 3
- 101710188053 Protein D Proteins 0.000 description 3
- 101710132893 Resolvase Proteins 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 230000005587 bubbling Effects 0.000 description 3
- 230000015556 catabolic process Effects 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 230000007423 decrease Effects 0.000 description 3
- 238000006731 degradation reaction Methods 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 229940047650 haemophilus influenzae Drugs 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 230000003053 immunization Effects 0.000 description 3
- 238000002649 immunization Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- 108090000765 processed proteins & peptides Proteins 0.000 description 3
- 238000011002 quantification Methods 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 241000712461 unidentified influenza virus Species 0.000 description 3
- WTLKTXIHIHFSGU-UHFFFAOYSA-N 2-nitrosoguanidine Chemical compound NC(N)=NN=O WTLKTXIHIHFSGU-UHFFFAOYSA-N 0.000 description 2
- QFVHZQCOUORWEI-UHFFFAOYSA-N 4-[(4-anilino-5-sulfonaphthalen-1-yl)diazenyl]-5-hydroxynaphthalene-2,7-disulfonic acid Chemical compound C=12C(O)=CC(S(O)(=O)=O)=CC2=CC(S(O)(=O)=O)=CC=1N=NC(C1=CC=CC(=C11)S(O)(=O)=O)=CC=C1NC1=CC=CC=C1 QFVHZQCOUORWEI-UHFFFAOYSA-N 0.000 description 2
- 108010077805 Bacterial Proteins Proteins 0.000 description 2
- 108010034055 CRM45 fragment of diphtheria toxin Proteins 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 101710099976 Photosystem I P700 chlorophyll a apoprotein A1 Proteins 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 241000191963 Staphylococcus epidermidis Species 0.000 description 2
- 241001505901 Streptococcus sp. 'group A' Species 0.000 description 2
- 206010043376 Tetanus Diseases 0.000 description 2
- 108010031133 Transferrin-Binding Protein A Proteins 0.000 description 2
- 108010031127 Transferrin-Binding Protein B Proteins 0.000 description 2
- IBVAQQYNSHJXBV-UHFFFAOYSA-N adipic acid dihydrazide Chemical compound NNC(=O)CCCCC(=O)NN IBVAQQYNSHJXBV-UHFFFAOYSA-N 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000002238 attenuated effect Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 239000004643 cyanate ester Substances 0.000 description 2
- 238000001739 density measurement Methods 0.000 description 2
- 229910001873 dinitrogen Inorganic materials 0.000 description 2
- 239000006260 foam Substances 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 108010037896 heparin-binding hemagglutinin Proteins 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 206010022000 influenza Diseases 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000007918 intramuscular administration Methods 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 238000010979 pH adjustment Methods 0.000 description 2
- 238000001243 protein synthesis Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000014616 translation Effects 0.000 description 2
- STCOOQWBFONSKY-UHFFFAOYSA-N tributyl phosphate Chemical compound CCCCOP(=O)(OCCCC)OCCCC STCOOQWBFONSKY-UHFFFAOYSA-N 0.000 description 2
- YYGNTYWPHWGJRM-UHFFFAOYSA-N (6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyltetracosa-2,6,10,14,18,22-hexaene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC=C(C)CCC=C(C)CCC=C(C)C YYGNTYWPHWGJRM-UHFFFAOYSA-N 0.000 description 1
- OTLLEIBWKHEHGU-UHFFFAOYSA-N 2-[5-[[5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy]-3,4-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3,5-dihydroxy-4-phosphonooxyhexanedioic acid Chemical compound C1=NC=2C(N)=NC=NC=2N1C(C(C1O)O)OC1COC1C(CO)OC(OC(C(O)C(OP(O)(O)=O)C(O)C(O)=O)C(O)=O)C(O)C1O OTLLEIBWKHEHGU-UHFFFAOYSA-N 0.000 description 1
- OMCVXIQHMVXMNN-DFWYDOINSA-N 2-aminoacetic acid;(2s)-2,5-diamino-5-oxopentanoic acid Chemical compound NCC(O)=O.OC(=O)[C@@H](N)CCC(N)=O OMCVXIQHMVXMNN-DFWYDOINSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 231100000699 Bacterial toxin Toxicity 0.000 description 1
- 241000588832 Bordetella pertussis Species 0.000 description 1
- 101000679617 Bordetella pertussis (strain Tohama I / ATCC BAA-589 / NCTC 13251) Pertussis toxin subunit 1 Proteins 0.000 description 1
- 108010053406 CRM 107 Proteins 0.000 description 1
- 101710163595 Chaperone protein DnaK Proteins 0.000 description 1
- 102000009016 Cholera Toxin Human genes 0.000 description 1
- 108010049048 Cholera Toxin Proteins 0.000 description 1
- 101710164918 Choline-binding protein Proteins 0.000 description 1
- 241000193449 Clostridium tetani Species 0.000 description 1
- 108010060123 Conjugate Vaccines Proteins 0.000 description 1
- XZMCDFZZKTWFGF-UHFFFAOYSA-N Cyanamide Chemical compound NC#N XZMCDFZZKTWFGF-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 101000874355 Escherichia coli (strain K12) Outer membrane protein assembly factor BamA Proteins 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 101100406392 Haemophilus influenzae (strain ATCC 51907 / DSM 11121 / KW20 / Rd) omp26 gene Proteins 0.000 description 1
- 101710178376 Heat shock 70 kDa protein Proteins 0.000 description 1
- 101710152018 Heat shock cognate 70 kDa protein Proteins 0.000 description 1
- 101710133291 Hemagglutinin-neuraminidase Proteins 0.000 description 1
- 241000700721 Hepatitis B virus Species 0.000 description 1
- 241000709721 Hepatovirus A Species 0.000 description 1
- 101000957351 Homo sapiens Myc-associated zinc finger protein Proteins 0.000 description 1
- 101000873843 Homo sapiens Sorting and assembly machinery component 50 homolog Proteins 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- 108090001030 Lipoproteins Proteins 0.000 description 1
- 102000004895 Lipoproteins Human genes 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 102100038750 Myc-associated zinc finger protein Human genes 0.000 description 1
- 241000186359 Mycobacterium Species 0.000 description 1
- QVLMCRFQGHWOPM-ZKWNWVNESA-N N-arachidonoyl vanillylamine Chemical compound CCCCC\C=C/C\C=C/C\C=C/C\C=C/CCCC(=O)NCC1=CC=C(O)C(OC)=C1 QVLMCRFQGHWOPM-ZKWNWVNESA-N 0.000 description 1
- 102100035181 Plastin-1 Human genes 0.000 description 1
- 101710110023 Putative adhesin Proteins 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 102100035853 Sorting and assembly machinery component 50 homolog Human genes 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- BHEOSNUKNHRBNM-UHFFFAOYSA-N Tetramethylsqualene Natural products CC(=C)C(C)CCC(=C)C(C)CCC(C)=CCCC=C(C)CCC(C)C(=C)CCC(C)C(C)=C BHEOSNUKNHRBNM-UHFFFAOYSA-N 0.000 description 1
- UZQJVUCHXGYFLQ-AYDHOLPZSA-N [(2s,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-4-[(2r,3r,4s,5r,6r)-4-[(2s,3r,4s,5r,6r)-3,5-dihydroxy-6-(hydroxymethyl)-4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-3,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3,5-dihydroxy-6-(hy Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O)O[C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O)O[C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2[C@@]1(C=O)C)C)(C)CC(O)[C@]1(CCC(CC14)(C)C)C(=O)O[C@H]1[C@@H]([C@@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O[C@H]4[C@@H]([C@@H](O[C@H]5[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O5)O)[C@H](O)[C@@H](CO)O4)O)[C@H](O)[C@@H](CO)O3)O)[C@H](O)[C@@H](CO)O2)O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O UZQJVUCHXGYFLQ-AYDHOLPZSA-N 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000004103 aerobic respiration Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- AZDRQVAHHNSJOQ-UHFFFAOYSA-N alumane Chemical class [AlH3] AZDRQVAHHNSJOQ-UHFFFAOYSA-N 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000000688 bacterial toxin Substances 0.000 description 1
- 229960001212 bacterial vaccine Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000021028 berry Nutrition 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- OWMVSZAMULFTJU-UHFFFAOYSA-N bis-tris Chemical compound OCCN(CCO)C(CO)(CO)CO OWMVSZAMULFTJU-UHFFFAOYSA-N 0.000 description 1
- 229920001400 block copolymer Polymers 0.000 description 1
- 150000001718 carbodiimides Chemical class 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000019365 chlortetracycline Nutrition 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 229940001442 combination vaccine Drugs 0.000 description 1
- 239000011365 complex material Substances 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 229940031670 conjugate vaccine Drugs 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 239000003431 cross linking reagent Substances 0.000 description 1
- 101150059761 ctrA gene Proteins 0.000 description 1
- ATDGTVJJHBUTRL-UHFFFAOYSA-N cyanogen bromide Chemical compound BrC#N ATDGTVJJHBUTRL-UHFFFAOYSA-N 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000007872 degassing Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 239000010432 diamond Substances 0.000 description 1
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N dodecahydrosqualene Natural products CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000002095 exotoxin Substances 0.000 description 1
- 231100000776 exotoxin Toxicity 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 229930195712 glutamate Natural products 0.000 description 1
- 102000006602 glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 229940045808 haemophilus influenzae type b Drugs 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- SYECJBOWSGTPLU-UHFFFAOYSA-N hexane-1,1-diamine Chemical compound CCCCCC(N)N SYECJBOWSGTPLU-UHFFFAOYSA-N 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000002297 mitogenic effect Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229940035032 monophosphoryl lipid a Drugs 0.000 description 1
- 125000001446 muramyl group Chemical group N[C@@H](C=O)[C@@H](O[C@@H](C(=O)*)C)[C@H](O)[C@H](O)CO 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 238000006213 oxygenation reaction Methods 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- 108010021711 pertactin Proteins 0.000 description 1
- 108010049148 plastin Proteins 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 235000004252 protein component Nutrition 0.000 description 1
- 101150054232 pyrG gene Proteins 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 239000000376 reactant Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000006722 reduction reaction Methods 0.000 description 1
- 238000006268 reductive amination reaction Methods 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 239000012465 retentate Substances 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 229940031439 squalene Drugs 0.000 description 1
- TUHBEKDERLKLEC-UHFFFAOYSA-N squalene Natural products CC(=CCCC(=CCCC(=CCCC=C(/C)CCC=C(/C)CC=C(C)C)C)C)C TUHBEKDERLKLEC-UHFFFAOYSA-N 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 238000011287 therapeutic dose Methods 0.000 description 1
- 231100000033 toxigenic Toxicity 0.000 description 1
- 230000001551 toxigenic effect Effects 0.000 description 1
- 238000013022 venting Methods 0.000 description 1
- 210000003501 vero cell Anatomy 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 239000000277 virosome Substances 0.000 description 1
- 230000001018 virulence Effects 0.000 description 1
- 230000003313 weakening effect Effects 0.000 description 1
Images
Classifications
-
- 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
- C12N9/10—Transferases (2.)
- C12N9/1048—Glycosyltransferases (2.4)
- C12N9/1077—Pentosyltransferases (2.4.2)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
- C07K14/34—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Corynebacterium (G)
-
- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
- C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
- C12N15/77—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Corynebacterium; for Brevibacterium
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
Description
ジフテリアトキソイドと、毒性が減弱された突然変異体型であるCRM197は、ジフテリア菌に対する免疫を提供する多数のワクチンの成分である。百日咳(Bordetella pertussis)、破傷風菌(Clostridium tetani)、ジフテリア菌、および随意にB型肝炎ウイルスおよび/またはヘモフィルス・インフルエンザ(Haemophilus influenzae)タイプbを予防することができる数種の混合ワクチンが知られている(例えば、WO 93/24148およびWO 97/00697、WO 02/055105を参照)。
時間に対してln (100−pO2)をプロットすることにより、直線の勾配(または角度係数)は−KLaである。
発酵ステップのKLaを測定するために、発酵槽に所望の量の水を充填し、発酵パラメーター(例えば、温度、圧力、攪拌および通気)を適用し、この系が定常状態に達するまで放置した。pO2プローブを100%で較正した。
CRM197(交差反応物質)の調製に用いる細菌は、A. Pappenheimerの方法(Nature New Biol. 233:8-11、1971)に従うニトロソグアニジン処理により取得したジフテリア菌の突然変異株である。これは、無毒化ジフテリア毒素(52アミノ酸:グリシンのグルタミン酸への突然変異)を発現する。これは、ATCCから取得し、これを39255と称する。発酵方法の概略を図2に示す。
前述した発酵のプロトコルを用いて、150Lスケールで2つの発酵を実施した(CDT199およびCDT206)。前培養および培養の条件を表2および3に示し、CRM197の収量を表4に示す。図3は、前培養増殖の典型的動力学のグラフを示す。O.D.(650 nm)が4.0〜6.0の間であるとき、培養は、明らかに指数増殖期にあり、pHはわずかしか変化しなかった。
CDT206の精密濾過のためのデータを示す。
低い通気条件下でのCRM197の発現レベルは、全発酵を通じて、pO2を5%以上に維持した高い通気条件下で達成されたものより一般に2〜4倍高かった。
あらゆる分解バンドを検出することができるように(クリッピング後、それぞれ35および23 kDの2サブユニットを検出することができる)、SDS-PAGEゲルを用いて還元条件下で培養物上清を泳動した(図4)。次に、これらをクーマシーブルーで染色した。
前記2つの発酵から得たサンプルのクーマシー染色ゲルを図4A(CDT199)および4B(CDT206)に示す。CRM197は予想した分子量(理論MW:58.4 kD)に現れた。発酵の終点のシグナル後に取得したサンプルにはパターンの変化が認められなかった(図4A、レーン9および10)ことから、CRM197は分解されていなかった。図4Aのレーン9および11は同等量のCRM197を示すことから、CRM197の量は精密濾過および最終濾過の影響を受けなかった。
20リットル発酵槽を用いた以外は、実施例2に記載したものと類似の方法を用いてジフテリア菌を発酵した。培養物を300 rpmで攪拌し、気流を3リットル/分に設定した。発酵を通してpHはほぼ中性に留まっていたため、その間、酸または塩基を添加する必要はなかった。
一定KLaの条件下でジフテリア菌を増殖させる発酵方法は、他のサイズおよび様々な設計の発酵槽での使用のために適合させることができる。表5に記載した発酵槽サイズ、気流および攪拌速度の条件に従い、優れた収量のCRM197生産が達成された。様々な設計の発酵槽において、3つの150L規模の発酵を実施した。
実施例3に記載した方法に従い、発酵の大部分を通してpO2が低くなるようにする、あるいは、5%pO2の一定設定値で、ジフテリア菌株ATCC39255の一連の発酵を20リットル規模で実施した。存在するFe3+の量は、Fe3+の添加なし、250 ppbのFe3+添加、500 ppbのFe3+添加および500 ppbのFe2+添加の間で様々であった。発酵終了時のCRM197の収量をSDS-PAGEゲルの密度測定により測定した。この方法は、ELISAにより達成したものより約20%低い結果をもたらす傾向がある。
CRM197の発現に影響しない鉄濃度の範囲をマイクロプレートにおいて決定した。マイクロプレートは、本発明の発酵方法に存在する制限された通気条件をシミュレートするものである。
表7は、マイクロタイターウェルにおいて制限された通気条件下で増殖させたジフテリア菌について、様々な鉄濃度でのCRM197の発現を示す。CRM197発現は、Fe3+による抑制に対する感受性が低く、1ppmまたは2ppmのFe3+の添加には有意に影響されなかった。3ppmの濃度でのみ、CRM197発現の有意な低下が起こった。このレベルのFe3+でも、CRM197の発現はまだ、Fe3+を添加しないで達成された発現の79%であった。
Claims (35)
- 均質な培養物を維持するのに十分な攪拌と、培養物内のpO2が、発酵ステップの大部分の間、4%以下になるような制限された通気の条件下で、ジフテリア菌(Corynebacterium diphtheria)株を発酵槽中の培地で増殖させる発酵ステップを含む、発酵方法。
- 前記pO2が、発酵ステップの大部分の間、0に近似する、請求項1に記載の発酵方法。
- 酸素の迅速な消費によりpO2が4%以下になるのに十分な密度までジフテリア菌が増殖した時点から、発酵ステップが終了するまで、4%以下のpO2を維持する、請求項1または2に記載の方法。
- 発酵ステップを一定のKLaで実施する、請求項1〜3のいずれか一項に記載の方法。
- 発酵ステップを一定の攪拌速度および通気速度で実施する、請求項1〜4のいずれか一項に記載の方法。
- 発酵ステップを可変KLa条件下で実施する、請求項1〜3のいずれか一項に記載の方法。
- 発酵ステップを10〜50h-1のKLaで実施する、請求項1〜6のいずれか一項に記載の方法。
- 10〜30リットル発酵槽において10〜30h-1のKLaで発酵ステップを実施する、請求項1〜7のいずれか一項に記載の方法。
- 100〜250リットル発酵槽において30〜60h-1のKLaで発酵ステップを実施する、請求項1〜7のいずれか一項に記載の方法。
- 250〜800リットル発酵槽において60〜150h-1のKLaで発酵ステップを実施する、請求項1〜6のいずれか一項に記載の方法。
- 1〜5L/分の圧縮空気の気流と200〜400rpmの攪拌速度で発酵ステップを実施する、請求項8に記載の方法。
- 15〜25L/分の圧縮空気の気流と150〜250rpmの攪拌速度で発酵ステップを実施する、請求項9に記載の方法。
- 培地がCY、SOCまたは類似の培地であり、通気の程度により発酵槽内のpHを7.0〜7.8に保持するが、その際、酸または塩基を添加する必要がない、請求項1〜12のいずれか一項に記載の方法。
- 前記ジフテリア菌株がジフテリア毒素またはその突然変異体、特にCRM197を産生する、請求項1〜13のいずれか一項に記載の方法。
- 前記ジフテリア菌株がATCC39255である、請求項1〜14のいずれか一項に記載の方法。
- 前記方法が、培地中の鉄塩の存在に十分耐性であり、それにより、使用前に、鉄を除去するための培地処理を必要としない、請求項1〜15のいずれか一項に記載の方法。
- 前記培地が10〜4,000 ppbの鉄を含む、請求項1〜16のいずれか一項に記載の方法。
- 前記培地が100〜3,000 ppbまたは1,700〜3,000 ppbの鉄を含む、請求項17に記載の方法。
- 前記培地が5〜20g/Lのカザミノ酸またはカゼイン加水分解物を含む、請求項1〜18のいずれか一項に記載の方法。
- 前記培地が5〜20g/Lの大豆ペプトンまたはその他の植物ペプトンを含む、請求項1〜18のいずれか一項に記載の方法。
- 前記培地が10〜30g/Lの酵母抽出物を含む、請求項1〜20のいずれか一項に記載の方法。
- 発酵を25〜40℃の温度で実施する、請求項1〜21のいずれか一項に記載の方法。
- 消泡剤を発酵槽に添加する、請求項1〜22のいずれか一項に記載の方法。
- 気泡プローブまたは機械的破泡機を発酵槽に用いる、請求項1〜23のいずれか一項に記載の方法。
- ジフテリア菌由来の抗原の調製物を製造する方法であって、請求項1〜24のいずれか一項に記載の発酵方法を実施するステップと、培養物からジフテリア菌由来の抗原を単離するステップを含む、上記方法。
- 前記ジフテリア菌株由来の抗原がジフテリア毒素またはそのフラグメントもしくは突然変異体である、請求項25に記載の方法。
- 前記抗原がCRM197である、請求項26に記載の方法。
- 1以上の追加の抗原をジフテリア菌株由来の抗原に付加するステップを含む、請求項25〜27のいずれか一項に記載の方法。
- ジフテリア毒素またはそのフラグメントもしくは突然変異体を1以上の追加の抗原とコンジュゲートさせるステップをさらに含む、請求項28に記載の方法。
- 1以上の追加の抗原が細菌多糖またはオリゴ糖を含む、請求項29に記載の方法。
- 請求項25〜30のいずれか一項に記載の方法により作製した(コンジュゲートされたまたはコンジュゲートされない)抗原またはジフテリア毒素もしくは突然変異体と、薬学的に許容される担体を混合するステップを含む、医薬組成物の製造方法。
- 細菌症の治療または予防のための医薬の製造における、請求項25〜31のいずれか一項に記載の方法により作製した抗原、ジフテリア毒素またはその突然変異体の使用。
- 前記細菌症がジフテリア菌症である、請求項32に記載の使用。
- 請求項31に記載の方法により製造した医薬組成物を患者に投与することを含む、細菌感染症の予防または治療方法。
- 前記細菌感染症がジフテリア菌感染症である、請求項34に記載の方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GB0505996.9 | 2005-03-23 | ||
GBGB0505996.9A GB0505996D0 (en) | 2005-03-23 | 2005-03-23 | Fermentation process |
PCT/EP2006/002835 WO2006100108A1 (en) | 2005-03-23 | 2006-03-21 | Fermentation process for the production of diphtheria toxin |
Publications (3)
Publication Number | Publication Date |
---|---|
JP2008533980A true JP2008533980A (ja) | 2008-08-28 |
JP2008533980A5 JP2008533980A5 (ja) | 2009-04-16 |
JP4917087B2 JP4917087B2 (ja) | 2012-04-18 |
Family
ID=34531759
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2008502343A Active JP4917087B2 (ja) | 2005-03-23 | 2006-03-21 | ジフテリア毒素生産のための発酵方法 |
Country Status (14)
Country | Link |
---|---|
US (2) | US20080193475A1 (ja) |
EP (2) | EP2174950B1 (ja) |
JP (1) | JP4917087B2 (ja) |
KR (1) | KR101341121B1 (ja) |
CN (1) | CN101180313B (ja) |
AT (2) | ATE540053T1 (ja) |
AU (1) | AU2006226542B2 (ja) |
BR (1) | BRPI0609692C1 (ja) |
CA (1) | CA2602143C (ja) |
DE (1) | DE602006010676D1 (ja) |
ES (2) | ES2378193T3 (ja) |
GB (1) | GB0505996D0 (ja) |
RU (1) | RU2394914C2 (ja) |
WO (1) | WO2006100108A1 (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20200010218A (ko) * | 2017-04-22 | 2020-01-30 | 바이오로지칼 이 리미티드 | Crm197의 높은 수준의 생산을 위한 개선된 방법 |
Families Citing this family (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
BRPI0916365A2 (pt) * | 2008-07-21 | 2018-05-02 | Brigham & Womens Hospital Inc | métodos e composições relacionados aos oligossacarídeos de glicosina beta-1,6 sintéticos |
IT1398927B1 (it) | 2009-06-25 | 2013-03-28 | Consorzio Interuniversitario Per Lo Sviluppo Dei Sistemi A Grande Interfase Csgi | Espressione batterica di un gene artificiale per la produzione di crm197 e derivati. |
CN104379170B (zh) | 2012-06-20 | 2017-05-03 | 国立大学法人东京大学 | 粘膜免疫刺激剂及hpv感染症治疗用经口药物组合物 |
CN103627758B (zh) * | 2012-08-28 | 2018-04-27 | 天士力医药集团股份有限公司 | 一种crm197蛋白的生产方法 |
US20140193451A1 (en) * | 2012-10-17 | 2014-07-10 | Glaxosmithkline Biologicals Sa | Immunogenic composition |
AR095425A1 (es) | 2013-03-15 | 2015-10-14 | Glaxosmithkline Biologicals Sa | Vacuna, uso y procedimiento para prevenir una infección por picornavirus |
US11951165B2 (en) | 2016-12-30 | 2024-04-09 | Vaxcyte, Inc. | Conjugated vaccine carrier proteins |
WO2018156465A1 (en) * | 2017-02-24 | 2018-08-30 | Merck Sharp & Dohme Corp. | Polysaccharide-protein conjugates utilizing diphtheria toxin fragment b as a carrier |
EP3699263A1 (en) | 2019-02-22 | 2020-08-26 | GlaxoSmithKline Biologicals S.A. | Fermentation process |
CN110452838B (zh) * | 2019-07-18 | 2020-12-29 | 艾美卫信生物药业(浙江)有限公司 | 一种crm197菌种培养基、配制方法及发酵培养方法 |
WO2021176409A1 (en) | 2020-03-05 | 2021-09-10 | Sanofi Healthcare India Private Limited | Preservative combination for vaccine composition |
CN114806973B (zh) * | 2022-06-08 | 2024-02-23 | 艾美坚持生物制药有限公司 | 一种提高crm197蛋白产量的生长因子及其制备方法和应用 |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH07300427A (ja) * | 1992-10-27 | 1995-11-14 | American Cyanamid Co | 各ワクチン成分の免疫原性を増強した併用小児ワクチン |
Family Cites Families (47)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4235877A (en) | 1979-06-27 | 1980-11-25 | Merck & Co., Inc. | Liposome particle containing viral or bacterial antigenic subunit |
DE3071552D1 (en) | 1979-09-21 | 1986-05-22 | Hitachi Ltd | Semiconductor switch |
US4673574A (en) | 1981-08-31 | 1987-06-16 | Anderson Porter W | Immunogenic conjugates |
CH660375A5 (it) | 1983-02-08 | 1987-04-15 | Sclavo Spa | Procedimento per la produzione di proteine correlate alla tossina difterica. |
US4695624A (en) | 1984-05-10 | 1987-09-22 | Merck & Co., Inc. | Covalently-modified polyanionic bacterial polysaccharides, stable covalent conjugates of such polysaccharides and immunogenic proteins with bigeneric spacers, and methods of preparing such polysaccharides and conjugates and of confirming covalency |
US4709017A (en) | 1985-06-07 | 1987-11-24 | President And Fellows Of Harvard College | Modified toxic vaccines |
IT1187753B (it) | 1985-07-05 | 1987-12-23 | Sclavo Spa | Coniugati glicoproteici ad attivita' immunogenica trivalente |
US4950740A (en) | 1987-03-17 | 1990-08-21 | Cetus Corporation | Recombinant diphtheria vaccines |
JP3237842B2 (ja) | 1988-12-16 | 2001-12-10 | オランダ国 | ニューモリシンミュータント及びそれから製造されたニューモコッカルワクチン |
DE69132581T3 (de) | 1990-03-02 | 2008-07-17 | Boston Medical Center Corp., Boston | Verbesserte chimäre toxine |
DE69133276T2 (de) | 1990-07-16 | 2003-11-27 | Univ North Carolina | Mit der familie der hämolysin-toxine verwandte antigene eisen-ubnterdrückende proteine des n. meningitis |
CA2087958C (en) | 1990-08-23 | 2007-05-15 | P. Frederick Sparling | Transferrin binding proteins from neisseria gonorrhoeae and neisseria meningitidis |
US5912336A (en) | 1990-08-23 | 1999-06-15 | University Of North Carolina At Chapel Hill | Isolated nucleic acid molecules encoding transferrin binding proteins from Neisseria gonorrhoeae and Neisseria meningitidis |
US5153312A (en) | 1990-09-28 | 1992-10-06 | American Cyanamid Company | Oligosaccharide conjugate vaccines |
US6592876B1 (en) | 1993-04-20 | 2003-07-15 | Uab Research Foundation | Pneumococcal genes, portions thereof, expression products therefrom, and uses of such genes, portions and products |
US5476929A (en) | 1991-02-15 | 1995-12-19 | Uab Research Foundation | Structural gene of pneumococcal protein |
FR2682041B1 (fr) | 1991-10-03 | 1994-01-14 | Pasteur Merieux Serums Vaccins | Vaccin contre les infections a neisseria meningitidis. |
ATE245446T1 (de) | 1992-02-11 | 2003-08-15 | Jackson H M Found Military Med | Dualer träger für immunogene konstrukte |
JP3506431B2 (ja) | 1992-05-06 | 2004-03-15 | プレジデント アンド フェローズ オブ ハーバード カレッジ | ジフテリア毒素受容体結合領域 |
NZ253065A (en) | 1992-05-23 | 1996-10-28 | Smithkline Beecham Biolog | Combination vaccines comprising hepatitis b surface antigens and other antigens wherein aluminium phosphate adjuvant is used to adsorb the hepatitis antigen |
AU671649B2 (en) * | 1992-06-18 | 1996-09-05 | President And Fellows Of Harvard College | Diphtheria toxin vaccines |
FR2692592B1 (fr) | 1992-06-19 | 1995-03-31 | Pasteur Merieux Serums Vacc | Fragments d'ADN codant pour les sous-unités du récepteur de la transferrine de Neisseria meningitidis et procédés les exprimant. |
DE69434079T2 (de) | 1993-03-05 | 2005-02-24 | Wyeth Holdings Corp. | Plasmid zur Herstellung von CRM-Protein und Diphtherie-Toxin |
WO1994026304A1 (en) | 1993-05-18 | 1994-11-24 | Ohio State Research Foundation | Otitis media vaccine |
US5917017A (en) * | 1994-06-08 | 1999-06-29 | President And Fellows Of Harvard College | Diphtheria toxin vaccines bearing a mutated R domain |
US6455673B1 (en) | 1994-06-08 | 2002-09-24 | President And Fellows Of Harvard College | Multi-mutant diphtheria toxin vaccines |
US5565204A (en) | 1994-08-24 | 1996-10-15 | American Cyanamid Company | Pneumococcal polysaccharide-recombinant pneumolysin conjugate vaccines for immunization against pneumococcal infections |
IL117483A (en) | 1995-03-17 | 2008-03-20 | Bernard Brodeur | MENINGITIDIS NEISSERIA shell protein is resistant to proteinase K. |
US6265567B1 (en) | 1995-04-07 | 2001-07-24 | University Of North Carolina At Chapel Hill | Isolated FrpB nucleic acid molecule |
US5843464A (en) | 1995-06-02 | 1998-12-01 | The Ohio State University | Synthetic chimeric fimbrin peptides |
JPH11507214A (ja) | 1995-06-07 | 1999-06-29 | バイオケム ヴァシーンズ インク. | Hsp70ファミリーに属する連鎖球菌の熱ショック蛋白質メンバー |
SI1082965T1 (sl) | 1995-06-23 | 2009-08-31 | Glaxosmithkline Biolog Sa | Sestavek cepiva, ki vsebuje antigen konjugata polisaharida, adsorbiranega na aluminijevem fosfatu |
EP0912608B1 (en) | 1996-05-01 | 2006-04-19 | The Rockefeller University | Choline binding proteins for anti-pneumococcal vaccines |
FR2751000B1 (fr) | 1996-07-12 | 1998-10-30 | Inst Nat Sante Rech Med | Adn specifiques des bacteries de l'espece neisseria meningitidis, leurs procedes d'obtention et leurs applications biologiques |
US5882871A (en) | 1996-09-24 | 1999-03-16 | Smithkline Beecham Corporation | Saliva binding protein |
US5882896A (en) | 1996-09-24 | 1999-03-16 | Smithkline Beecham Corporation | M protein |
EP1770164B1 (en) | 1996-10-31 | 2010-09-01 | Human Genome Sciences, Inc. | Streptococcus pneumoniae antigens and vaccines |
KR100619350B1 (ko) | 1997-07-21 | 2006-09-05 | 박스터 헬쓰케어 에스.에이. | 변형 면역성 뉴멀리신 백신조성물 |
GB9717953D0 (en) | 1997-08-22 | 1997-10-29 | Smithkline Beecham Biolog | Vaccine |
GB9726398D0 (en) | 1997-12-12 | 1998-02-11 | Isis Innovation | Polypeptide and coding sequences |
CN1200731C (zh) | 1998-04-07 | 2005-05-11 | 免疫医疗公司 | 用作疫苗的肺炎球菌胆碱结合蛋白衍生物 |
NZ532665A (en) | 1998-05-01 | 2005-11-25 | Inst Genomic Research | Neisseria meningitidis antigens and compositions |
DK1535928T3 (da) | 1998-10-22 | 2008-10-20 | Univ Montana | Vaccinesammensætninger indeholdende Omp85-proteiner af Neisseria gonorrhoeae og Neisseria meningitidis |
GB9904582D0 (en) | 1999-02-26 | 1999-04-21 | Nycomed Imaging As | Process |
KR100401423B1 (ko) | 2001-01-10 | 2003-10-17 | 주식회사 엘지생명과학 | 혼합 백신의 제조 방법 |
WO2006042542A2 (en) * | 2004-10-19 | 2006-04-27 | Statens Serum Institut | Production of tetanus, diphtheria, and pertussis toxins and toxoids using fermentation media containing no components of animal or soy origin |
US9473463B2 (en) | 2014-07-29 | 2016-10-18 | Combined Conditional Access Development & Support, LLC | Control word and associated entitlement control message caching and reuse |
-
2005
- 2005-03-23 GB GBGB0505996.9A patent/GB0505996D0/en not_active Ceased
-
2006
- 2006-03-21 CN CN2006800177931A patent/CN101180313B/zh active Active
- 2006-03-21 AT AT09176267T patent/ATE540053T1/de active
- 2006-03-21 BR BRPI0609692A patent/BRPI0609692C1/pt active IP Right Grant
- 2006-03-21 ES ES09176267T patent/ES2378193T3/es active Active
- 2006-03-21 ES ES06723804T patent/ES2335133T3/es active Active
- 2006-03-21 AT AT06723804T patent/ATE449788T1/de not_active IP Right Cessation
- 2006-03-21 EP EP09176267A patent/EP2174950B1/en active Active
- 2006-03-21 DE DE602006010676T patent/DE602006010676D1/de active Active
- 2006-03-21 KR KR1020077024097A patent/KR101341121B1/ko not_active IP Right Cessation
- 2006-03-21 RU RU2007135122/13A patent/RU2394914C2/ru not_active IP Right Cessation
- 2006-03-21 WO PCT/EP2006/002835 patent/WO2006100108A1/en active Application Filing
- 2006-03-21 CA CA2602143A patent/CA2602143C/en active Active
- 2006-03-21 US US11/909,100 patent/US20080193475A1/en not_active Abandoned
- 2006-03-21 JP JP2008502343A patent/JP4917087B2/ja active Active
- 2006-03-21 AU AU2006226542A patent/AU2006226542B2/en not_active Ceased
- 2006-03-21 EP EP06723804A patent/EP1861420B1/en active Active
-
2014
- 2014-03-14 US US14/210,626 patent/US20140242635A1/en not_active Abandoned
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH07300427A (ja) * | 1992-10-27 | 1995-11-14 | American Cyanamid Co | 各ワクチン成分の免疫原性を増強した併用小児ワクチン |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20200010218A (ko) * | 2017-04-22 | 2020-01-30 | 바이오로지칼 이 리미티드 | Crm197의 높은 수준의 생산을 위한 개선된 방법 |
JP2020517274A (ja) * | 2017-04-22 | 2020-06-18 | バイオロジカル イー リミテッド | Crm197の高レベル製造のための改良法 |
JP7161493B2 (ja) | 2017-04-22 | 2022-10-26 | バイオロジカル イー リミテッド | Crm197の高レベル製造のための改良法 |
KR102637436B1 (ko) | 2017-04-22 | 2024-02-15 | 바이오로지칼 이 리미티드 | Crm197의 높은 수준의 생산을 위한 개선된 방법 |
Also Published As
Publication number | Publication date |
---|---|
ATE449788T1 (de) | 2009-12-15 |
CA2602143A1 (en) | 2006-09-28 |
BRPI0609692A2 (pt) | 2010-04-20 |
RU2394914C2 (ru) | 2010-07-20 |
CN101180313A (zh) | 2008-05-14 |
WO2006100108A1 (en) | 2006-09-28 |
CN101180313B (zh) | 2012-10-03 |
EP1861420A1 (en) | 2007-12-05 |
BRPI0609692B1 (pt) | 2019-02-12 |
ATE540053T1 (de) | 2012-01-15 |
BRPI0609692B8 (pt) | 2020-01-21 |
GB0505996D0 (en) | 2005-04-27 |
AU2006226542A1 (en) | 2006-09-28 |
KR101341121B1 (ko) | 2013-12-12 |
RU2007135122A (ru) | 2009-04-27 |
JP4917087B2 (ja) | 2012-04-18 |
AU2006226542B2 (en) | 2012-02-09 |
EP2174950B1 (en) | 2012-01-04 |
CA2602143C (en) | 2016-08-09 |
ES2378193T3 (es) | 2012-04-09 |
DE602006010676D1 (de) | 2010-01-07 |
ES2335133T3 (es) | 2010-03-22 |
EP1861420B1 (en) | 2009-11-25 |
US20080193475A1 (en) | 2008-08-14 |
KR20070116132A (ko) | 2007-12-06 |
US20140242635A1 (en) | 2014-08-28 |
BRPI0609692C1 (pt) | 2021-05-25 |
EP2174950A1 (en) | 2010-04-14 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP4917087B2 (ja) | ジフテリア毒素生産のための発酵方法 | |
EP1849860B1 (en) | Process for preparing an immunogenic factor of Corynebacterium diphtheriae using a culture medium with yeast extract as aminoacid source and no protein complexes of animal origin | |
JP4827196B2 (ja) | 髄膜炎菌類タンパク質nmb1870のドメインおよびエピトープ | |
CN101932698A (zh) | 培养链球菌的发酵工艺以及从中提取cps的纯化工艺 | |
EP1951886A2 (en) | Fed batch culture methods for streptococci | |
JP6582020B2 (ja) | 発酵方法 | |
US9284526B2 (en) | Culture medium with yeast or soy bean extract as amino acid source and no protein complexes of animal origin | |
JP7394065B2 (ja) | ボルデテラ属種を培養する方法 | |
AU720156B2 (en) | Live attenuated RTX-producing bacteria of the family pasteurellaceae | |
US11970690B2 (en) | Methods of cultivating Bordetella species |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20090225 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20090225 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20110906 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20111206 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20120110 |
|
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20120125 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20150203 Year of fee payment: 3 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 4917087 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |