JP2021522785A - 抗体発現を最適化するための方法 - Google Patents
抗体発現を最適化するための方法 Download PDFInfo
- Publication number
- JP2021522785A JP2021522785A JP2020560954A JP2020560954A JP2021522785A JP 2021522785 A JP2021522785 A JP 2021522785A JP 2020560954 A JP2020560954 A JP 2020560954A JP 2020560954 A JP2020560954 A JP 2020560954A JP 2021522785 A JP2021522785 A JP 2021522785A
- Authority
- JP
- Japan
- Prior art keywords
- strain
- protein
- cell
- phenylalanine
- plasmid
- 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.)
- Pending
Links
- 230000014509 gene expression Effects 0.000 title claims abstract description 99
- 238000000034 method Methods 0.000 title claims abstract description 86
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 318
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 269
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 106
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 91
- 229920001184 polypeptide Polymers 0.000 claims abstract description 80
- 239000012634 fragment Substances 0.000 claims abstract description 23
- 235000018102 proteins Nutrition 0.000 claims description 250
- 235000001014 amino acid Nutrition 0.000 claims description 232
- 229940024606 amino acid Drugs 0.000 claims description 230
- 210000004027 cell Anatomy 0.000 claims description 227
- 150000001413 amino acids Chemical class 0.000 claims description 221
- 239000013612 plasmid Substances 0.000 claims description 167
- 150000007523 nucleic acids Chemical group 0.000 claims description 104
- 108010006519 Molecular Chaperones Proteins 0.000 claims description 88
- 239000013598 vector Substances 0.000 claims description 84
- 101150048892 parB gene Proteins 0.000 claims description 73
- 101100278012 Escherichia coli (strain K12) dnaG gene Proteins 0.000 claims description 72
- 101100165173 Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720) basS gene Proteins 0.000 claims description 72
- 101100421924 Thermus thermophilus (strain ATCC BAA-163 / DSM 7039 / HB27) spo0C gene Proteins 0.000 claims description 72
- 101150004979 flmA gene Proteins 0.000 claims description 72
- -1 CD-70 Proteins 0.000 claims description 49
- 102000008394 Immunoglobulin Fragments Human genes 0.000 claims description 44
- 108010021625 Immunoglobulin Fragments Proteins 0.000 claims description 44
- 102000005431 Molecular Chaperones Human genes 0.000 claims description 38
- 238000013518 transcription Methods 0.000 claims description 36
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 35
- 230000035897 transcription Effects 0.000 claims description 35
- 210000004899 c-terminal region Anatomy 0.000 claims description 30
- 230000002068 genetic effect Effects 0.000 claims description 29
- 230000001580 bacterial effect Effects 0.000 claims description 25
- 239000003102 growth factor Substances 0.000 claims description 23
- 230000014759 maintenance of location Effects 0.000 claims description 21
- 229960004441 tyrosine Drugs 0.000 claims description 17
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 claims description 16
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 claims description 16
- 239000013613 expression plasmid Substances 0.000 claims description 15
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 claims description 14
- 229960001153 serine Drugs 0.000 claims description 10
- 108010002350 Interleukin-2 Proteins 0.000 claims description 9
- 102000000588 Interleukin-2 Human genes 0.000 claims description 9
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 9
- 230000002757 inflammatory effect Effects 0.000 claims description 8
- 229960005190 phenylalanine Drugs 0.000 claims description 8
- 102000004127 Cytokines Human genes 0.000 claims description 7
- 108090000695 Cytokines Proteins 0.000 claims description 7
- 102000004877 Insulin Human genes 0.000 claims description 7
- 108090001061 Insulin Proteins 0.000 claims description 7
- 102000003814 Interleukin-10 Human genes 0.000 claims description 7
- 108090000174 Interleukin-10 Proteins 0.000 claims description 7
- 102000013462 Interleukin-12 Human genes 0.000 claims description 7
- 108010065805 Interleukin-12 Proteins 0.000 claims description 7
- 108010002386 Interleukin-3 Proteins 0.000 claims description 7
- 102000015696 Interleukins Human genes 0.000 claims description 7
- 108010063738 Interleukins Proteins 0.000 claims description 7
- 229940125396 insulin Drugs 0.000 claims description 7
- 102000005755 Intercellular Signaling Peptides and Proteins Human genes 0.000 claims description 6
- 108010070716 Intercellular Signaling Peptides and Proteins Proteins 0.000 claims description 6
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 6
- 102000016267 Leptin Human genes 0.000 claims description 6
- 108010092277 Leptin Proteins 0.000 claims description 6
- 102000048850 Neoplasm Genes Human genes 0.000 claims description 6
- 108700019961 Neoplasm Genes Proteins 0.000 claims description 6
- NRYBAZVQPHGZNS-ZSOCWYAHSA-N leptin Chemical compound O=C([C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CC(C)C)CCSC)N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CS)C(O)=O NRYBAZVQPHGZNS-ZSOCWYAHSA-N 0.000 claims description 6
- 229940039781 leptin Drugs 0.000 claims description 6
- 239000000813 peptide hormone Substances 0.000 claims description 6
- BZQFBWGGLXLEPQ-REOHCLBHSA-N phosphoserine Chemical compound OC(=O)[C@@H](N)COP(O)(O)=O BZQFBWGGLXLEPQ-REOHCLBHSA-N 0.000 claims description 6
- CMUHFUGDYMFHEI-QMMMGPOBSA-N 4-amino-L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(N)C=C1 CMUHFUGDYMFHEI-QMMMGPOBSA-N 0.000 claims description 5
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 claims description 5
- 108050005493 CD3 protein, epsilon/gamma/delta subunit Proteins 0.000 claims description 5
- 108090000386 Fibroblast Growth Factor 1 Proteins 0.000 claims description 5
- 102000003971 Fibroblast Growth Factor 1 Human genes 0.000 claims description 5
- 102100031734 Fibroblast growth factor 19 Human genes 0.000 claims description 5
- 108090000376 Fibroblast growth factor 21 Proteins 0.000 claims description 5
- 102000003973 Fibroblast growth factor 21 Human genes 0.000 claims description 5
- 101800001586 Ghrelin Proteins 0.000 claims description 5
- 102400000442 Ghrelin-28 Human genes 0.000 claims description 5
- 102100041003 Glutamate carboxypeptidase 2 Human genes 0.000 claims description 5
- 102100032530 Glypican-3 Human genes 0.000 claims description 5
- 101000846394 Homo sapiens Fibroblast growth factor 19 Proteins 0.000 claims description 5
- 101000892862 Homo sapiens Glutamate carboxypeptidase 2 Proteins 0.000 claims description 5
- 101001014668 Homo sapiens Glypican-3 Proteins 0.000 claims description 5
- 101001103039 Homo sapiens Inactive tyrosine-protein kinase transmembrane receptor ROR1 Proteins 0.000 claims description 5
- 101000599951 Homo sapiens Insulin-like growth factor I Proteins 0.000 claims description 5
- 101001103036 Homo sapiens Nuclear receptor ROR-alpha Proteins 0.000 claims description 5
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 claims description 5
- 101100369992 Homo sapiens TNFSF10 gene Proteins 0.000 claims description 5
- 101000801433 Homo sapiens Trophoblast glycoprotein Proteins 0.000 claims description 5
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 claims description 5
- 102100037852 Insulin-like growth factor I Human genes 0.000 claims description 5
- 102000003812 Interleukin-15 Human genes 0.000 claims description 5
- 108090000172 Interleukin-15 Proteins 0.000 claims description 5
- 102000000646 Interleukin-3 Human genes 0.000 claims description 5
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 claims description 5
- 210000001744 T-lymphocyte Anatomy 0.000 claims description 5
- 102000046283 TNF-Related Apoptosis-Inducing Ligand Human genes 0.000 claims description 5
- 108700012411 TNFSF10 Proteins 0.000 claims description 5
- 101150117918 Tacstd2 gene Proteins 0.000 claims description 5
- 101710187743 Tumor necrosis factor receptor superfamily member 1A Proteins 0.000 claims description 5
- 102100033732 Tumor necrosis factor receptor superfamily member 1A Human genes 0.000 claims description 5
- 102100027212 Tumor-associated calcium signal transducer 2 Human genes 0.000 claims description 5
- 102000052116 epidermal growth factor receptor activity proteins Human genes 0.000 claims description 5
- 108700015053 epidermal growth factor receptor activity proteins Proteins 0.000 claims description 5
- GNKDKYIHGQKHHM-RJKLHVOGSA-N ghrelin Chemical compound C([C@H](NC(=O)[C@@H](NC(=O)[C@H](CO)NC(=O)CN)COC(=O)CCCCCCC)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C1=CC=CC=C1 GNKDKYIHGQKHHM-RJKLHVOGSA-N 0.000 claims description 5
- 108010074108 interleukin-21 Proteins 0.000 claims description 5
- YOHYSYJDKVYCJI-UHFFFAOYSA-N n-[3-[[6-[3-(trifluoromethyl)anilino]pyrimidin-4-yl]amino]phenyl]cyclopropanecarboxamide Chemical compound FC(F)(F)C1=CC=CC(NC=2N=CN=C(NC=3C=C(NC(=O)C4CC4)C=CC=3)C=2)=C1 YOHYSYJDKVYCJI-UHFFFAOYSA-N 0.000 claims description 5
- 229920001481 poly(stearyl methacrylate) Polymers 0.000 claims description 5
- YYTDJPUFAVPHQA-VKHMYHEASA-N (2s)-2-amino-3-(2,3,4,5,6-pentafluorophenyl)propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=C(F)C(F)=C(F)C(F)=C1F YYTDJPUFAVPHQA-VKHMYHEASA-N 0.000 claims description 4
- PEMUHKUIQHFMTH-QMMMGPOBSA-N (2s)-2-amino-3-(4-bromophenyl)propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=C(Br)C=C1 PEMUHKUIQHFMTH-QMMMGPOBSA-N 0.000 claims description 4
- NEMHIKRLROONTL-QMMMGPOBSA-N (2s)-2-azaniumyl-3-(4-azidophenyl)propanoate Chemical compound OC(=O)[C@@H](N)CC1=CC=C(N=[N+]=[N-])C=C1 NEMHIKRLROONTL-QMMMGPOBSA-N 0.000 claims description 4
- JZRBSTONIYRNRI-VIFPVBQESA-N 3-methylphenylalanine Chemical compound CC1=CC=CC(C[C@H](N)C(O)=O)=C1 JZRBSTONIYRNRI-VIFPVBQESA-N 0.000 claims description 4
- IRZQDMYEJPNDEN-UHFFFAOYSA-N 3-phenyl-2-aminobutanoic acid Natural products OC(=O)C(N)C(C)C1=CC=CC=C1 IRZQDMYEJPNDEN-UHFFFAOYSA-N 0.000 claims description 4
- PZNQZSRPDOEBMS-QMMMGPOBSA-N 4-iodo-L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(I)C=C1 PZNQZSRPDOEBMS-QMMMGPOBSA-N 0.000 claims description 4
- 102100036466 Delta-like protein 3 Human genes 0.000 claims description 4
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 claims description 4
- 108090000569 Fibroblast Growth Factor-23 Proteins 0.000 claims description 4
- 102100024802 Fibroblast growth factor 23 Human genes 0.000 claims description 4
- 101000928513 Homo sapiens Delta-like protein 3 Proteins 0.000 claims description 4
- 102000017727 Immunoglobulin Variable Region Human genes 0.000 claims description 4
- WTDRDQBEARUVNC-LURJTMIESA-N L-DOPA Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-LURJTMIESA-N 0.000 claims description 4
- WTDRDQBEARUVNC-UHFFFAOYSA-N L-Dopa Natural products OC(=O)C(N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-UHFFFAOYSA-N 0.000 claims description 4
- GEYBMYRBIABFTA-VIFPVBQESA-N O-methyl-L-tyrosine Chemical compound COC1=CC=C(C[C@H](N)C(O)=O)C=C1 GEYBMYRBIABFTA-VIFPVBQESA-N 0.000 claims description 4
- 102100033579 Trophoblast glycoprotein Human genes 0.000 claims description 4
- 229940127276 delta-like ligand 3 Drugs 0.000 claims description 4
- 229950006137 dexfosfoserine Drugs 0.000 claims description 4
- 238000010839 reverse transcription Methods 0.000 claims description 4
- 230000011664 signaling Effects 0.000 claims description 4
- 102000005969 steroid hormone receptors Human genes 0.000 claims description 4
- 108020003113 steroid hormone receptors Proteins 0.000 claims description 4
- JPZXHKDZASGCLU-LBPRGKRZSA-N β-(2-naphthyl)-alanine Chemical compound C1=CC=CC2=CC(C[C@H](N)C(O)=O)=CC=C21 JPZXHKDZASGCLU-LBPRGKRZSA-N 0.000 claims description 4
- 102000001617 Interferon Receptors Human genes 0.000 claims description 3
- 108010054267 Interferon Receptors Proteins 0.000 claims description 3
- 102100030703 Interleukin-22 Human genes 0.000 claims description 3
- TVIDEEHSOPHZBR-AWEZNQCLSA-N para-(benzoyl)-phenylalanine Chemical compound C1=CC(C[C@H](N)C(O)=O)=CC=C1C(=O)C1=CC=CC=C1 TVIDEEHSOPHZBR-AWEZNQCLSA-N 0.000 claims description 3
- 238000000638 solvent extraction Methods 0.000 claims description 3
- 229960005486 vaccine Drugs 0.000 claims description 3
- POGSZHUEECCEAP-ZETCQYMHSA-N (2s)-2-amino-3-(3-amino-4-hydroxyphenyl)propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C(N)=C1 POGSZHUEECCEAP-ZETCQYMHSA-N 0.000 claims description 2
- 102000019034 Chemokines Human genes 0.000 claims description 2
- 108010012236 Chemokines Proteins 0.000 claims description 2
- OUGDEOOFDGWUAC-QMMMGPOBSA-N (2S)-2-amino-3-(3-sulfooxyphenyl)propanoic acid Chemical compound S(=O)(=O)(O)OC=1C=C(C[C@H](N)C(=O)O)C=CC1 OUGDEOOFDGWUAC-QMMMGPOBSA-N 0.000 claims 1
- DJZAVKNBQZFEAL-QMMMGPOBSA-N (2s)-2-amino-3-(2-aminophenyl)propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1N DJZAVKNBQZFEAL-QMMMGPOBSA-N 0.000 claims 1
- QTCRLZGTYUJQIE-ZETCQYMHSA-N (2s)-2-amino-3-(2-sulfooxyphenyl)propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1OS(O)(=O)=O QTCRLZGTYUJQIE-ZETCQYMHSA-N 0.000 claims 1
- YDYSBABSIVBERF-ZETCQYMHSA-N (2s)-2-amino-3-(2-sulfophenyl)propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1S(O)(=O)=O YDYSBABSIVBERF-ZETCQYMHSA-N 0.000 claims 1
- DDLYNVBJVVOUGB-QMMMGPOBSA-N (2s)-2-amino-3-(3-aminophenyl)propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=CC(N)=C1 DDLYNVBJVVOUGB-QMMMGPOBSA-N 0.000 claims 1
- SUPCVWGCSKIXFE-QMMMGPOBSA-N (2s)-2-amino-3-(3-sulfophenyl)propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=CC(S(O)(=O)=O)=C1 SUPCVWGCSKIXFE-QMMMGPOBSA-N 0.000 claims 1
- BJOQKIKXKGJLIJ-NSHDSACASA-N (2s)-2-amino-3-(4-prop-2-ynylphenyl)propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=C(CC#C)C=C1 BJOQKIKXKGJLIJ-NSHDSACASA-N 0.000 claims 1
- ALQIUGWFHKQQHV-QMMMGPOBSA-N (2s)-2-amino-3-(4-sulfophenyl)propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=C(S(O)(=O)=O)C=C1 ALQIUGWFHKQQHV-QMMMGPOBSA-N 0.000 claims 1
- SDZGVFSSLGTJAJ-ZETCQYMHSA-N (2s)-2-azaniumyl-3-(2-nitrophenyl)propanoate Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1[N+]([O-])=O SDZGVFSSLGTJAJ-ZETCQYMHSA-N 0.000 claims 1
- YTHDRUZHNYKZGF-QMMMGPOBSA-N (2s)-2-azaniumyl-3-(3-nitrophenyl)propanoate Chemical compound OC(=O)[C@@H](N)CC1=CC=CC([N+]([O-])=O)=C1 YTHDRUZHNYKZGF-QMMMGPOBSA-N 0.000 claims 1
- PQUSMXUQSMSWNN-QMMMGPOBSA-N (2s)-3-(2-amino-4-hydroxyphenyl)-2-azaniumylpropanoate Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1N PQUSMXUQSMSWNN-QMMMGPOBSA-N 0.000 claims 1
- ZXSBHXZKWRIEIA-JTQLQIEISA-N (2s)-3-(4-acetylphenyl)-2-azaniumylpropanoate Chemical compound CC(=O)C1=CC=C(C[C@H](N)C(O)=O)C=C1 ZXSBHXZKWRIEIA-JTQLQIEISA-N 0.000 claims 1
- DXAUAWUGCKCSFC-NSHDSACASA-N (2s)-3-phenyl-2-(prop-2-ynoxyamino)propanoic acid Chemical compound C#CCON[C@H](C(=O)O)CC1=CC=CC=C1 DXAUAWUGCKCSFC-NSHDSACASA-N 0.000 claims 1
- SXIFEQOVCDBNET-QMMMGPOBSA-N 2-[(2s)-2-amino-2-carboxyethyl]benzoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1C(O)=O SXIFEQOVCDBNET-QMMMGPOBSA-N 0.000 claims 1
- QDGAVODICPCDMU-UHFFFAOYSA-N 2-amino-3-[3-[bis(2-chloroethyl)amino]phenyl]propanoic acid Chemical compound OC(=O)C(N)CC1=CC=CC(N(CCCl)CCCl)=C1 QDGAVODICPCDMU-UHFFFAOYSA-N 0.000 claims 1
- JANONUPBHGWOBP-QMMMGPOBSA-N 3-[(2s)-2-amino-2-carboxyethyl]benzoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=CC(C(O)=O)=C1 JANONUPBHGWOBP-QMMMGPOBSA-N 0.000 claims 1
- YXDGRBPZVQPESQ-QMMMGPOBSA-N 4-[(2s)-2-amino-2-carboxyethyl]benzoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=C(C(O)=O)C=C1 YXDGRBPZVQPESQ-QMMMGPOBSA-N 0.000 claims 1
- GTVVZTAFGPQSPC-UHFFFAOYSA-N 4-nitrophenylalanine Chemical compound OC(=O)C(N)CC1=CC=C([N+]([O-])=O)C=C1 GTVVZTAFGPQSPC-UHFFFAOYSA-N 0.000 claims 1
- 108010092408 Eosinophil Peroxidase Proteins 0.000 claims 1
- 102100028471 Eosinophil peroxidase Human genes 0.000 claims 1
- 102100039614 Nuclear receptor ROR-alpha Human genes 0.000 claims 1
- CIQHWLTYGMYQQR-QMMMGPOBSA-N O(4')-sulfo-L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(OS(O)(=O)=O)C=C1 CIQHWLTYGMYQQR-QMMMGPOBSA-N 0.000 claims 1
- BZQFBWGGLXLEPQ-UHFFFAOYSA-N O-phosphoryl-L-serine Natural products OC(=O)C(N)COP(O)(O)=O BZQFBWGGLXLEPQ-UHFFFAOYSA-N 0.000 claims 1
- 238000005192 partition Methods 0.000 claims 1
- 239000000203 mixture Substances 0.000 abstract description 27
- 230000001965 increasing effect Effects 0.000 abstract description 15
- 239000004615 ingredient Substances 0.000 abstract description 6
- 108020004705 Codon Proteins 0.000 description 84
- 241000588724 Escherichia coli Species 0.000 description 66
- 102000039446 nucleic acids Human genes 0.000 description 64
- 108020004707 nucleic acids Proteins 0.000 description 64
- 238000004519 manufacturing process Methods 0.000 description 55
- 108020004414 DNA Proteins 0.000 description 43
- 238000000855 fermentation Methods 0.000 description 38
- 230000004151 fermentation Effects 0.000 description 38
- 108020004566 Transfer RNA Proteins 0.000 description 27
- 230000014616 translation Effects 0.000 description 27
- 241000894006 Bacteria Species 0.000 description 26
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 26
- 102000040430 polynucleotide Human genes 0.000 description 25
- 108091033319 polynucleotide Proteins 0.000 description 25
- 239000002157 polynucleotide Substances 0.000 description 25
- 238000013519 translation Methods 0.000 description 24
- 108091008146 restriction endonucleases Proteins 0.000 description 23
- 230000000694 effects Effects 0.000 description 22
- 239000008188 pellet Substances 0.000 description 21
- 239000002609 medium Substances 0.000 description 20
- 230000015572 biosynthetic process Effects 0.000 description 19
- 238000001727 in vivo Methods 0.000 description 19
- 230000004048 modification Effects 0.000 description 19
- 238000012986 modification Methods 0.000 description 19
- 239000000047 product Substances 0.000 description 17
- 239000000126 substance Chemical class 0.000 description 16
- 125000003275 alpha amino acid group Chemical group 0.000 description 15
- 230000012010 growth Effects 0.000 description 15
- 238000000338 in vitro Methods 0.000 description 15
- 239000003550 marker Substances 0.000 description 15
- 239000002773 nucleotide Substances 0.000 description 15
- 125000003729 nucleotide group Chemical group 0.000 description 15
- 230000008569 process Effects 0.000 description 15
- 239000000523 sample Substances 0.000 description 15
- 108010076504 Protein Sorting Signals Proteins 0.000 description 14
- 238000010790 dilution Methods 0.000 description 14
- 239000012895 dilution Substances 0.000 description 14
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 13
- 238000012217 deletion Methods 0.000 description 13
- 230000037430 deletion Effects 0.000 description 13
- 210000001322 periplasm Anatomy 0.000 description 13
- 239000013604 expression vector Substances 0.000 description 12
- 230000017854 proteolysis Effects 0.000 description 12
- 230000001105 regulatory effect Effects 0.000 description 12
- 238000003786 synthesis reaction Methods 0.000 description 12
- 108091026890 Coding region Proteins 0.000 description 11
- 238000010367 cloning Methods 0.000 description 11
- 238000005516 engineering process Methods 0.000 description 11
- 230000012846 protein folding Effects 0.000 description 11
- 230000002829 reductive effect Effects 0.000 description 11
- 241000894007 species Species 0.000 description 11
- 102000018233 Fibroblast Growth Factor Human genes 0.000 description 10
- 108050007372 Fibroblast Growth Factor Proteins 0.000 description 10
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 10
- 102100020880 Kit ligand Human genes 0.000 description 10
- 102000035195 Peptidases Human genes 0.000 description 10
- 108091005804 Peptidases Proteins 0.000 description 10
- 239000004365 Protease Substances 0.000 description 10
- 102000013275 Somatomedins Human genes 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 10
- 238000003776 cleavage reaction Methods 0.000 description 10
- 230000004186 co-expression Effects 0.000 description 10
- 150000001875 compounds Chemical class 0.000 description 10
- 238000013461 design Methods 0.000 description 10
- 229940126864 fibroblast growth factor Drugs 0.000 description 10
- 239000008103 glucose Substances 0.000 description 10
- 229930027917 kanamycin Natural products 0.000 description 10
- 229960000318 kanamycin Drugs 0.000 description 10
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 10
- 229930182823 kanamycin A Natural products 0.000 description 10
- 239000000463 material Substances 0.000 description 10
- 230000002441 reversible effect Effects 0.000 description 10
- 230000007017 scission Effects 0.000 description 10
- 238000011144 upstream manufacturing Methods 0.000 description 10
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 9
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 9
- 230000033228 biological regulation Effects 0.000 description 9
- 239000000872 buffer Substances 0.000 description 9
- 239000003153 chemical reaction reagent Substances 0.000 description 9
- 239000003795 chemical substances by application Substances 0.000 description 9
- 230000001939 inductive effect Effects 0.000 description 9
- 230000004481 post-translational protein modification Effects 0.000 description 9
- 238000000746 purification Methods 0.000 description 9
- 238000006467 substitution reaction Methods 0.000 description 9
- 230000001629 suppression Effects 0.000 description 9
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 8
- 102000053602 DNA Human genes 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 8
- 108090000790 Enzymes Proteins 0.000 description 8
- 108090000385 Fibroblast growth factor 7 Proteins 0.000 description 8
- 102000003972 Fibroblast growth factor 7 Human genes 0.000 description 8
- 108020005038 Terminator Codon Proteins 0.000 description 8
- 239000004098 Tetracycline Substances 0.000 description 8
- 229940088598 enzyme Drugs 0.000 description 8
- 239000003112 inhibitor Substances 0.000 description 8
- 210000000440 neutrophil Anatomy 0.000 description 8
- 229920000642 polymer Polymers 0.000 description 8
- 102000005962 receptors Human genes 0.000 description 8
- 108020003175 receptors Proteins 0.000 description 8
- 230000010076 replication Effects 0.000 description 8
- 238000004007 reversed phase HPLC Methods 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 229960002180 tetracycline Drugs 0.000 description 8
- 229930101283 tetracycline Natural products 0.000 description 8
- 235000019364 tetracycline Nutrition 0.000 description 8
- 150000003522 tetracyclines Chemical class 0.000 description 8
- 102000052866 Amino Acyl-tRNA Synthetases Human genes 0.000 description 7
- 108700028939 Amino Acyl-tRNA Synthetases Proteins 0.000 description 7
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 7
- 238000006664 bond formation reaction Methods 0.000 description 7
- 210000000805 cytoplasm Anatomy 0.000 description 7
- 230000001976 improved effect Effects 0.000 description 7
- 230000010354 integration Effects 0.000 description 7
- 238000006317 isomerization reaction Methods 0.000 description 7
- OOYGSFOGFJDDHP-KMCOLRRFSA-N kanamycin A sulfate Chemical compound OS(O)(=O)=O.O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N OOYGSFOGFJDDHP-KMCOLRRFSA-N 0.000 description 7
- 230000002147 killing effect Effects 0.000 description 7
- 108020004999 messenger RNA Proteins 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 108090000626 DNA-directed RNA polymerases Proteins 0.000 description 6
- 238000002965 ELISA Methods 0.000 description 6
- 241000206602 Eukaryota Species 0.000 description 6
- OAKJQQAXSVQMHS-UHFFFAOYSA-N Hydrazine Chemical compound NN OAKJQQAXSVQMHS-UHFFFAOYSA-N 0.000 description 6
- 102100025390 Integrin beta-2 Human genes 0.000 description 6
- 108010064593 Intercellular Adhesion Molecule-1 Proteins 0.000 description 6
- 102100037877 Intercellular adhesion molecule 1 Human genes 0.000 description 6
- 101710177504 Kit ligand Proteins 0.000 description 6
- 108010064548 Lymphocyte Function-Associated Antigen-1 Proteins 0.000 description 6
- 108091005461 Nucleic proteins Proteins 0.000 description 6
- 101100084022 Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) lapA gene Proteins 0.000 description 6
- RWRDLPDLKQPQOW-UHFFFAOYSA-N Pyrrolidine Chemical compound C1CCNC1 RWRDLPDLKQPQOW-UHFFFAOYSA-N 0.000 description 6
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 6
- 108060008683 Tumor Necrosis Factor Receptor Proteins 0.000 description 6
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 6
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 6
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 6
- 230000003213 activating effect Effects 0.000 description 6
- 125000000539 amino acid group Chemical group 0.000 description 6
- FPPNZSSZRUTDAP-UWFZAAFLSA-N carbenicillin Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)C(C(O)=O)C1=CC=CC=C1 FPPNZSSZRUTDAP-UWFZAAFLSA-N 0.000 description 6
- 229960003669 carbenicillin Drugs 0.000 description 6
- 238000005277 cation exchange chromatography Methods 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 238000003306 harvesting Methods 0.000 description 6
- 238000010348 incorporation Methods 0.000 description 6
- 230000006698 induction Effects 0.000 description 6
- 238000003780 insertion Methods 0.000 description 6
- 230000037431 insertion Effects 0.000 description 6
- 229960002064 kanamycin sulfate Drugs 0.000 description 6
- 238000004949 mass spectrometry Methods 0.000 description 6
- 230000035772 mutation Effects 0.000 description 6
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 6
- 101150009573 phoA gene Proteins 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 101150003695 proS gene Proteins 0.000 description 6
- 230000009467 reduction Effects 0.000 description 6
- 230000002103 transcriptional effect Effects 0.000 description 6
- 102000003298 tumor necrosis factor receptor Human genes 0.000 description 6
- 108020005098 Anticodon Proteins 0.000 description 5
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 5
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 5
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 5
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 5
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 5
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 5
- 102000009658 Peptidylprolyl Isomerase Human genes 0.000 description 5
- 108010020062 Peptidylprolyl Isomerase Proteins 0.000 description 5
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 5
- 102000006010 Protein Disulfide-Isomerase Human genes 0.000 description 5
- 108020004511 Recombinant DNA Proteins 0.000 description 5
- 239000002253 acid Substances 0.000 description 5
- 235000004279 alanine Nutrition 0.000 description 5
- 239000003242 anti bacterial agent Substances 0.000 description 5
- 229940088710 antibiotic agent Drugs 0.000 description 5
- 238000013459 approach Methods 0.000 description 5
- 238000003556 assay Methods 0.000 description 5
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 5
- 230000000295 complement effect Effects 0.000 description 5
- 238000010276 construction Methods 0.000 description 5
- 239000003398 denaturant Substances 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 238000012063 dual-affinity re-targeting Methods 0.000 description 5
- 238000010828 elution Methods 0.000 description 5
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 5
- 235000004554 glutamine Nutrition 0.000 description 5
- 208000032839 leukemia Diseases 0.000 description 5
- 229930182817 methionine Natural products 0.000 description 5
- 239000001301 oxygen Substances 0.000 description 5
- 229910052760 oxygen Inorganic materials 0.000 description 5
- 230000027086 plasmid maintenance Effects 0.000 description 5
- 235000013930 proline Nutrition 0.000 description 5
- 108020003519 protein disulfide isomerase Proteins 0.000 description 5
- 230000002285 radioactive effect Effects 0.000 description 5
- 230000028327 secretion Effects 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 239000002904 solvent Substances 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 239000004094 surface-active agent Substances 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- 238000012546 transfer Methods 0.000 description 5
- 230000009466 transformation Effects 0.000 description 5
- 150000003668 tyrosines Chemical class 0.000 description 5
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 4
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 4
- BNIFSVVAHBLNTN-XKKUQSFHSA-N (2s)-4-amino-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-1-[(2s)-4-amino-2-[[2-[[(2s)-2-[[(2s)-2-[[(2s)-1-[(2s)-6-amino-2-[[(2s)-2-[[(2s)-2-[[(2s,3r)-2-amino-3-hydroxybutanoyl]amino]-4-methylsulfanylbutanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]hexan Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(=O)N1[C@H](C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O)CCC1 BNIFSVVAHBLNTN-XKKUQSFHSA-N 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 4
- 239000004475 Arginine Substances 0.000 description 4
- FDKWRPBBCBCIGA-UWTATZPHSA-N D-Selenocysteine Natural products [Se]C[C@@H](N)C(O)=O FDKWRPBBCBCIGA-UWTATZPHSA-N 0.000 description 4
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 description 4
- 102000004269 Granulocyte Colony-Stimulating Factor Human genes 0.000 description 4
- 102100034221 Growth-regulated alpha protein Human genes 0.000 description 4
- 102000002265 Human Growth Hormone Human genes 0.000 description 4
- 108010000521 Human Growth Hormone Proteins 0.000 description 4
- 239000000854 Human Growth Hormone Substances 0.000 description 4
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 4
- 102100039615 Inactive tyrosine-protein kinase transmembrane receptor ROR1 Human genes 0.000 description 4
- 102000048143 Insulin-Like Growth Factor II Human genes 0.000 description 4
- 108090001117 Insulin-Like Growth Factor II Proteins 0.000 description 4
- 108010008212 Integrin alpha4beta1 Proteins 0.000 description 4
- 102000014150 Interferons Human genes 0.000 description 4
- 108010050904 Interferons Proteins 0.000 description 4
- 102000000589 Interleukin-1 Human genes 0.000 description 4
- 108010002352 Interleukin-1 Proteins 0.000 description 4
- 108090001007 Interleukin-8 Proteins 0.000 description 4
- 102000004890 Interleukin-8 Human genes 0.000 description 4
- 229930194542 Keto Chemical class 0.000 description 4
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 4
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 4
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 4
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 4
- ZKZBPNGNEQAJSX-REOHCLBHSA-N L-selenocysteine Chemical compound [SeH]C[C@H](N)C(O)=O ZKZBPNGNEQAJSX-REOHCLBHSA-N 0.000 description 4
- 108010001831 LDL receptors Proteins 0.000 description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 4
- 239000004472 Lysine Substances 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 4
- 108090000375 Mineralocorticoid Receptors Proteins 0.000 description 4
- 102000003979 Mineralocorticoid Receptors Human genes 0.000 description 4
- 108020004485 Nonsense Codon Proteins 0.000 description 4
- 108091060545 Nonsense suppressor Proteins 0.000 description 4
- 239000004721 Polyphenylene oxide Substances 0.000 description 4
- 241000589540 Pseudomonas fluorescens Species 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 108010039445 Stem Cell Factor Proteins 0.000 description 4
- 101710137500 T7 RNA polymerase Proteins 0.000 description 4
- 239000004473 Threonine Substances 0.000 description 4
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 4
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 4
- 108010009583 Transforming Growth Factors Proteins 0.000 description 4
- 102000009618 Transforming Growth Factors Human genes 0.000 description 4
- 101800004564 Transforming growth factor alpha Proteins 0.000 description 4
- 102400001320 Transforming growth factor alpha Human genes 0.000 description 4
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 4
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 4
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 108010000134 Vascular Cell Adhesion Molecule-1 Proteins 0.000 description 4
- 102100023543 Vascular cell adhesion protein 1 Human genes 0.000 description 4
- 241000700605 Viruses Species 0.000 description 4
- 239000008272 agar Substances 0.000 description 4
- 108010080146 androgen receptors Proteins 0.000 description 4
- 102000001307 androgen receptors Human genes 0.000 description 4
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 4
- 235000009697 arginine Nutrition 0.000 description 4
- 230000001746 atrial effect Effects 0.000 description 4
- 230000008901 benefit Effects 0.000 description 4
- 230000001588 bifunctional effect Effects 0.000 description 4
- 230000003197 catalytic effect Effects 0.000 description 4
- 230000032823 cell division Effects 0.000 description 4
- 230000001413 cellular effect Effects 0.000 description 4
- 210000000349 chromosome Anatomy 0.000 description 4
- 230000009089 cytolysis Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 230000008472 epithelial growth Effects 0.000 description 4
- 210000003527 eukaryotic cell Anatomy 0.000 description 4
- 101150027203 fkpA gene Proteins 0.000 description 4
- 230000037433 frameshift Effects 0.000 description 4
- 150000002308 glutamine derivatives Chemical class 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 210000003000 inclusion body Anatomy 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 125000000468 ketone group Chemical class 0.000 description 4
- 229910052751 metal Inorganic materials 0.000 description 4
- 239000002184 metal Substances 0.000 description 4
- 230000000813 microbial effect Effects 0.000 description 4
- 238000010369 molecular cloning Methods 0.000 description 4
- 210000004940 nucleus Anatomy 0.000 description 4
- 238000005457 optimization Methods 0.000 description 4
- 230000003647 oxidation Effects 0.000 description 4
- 238000007254 oxidation reaction Methods 0.000 description 4
- 230000037361 pathway Effects 0.000 description 4
- 108010012038 peptide 78 Proteins 0.000 description 4
- 229940125863 peptide 78 Drugs 0.000 description 4
- 150000003904 phospholipids Chemical class 0.000 description 4
- 229920000570 polyether Polymers 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 238000012545 processing Methods 0.000 description 4
- 210000001236 prokaryotic cell Anatomy 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- 229940055619 selenocysteine Drugs 0.000 description 4
- ZKZBPNGNEQAJSX-UHFFFAOYSA-N selenocysteine Natural products [SeH]CC(N)C(O)=O ZKZBPNGNEQAJSX-UHFFFAOYSA-N 0.000 description 4
- 235000016491 selenocysteine Nutrition 0.000 description 4
- 229910052708 sodium Inorganic materials 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 238000010561 standard procedure Methods 0.000 description 4
- 235000000346 sugar Nutrition 0.000 description 4
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 4
- 229960002898 threonine Drugs 0.000 description 4
- 239000003053 toxin Substances 0.000 description 4
- 231100000765 toxin Toxicity 0.000 description 4
- 108700012359 toxins Proteins 0.000 description 4
- 238000010361 transduction Methods 0.000 description 4
- 230000026683 transduction Effects 0.000 description 4
- 108020005087 unfolded proteins Proteins 0.000 description 4
- 241001515965 unidentified phage Species 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical class [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 3
- 108010071942 Colony-Stimulating Factors Proteins 0.000 description 3
- OMFXVFTZEKFJBZ-UHFFFAOYSA-N Corticosterone Natural products O=C1CCC2(C)C3C(O)CC(C)(C(CC4)C(=O)CO)C4C3CCC2=C1 OMFXVFTZEKFJBZ-UHFFFAOYSA-N 0.000 description 3
- 238000001712 DNA sequencing Methods 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 3
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 3
- 108010051696 Growth Hormone Proteins 0.000 description 3
- 102000018997 Growth Hormone Human genes 0.000 description 3
- 241000238631 Hexapoda Species 0.000 description 3
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical class ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 description 3
- 101000668058 Infectious salmon anemia virus (isolate Atlantic salmon/Norway/810/9/99) RNA-directed RNA polymerase catalytic subunit Proteins 0.000 description 3
- 102000004218 Insulin-Like Growth Factor I Human genes 0.000 description 3
- 102000004195 Isomerases Human genes 0.000 description 3
- 108090000769 Isomerases Proteins 0.000 description 3
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 3
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 3
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 3
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 3
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 3
- 102100024640 Low-density lipoprotein receptor Human genes 0.000 description 3
- 108091034117 Oligonucleotide Proteins 0.000 description 3
- 229910019142 PO4 Inorganic materials 0.000 description 3
- 101150056612 PPIA gene Proteins 0.000 description 3
- 101100533690 Pasteurella multocida (strain Pm70) slyD gene Proteins 0.000 description 3
- 241000589516 Pseudomonas Species 0.000 description 3
- 241000589776 Pseudomonas putida Species 0.000 description 3
- 108090000783 Renin Proteins 0.000 description 3
- 102100028255 Renin Human genes 0.000 description 3
- 102000019197 Superoxide Dismutase Human genes 0.000 description 3
- 108010012715 Superoxide dismutase Proteins 0.000 description 3
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 3
- 102000003978 Tissue Plasminogen Activator Human genes 0.000 description 3
- 108090000373 Tissue Plasminogen Activator Proteins 0.000 description 3
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 3
- 108091006088 activator proteins Proteins 0.000 description 3
- 230000002776 aggregation Effects 0.000 description 3
- 238000004220 aggregation Methods 0.000 description 3
- 239000000427 antigen Substances 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 235000003704 aspartic acid Nutrition 0.000 description 3
- 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 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 239000013592 cell lysate Substances 0.000 description 3
- 239000003638 chemical reducing agent Substances 0.000 description 3
- OMFXVFTZEKFJBZ-HJTSIMOOSA-N corticosterone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@H](CC4)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 OMFXVFTZEKFJBZ-HJTSIMOOSA-N 0.000 description 3
- 125000004122 cyclic group Chemical group 0.000 description 3
- 235000018417 cysteine Nutrition 0.000 description 3
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 3
- 230000001461 cytolytic effect Effects 0.000 description 3
- 238000004520 electroporation Methods 0.000 description 3
- 150000002148 esters Chemical class 0.000 description 3
- 102000015694 estrogen receptors Human genes 0.000 description 3
- 108010038795 estrogen receptors Proteins 0.000 description 3
- 239000012526 feed medium Substances 0.000 description 3
- 235000013922 glutamic acid Nutrition 0.000 description 3
- 239000004220 glutamic acid Substances 0.000 description 3
- 238000000265 homogenisation Methods 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 102000002467 interleukin receptors Human genes 0.000 description 3
- 108010093036 interleukin receptors Proteins 0.000 description 3
- 239000000543 intermediate Substances 0.000 description 3
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 3
- 230000000670 limiting effect Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 210000004962 mammalian cell Anatomy 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 3
- 238000013508 migration Methods 0.000 description 3
- 230000005012 migration Effects 0.000 description 3
- 229910052757 nitrogen Inorganic materials 0.000 description 3
- 239000007800 oxidant agent Substances 0.000 description 3
- 239000000546 pharmaceutical excipient Substances 0.000 description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 3
- 239000010452 phosphate Substances 0.000 description 3
- DCWXELXMIBXGTH-QMMMGPOBSA-N phosphonotyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(OP(O)(O)=O)C=C1 DCWXELXMIBXGTH-QMMMGPOBSA-N 0.000 description 3
- 230000004683 plasmid partitioning Effects 0.000 description 3
- 101150108030 ppiD gene Proteins 0.000 description 3
- 102000003998 progesterone receptors Human genes 0.000 description 3
- 108090000468 progesterone receptors Proteins 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 230000002797 proteolythic effect Effects 0.000 description 3
- 230000006798 recombination Effects 0.000 description 3
- 238000005215 recombination Methods 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 210000003705 ribosome Anatomy 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 238000002741 site-directed mutagenesis Methods 0.000 description 3
- 238000009331 sowing Methods 0.000 description 3
- 230000003595 spectral effect Effects 0.000 description 3
- 230000008685 targeting Effects 0.000 description 3
- 229960000187 tissue plasminogen activator Drugs 0.000 description 3
- 230000005026 transcription initiation Effects 0.000 description 3
- 125000001493 tyrosinyl group Chemical group [H]OC1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 3
- 239000013603 viral vector Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- YQAXFVHNHSPUPO-RNJOBUHISA-N 2-[[(2s)-2-[[2-[[(2s,4r)-1-[(2s)-1-(2-aminoacetyl)pyrrolidine-2-carbonyl]-4-hydroxypyrrolidine-2-carbonyl]amino]acetyl]amino]propanoyl]amino]acetic acid Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H]1C[C@@H](O)CN1C(=O)[C@H]1N(C(=O)CN)CCC1 YQAXFVHNHSPUPO-RNJOBUHISA-N 0.000 description 2
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 2
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 2
- 102400000068 Angiostatin Human genes 0.000 description 2
- 108010079709 Angiostatins Proteins 0.000 description 2
- 108010083590 Apoproteins Proteins 0.000 description 2
- 102000006410 Apoproteins Human genes 0.000 description 2
- 101000716807 Arabidopsis thaliana Protein SCO1 homolog 1, mitochondrial Proteins 0.000 description 2
- 241000203069 Archaea Species 0.000 description 2
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 2
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 description 2
- 102100032367 C-C motif chemokine 5 Human genes 0.000 description 2
- 102100025248 C-X-C motif chemokine 10 Human genes 0.000 description 2
- 101710098275 C-X-C motif chemokine 10 Proteins 0.000 description 2
- 102100039398 C-X-C motif chemokine 2 Human genes 0.000 description 2
- 102100036150 C-X-C motif chemokine 5 Human genes 0.000 description 2
- 102100036153 C-X-C motif chemokine 6 Human genes 0.000 description 2
- 101710085504 C-X-C motif chemokine 6 Proteins 0.000 description 2
- 102100036170 C-X-C motif chemokine 9 Human genes 0.000 description 2
- 101710085500 C-X-C motif chemokine 9 Proteins 0.000 description 2
- 108010078959 C-terminal processing peptidase Proteins 0.000 description 2
- 102000001902 CC Chemokines Human genes 0.000 description 2
- 108010040471 CC Chemokines Proteins 0.000 description 2
- 108700012434 CCL3 Proteins 0.000 description 2
- 108010029697 CD40 Ligand Proteins 0.000 description 2
- 101150013553 CD40 gene Proteins 0.000 description 2
- 102100032937 CD40 ligand Human genes 0.000 description 2
- 102100032912 CD44 antigen Human genes 0.000 description 2
- 101150093802 CXCL1 gene Proteins 0.000 description 2
- 102000055006 Calcitonin Human genes 0.000 description 2
- 108060001064 Calcitonin Proteins 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 102000000013 Chemokine CCL3 Human genes 0.000 description 2
- 108010055166 Chemokine CCL5 Proteins 0.000 description 2
- 108010014419 Chemokine CXCL1 Proteins 0.000 description 2
- 102000016950 Chemokine CXCL1 Human genes 0.000 description 2
- 102100022641 Coagulation factor IX Human genes 0.000 description 2
- 102100023804 Coagulation factor VII Human genes 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 2
- 108010035532 Collagen Proteins 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 101710104662 Enterotoxin type C-3 Proteins 0.000 description 2
- 102100030844 Exocyst complex component 1 Human genes 0.000 description 2
- 101710082714 Exotoxin A Proteins 0.000 description 2
- 108010076282 Factor IX Proteins 0.000 description 2
- 108010023321 Factor VII Proteins 0.000 description 2
- 108010014173 Factor X Proteins 0.000 description 2
- 108010049003 Fibrinogen Proteins 0.000 description 2
- 102000008946 Fibrinogen Human genes 0.000 description 2
- 108090000368 Fibroblast growth factor 8 Proteins 0.000 description 2
- 102100037362 Fibronectin Human genes 0.000 description 2
- 108010067306 Fibronectins Proteins 0.000 description 2
- 102100040837 Galactoside alpha-(1,2)-fucosyltransferase 2 Human genes 0.000 description 2
- 101710115997 Gamma-tubulin complex component 2 Proteins 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- 102000006771 Gonadotropins Human genes 0.000 description 2
- 108010086677 Gonadotropins Proteins 0.000 description 2
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 2
- ZRALSGWEFCBTJO-UHFFFAOYSA-N Guanidine Chemical compound NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 description 2
- 241000205062 Halobacterium Species 0.000 description 2
- 102000003693 Hedgehog Proteins Human genes 0.000 description 2
- 108090000031 Hedgehog Proteins Proteins 0.000 description 2
- 102000001554 Hemoglobins Human genes 0.000 description 2
- 108010054147 Hemoglobins Proteins 0.000 description 2
- 102000007625 Hirudins Human genes 0.000 description 2
- 108010007267 Hirudins Proteins 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 101000889128 Homo sapiens C-X-C motif chemokine 2 Proteins 0.000 description 2
- 101000947186 Homo sapiens C-X-C motif chemokine 5 Proteins 0.000 description 2
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 description 2
- 101000893710 Homo sapiens Galactoside alpha-(1,2)-fucosyltransferase 2 Proteins 0.000 description 2
- 101001069921 Homo sapiens Growth-regulated alpha protein Proteins 0.000 description 2
- 101000973997 Homo sapiens Nucleosome assembly protein 1-like 4 Proteins 0.000 description 2
- 101000947178 Homo sapiens Platelet basic protein Proteins 0.000 description 2
- 101000582950 Homo sapiens Platelet factor 4 Proteins 0.000 description 2
- 101001076715 Homo sapiens RNA-binding protein 39 Proteins 0.000 description 2
- 101000652229 Homo sapiens Suppressor of cytokine signaling 7 Proteins 0.000 description 2
- 101000611183 Homo sapiens Tumor necrosis factor Proteins 0.000 description 2
- 102000008100 Human Serum Albumin Human genes 0.000 description 2
- 108091006905 Human Serum Albumin Proteins 0.000 description 2
- 108060003951 Immunoglobulin Proteins 0.000 description 2
- 102000003746 Insulin Receptor Human genes 0.000 description 2
- 108010001127 Insulin Receptor Proteins 0.000 description 2
- 102100026720 Interferon beta Human genes 0.000 description 2
- 102100037850 Interferon gamma Human genes 0.000 description 2
- 108010047761 Interferon-alpha Proteins 0.000 description 2
- 102000006992 Interferon-alpha Human genes 0.000 description 2
- 108090000467 Interferon-beta Proteins 0.000 description 2
- 108010074328 Interferon-gamma Proteins 0.000 description 2
- 108090000177 Interleukin-11 Proteins 0.000 description 2
- 102000003815 Interleukin-11 Human genes 0.000 description 2
- 108090000978 Interleukin-4 Proteins 0.000 description 2
- 102000004388 Interleukin-4 Human genes 0.000 description 2
- 108010002616 Interleukin-5 Proteins 0.000 description 2
- 102000000743 Interleukin-5 Human genes 0.000 description 2
- 108090001005 Interleukin-6 Proteins 0.000 description 2
- 102000004889 Interleukin-6 Human genes 0.000 description 2
- 108010002586 Interleukin-7 Proteins 0.000 description 2
- 102000000704 Interleukin-7 Human genes 0.000 description 2
- 108010002335 Interleukin-9 Proteins 0.000 description 2
- 102000000585 Interleukin-9 Human genes 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 2
- AGPKZVBTJJNPAG-WHFBIAKZSA-N L-isoleucine Chemical compound CC[C@H](C)[C@H](N)C(O)=O AGPKZVBTJJNPAG-WHFBIAKZSA-N 0.000 description 2
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 2
- LRQKBLKVPFOOQJ-YFKPBYRVSA-N L-norleucine Chemical compound CCCC[C@H]([NH3+])C([O-])=O LRQKBLKVPFOOQJ-YFKPBYRVSA-N 0.000 description 2
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 2
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 2
- 108010063045 Lactoferrin Proteins 0.000 description 2
- 102000010445 Lactoferrin Human genes 0.000 description 2
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 2
- 239000005089 Luciferase Substances 0.000 description 2
- 108060001084 Luciferase Proteins 0.000 description 2
- 102000004083 Lymphotoxin-alpha Human genes 0.000 description 2
- 108090000542 Lymphotoxin-alpha Proteins 0.000 description 2
- 102100027754 Mast/stem cell growth factor receptor Kit Human genes 0.000 description 2
- 241000203407 Methanocaldococcus jannaschii Species 0.000 description 2
- 101710151805 Mitochondrial intermediate peptidase 1 Proteins 0.000 description 2
- 101000686985 Mouse mammary tumor virus (strain C3H) Protein PR73 Proteins 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 108010025020 Nerve Growth Factor Proteins 0.000 description 2
- 102100021584 Neurturin Human genes 0.000 description 2
- 108010015406 Neurturin Proteins 0.000 description 2
- 102000004140 Oncostatin M Human genes 0.000 description 2
- 108090000630 Oncostatin M Proteins 0.000 description 2
- 108091093037 Peptide nucleic acid Proteins 0.000 description 2
- 102100036154 Platelet basic protein Human genes 0.000 description 2
- 102100030304 Platelet factor 4 Human genes 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 229920002873 Polyethylenimine Polymers 0.000 description 2
- 101710118538 Protease Proteins 0.000 description 2
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 2
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 2
- 102000009572 RNA Polymerase II Human genes 0.000 description 2
- 108010009460 RNA Polymerase II Proteins 0.000 description 2
- 102000014450 RNA Polymerase III Human genes 0.000 description 2
- 108010078067 RNA Polymerase III Proteins 0.000 description 2
- 108091028664 Ribonucleotide Proteins 0.000 description 2
- 102100023361 SAP domain-containing ribonucleoprotein Human genes 0.000 description 2
- 102000018673 SEC Translocation Channels Human genes 0.000 description 2
- 108010091732 SEC Translocation Channels Proteins 0.000 description 2
- 101710194492 SET-binding protein Proteins 0.000 description 2
- 108091003202 SecA Proteins Proteins 0.000 description 2
- 206010040070 Septic Shock Diseases 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- 108010056088 Somatostatin Proteins 0.000 description 2
- 102000005157 Somatostatin Human genes 0.000 description 2
- 101000882406 Staphylococcus aureus Enterotoxin type C-1 Proteins 0.000 description 2
- 101000882403 Staphylococcus aureus Enterotoxin type C-2 Proteins 0.000 description 2
- 101001057112 Staphylococcus aureus Enterotoxin type D Proteins 0.000 description 2
- 108010090804 Streptavidin Proteins 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 102100030529 Suppressor of cytokine signaling 7 Human genes 0.000 description 2
- 108010078233 Thymalfasin Proteins 0.000 description 2
- 102400000800 Thymosin alpha-1 Human genes 0.000 description 2
- 206010044248 Toxic shock syndrome Diseases 0.000 description 2
- 231100000650 Toxic shock syndrome Toxicity 0.000 description 2
- 101710120037 Toxin CcdB Proteins 0.000 description 2
- 102000040945 Transcription factor Human genes 0.000 description 2
- 108091023040 Transcription factor Proteins 0.000 description 2
- 102100040247 Tumor necrosis factor Human genes 0.000 description 2
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 description 2
- 108090000435 Urokinase-type plasminogen activator Proteins 0.000 description 2
- 102000003990 Urokinase-type plasminogen activator Human genes 0.000 description 2
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 2
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 125000002252 acyl group Chemical group 0.000 description 2
- 125000003342 alkenyl group Chemical group 0.000 description 2
- 125000000217 alkyl group Chemical group 0.000 description 2
- 102000015395 alpha 1-Antitrypsin Human genes 0.000 description 2
- 108010050122 alpha 1-Antitrypsin Proteins 0.000 description 2
- 229940024142 alpha 1-antitrypsin Drugs 0.000 description 2
- 125000003277 amino group Chemical group 0.000 description 2
- 229960000723 ampicillin Drugs 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
- 230000002587 anti-hemolytic effect Effects 0.000 description 2
- 230000000692 anti-sense effect Effects 0.000 description 2
- 108010082685 antiarrhythmic peptide Proteins 0.000 description 2
- 239000002518 antifoaming agent Substances 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 210000004436 artificial bacterial chromosome Anatomy 0.000 description 2
- 210000001106 artificial yeast chromosome Anatomy 0.000 description 2
- 125000003118 aryl group Chemical group 0.000 description 2
- 235000009582 asparagine Nutrition 0.000 description 2
- 229960001230 asparagine Drugs 0.000 description 2
- FZCSTZYAHCUGEM-UHFFFAOYSA-N aspergillomarasmine B Natural products OC(=O)CNC(C(O)=O)CNC(C(O)=O)CC(O)=O FZCSTZYAHCUGEM-UHFFFAOYSA-N 0.000 description 2
- 125000004429 atom Chemical group 0.000 description 2
- 150000001540 azides Chemical class 0.000 description 2
- 239000007640 basal medium Substances 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 229960002685 biotin Drugs 0.000 description 2
- 235000020958 biotin Nutrition 0.000 description 2
- 239000011616 biotin Substances 0.000 description 2
- 150000001615 biotins Chemical class 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- ZADPBFCGQRWHPN-UHFFFAOYSA-N boronic acid Chemical compound OBO ZADPBFCGQRWHPN-UHFFFAOYSA-N 0.000 description 2
- 108091006374 cAMP receptor proteins Proteins 0.000 description 2
- BBBFJLBPOGFECG-VJVYQDLKSA-N calcitonin Chemical compound N([C@H](C(=O)N[C@@H](CC(C)C)C(=O)NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1NC=NC=1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(N)=O)C(C)C)C(=O)[C@@H]1CSSC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1 BBBFJLBPOGFECG-VJVYQDLKSA-N 0.000 description 2
- 229960004015 calcitonin Drugs 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 125000003178 carboxy group Chemical group [H]OC(*)=O 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
- 230000015556 catabolic process Effects 0.000 description 2
- 230000036978 cell physiology Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 229960005091 chloramphenicol Drugs 0.000 description 2
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 239000013611 chromosomal DNA Substances 0.000 description 2
- 229920001436 collagen Polymers 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- UFULAYFCSOUIOV-UHFFFAOYSA-N cysteamine Chemical compound NCCS UFULAYFCSOUIOV-UHFFFAOYSA-N 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 2
- 239000003623 enhancer Substances 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 230000029142 excretion Effects 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 229960004222 factor ix Drugs 0.000 description 2
- 229940012413 factor vii Drugs 0.000 description 2
- 229940012952 fibrinogen Drugs 0.000 description 2
- 125000000524 functional group Chemical group 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 125000000404 glutamine group Chemical group N[C@@H](CCC(N)=O)C(=O)* 0.000 description 2
- YPZRWBKMTBYPTK-BJDJZHNGSA-N glutathione disulfide Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@H](C(=O)NCC(O)=O)CSSC[C@@H](C(=O)NCC(O)=O)NC(=O)CC[C@H](N)C(O)=O YPZRWBKMTBYPTK-BJDJZHNGSA-N 0.000 description 2
- 230000013595 glycosylation Effects 0.000 description 2
- 238000006206 glycosylation reaction Methods 0.000 description 2
- 239000002622 gonadotropin Substances 0.000 description 2
- 210000003714 granulocyte Anatomy 0.000 description 2
- 125000001475 halogen functional group Chemical group 0.000 description 2
- WQPDUTSPKFMPDP-OUMQNGNKSA-N hirudin Chemical compound C([C@@H](C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC(OS(O)(=O)=O)=CC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCCN)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H]1NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H]2CSSC[C@@H](C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(=O)N[C@H](C(NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N2)=O)CSSC1)C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=2C=CC(O)=CC=2)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)C(C)C)[C@@H](C)O)CSSC1)C(C)C)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 WQPDUTSPKFMPDP-OUMQNGNKSA-N 0.000 description 2
- 229940006607 hirudin Drugs 0.000 description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 2
- 101150035627 hoc gene Proteins 0.000 description 2
- 229930195733 hydrocarbon Natural products 0.000 description 2
- 150000002430 hydrocarbons Chemical class 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 150000002466 imines Chemical class 0.000 description 2
- 102000018358 immunoglobulin Human genes 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 230000000977 initiatory effect Effects 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 229940079322 interferon Drugs 0.000 description 2
- 229940047124 interferons Drugs 0.000 description 2
- 229940047122 interleukins Drugs 0.000 description 2
- 239000002563 ionic surfactant Substances 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 229960000310 isoleucine Drugs 0.000 description 2
- AGPKZVBTJJNPAG-UHFFFAOYSA-N isoleucine Natural products CCC(C)C(N)C(O)=O AGPKZVBTJJNPAG-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 229940078795 lactoferrin Drugs 0.000 description 2
- 235000021242 lactoferrin Nutrition 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 230000004807 localization Effects 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 210000002540 macrophage Anatomy 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 238000002703 mutagenesis Methods 0.000 description 2
- 231100000350 mutagenesis Toxicity 0.000 description 2
- OHDXDNUPVVYWOV-UHFFFAOYSA-N n-methyl-1-(2-naphthalen-1-ylsulfanylphenyl)methanamine Chemical compound CNCC1=CC=CC=C1SC1=CC=CC2=CC=CC=C12 OHDXDNUPVVYWOV-UHFFFAOYSA-N 0.000 description 2
- 238000010899 nucleation Methods 0.000 description 2
- 230000000269 nucleophilic effect Effects 0.000 description 2
- 238000002515 oligonucleotide synthesis Methods 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 230000002018 overexpression Effects 0.000 description 2
- 230000001590 oxidative effect Effects 0.000 description 2
- PSWJVKKJYCAPTI-UHFFFAOYSA-N oxido-oxo-phosphonophosphanylphosphanium Chemical compound OP(O)(=O)PP(=O)=O PSWJVKKJYCAPTI-UHFFFAOYSA-N 0.000 description 2
- 239000000199 parathyroid hormone Substances 0.000 description 2
- 150000002993 phenylalanine derivatives Chemical class 0.000 description 2
- 102000005162 pleiotrophin Human genes 0.000 description 2
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 101150080066 proS1 gene Proteins 0.000 description 2
- 230000009465 prokaryotic expression Effects 0.000 description 2
- 230000030788 protein refolding Effects 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 238000009790 rate-determining step (RDS) Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 239000002336 ribonucleotide Substances 0.000 description 2
- 125000002652 ribonucleotide group Chemical group 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000001488 sodium phosphate Substances 0.000 description 2
- 229910000162 sodium phosphate Inorganic materials 0.000 description 2
- NHXLMOGPVYXJNR-ATOGVRKGSA-N somatostatin Chemical compound C([C@H]1C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CSSC[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2C3=CC=CC=C3NC=2)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(=O)N1)[C@@H](C)O)NC(=O)CNC(=O)[C@H](C)N)C(O)=O)=O)[C@H](O)C)C1=CC=CC=C1 NHXLMOGPVYXJNR-ATOGVRKGSA-N 0.000 description 2
- 229960000553 somatostatin Drugs 0.000 description 2
- 150000003431 steroids Chemical class 0.000 description 2
- 231100000617 superantigen Toxicity 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 125000003396 thiol group Chemical group [H]S* 0.000 description 2
- RYYWUUFWQRZTIU-UHFFFAOYSA-K thiophosphate Chemical compound [O-]P([O-])([O-])=S RYYWUUFWQRZTIU-UHFFFAOYSA-K 0.000 description 2
- NZVYCXVTEHPMHE-ZSUJOUNUSA-N thymalfasin Chemical compound CC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O NZVYCXVTEHPMHE-ZSUJOUNUSA-N 0.000 description 2
- 229960004231 thymalfasin Drugs 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 2
- 229960005356 urokinase Drugs 0.000 description 2
- 239000004474 valine Substances 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- 239000002888 zwitterionic surfactant Substances 0.000 description 2
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 2
- VFCYASLQFGXHTQ-VIFPVBQESA-N (2s)-2-[benzyl(nitro)amino]-3-hydroxypropanoic acid Chemical compound OC[C@@H](C(O)=O)N([N+]([O-])=O)CC1=CC=CC=C1 VFCYASLQFGXHTQ-VIFPVBQESA-N 0.000 description 1
- LJHYWUVYIKCPGU-VIFPVBQESA-N (2s)-2-amino-3-[4-(carboxymethyl)phenyl]propanoic acid Chemical compound OC(=O)[C@@H](N)CC1=CC=C(CC(O)=O)C=C1 LJHYWUVYIKCPGU-VIFPVBQESA-N 0.000 description 1
- NLFOHNAFILVHGM-AWEZNQCLSA-N (2s)-2-amino-3-[4-[(2-nitrophenyl)methoxy]phenyl]propanoic acid Chemical compound C1=CC(C[C@H](N)C(O)=O)=CC=C1OCC1=CC=CC=C1[N+]([O-])=O NLFOHNAFILVHGM-AWEZNQCLSA-N 0.000 description 1
- GTVVZTAFGPQSPC-QMMMGPOBSA-N (2s)-2-azaniumyl-3-(4-nitrophenyl)propanoate Chemical compound OC(=O)[C@@H](N)CC1=CC=C([N+]([O-])=O)C=C1 GTVVZTAFGPQSPC-QMMMGPOBSA-N 0.000 description 1
- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical compound C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 description 1
- YPGMOWHXEQDBBV-QWWZWVQMSA-N (4S,5S)-1,2-dithiane-4,5-diol Chemical compound O[C@@H]1CSSC[C@H]1O YPGMOWHXEQDBBV-QWWZWVQMSA-N 0.000 description 1
- 108010052418 (N-(2-((4-((2-((4-(9-acridinylamino)phenyl)amino)-2-oxoethyl)amino)-4-oxobutyl)amino)-1-(1H-imidazol-4-ylmethyl)-1-oxoethyl)-6-(((-2-aminoethyl)amino)methyl)-2-pyridinecarboxamidato) iron(1+) Proteins 0.000 description 1
- WHTVZRBIWZFKQO-AWEZNQCLSA-N (S)-chloroquine Chemical compound ClC1=CC=C2C(N[C@@H](C)CCCN(CC)CC)=CC=NC2=C1 WHTVZRBIWZFKQO-AWEZNQCLSA-N 0.000 description 1
- PSBDWGZCVUAZQS-UHFFFAOYSA-N (dimethylsulfonio)acetate Chemical compound C[S+](C)CC([O-])=O PSBDWGZCVUAZQS-UHFFFAOYSA-N 0.000 description 1
- UKAUYVFTDYCKQA-UHFFFAOYSA-N -2-Amino-4-hydroxybutanoic acid Natural products OC(=O)C(N)CCO UKAUYVFTDYCKQA-UHFFFAOYSA-N 0.000 description 1
- TZCPCKNHXULUIY-RGULYWFUSA-N 1,2-distearoyl-sn-glycero-3-phosphoserine Chemical compound CCCCCCCCCCCCCCCCCC(=O)OC[C@H](COP(O)(=O)OC[C@H](N)C(O)=O)OC(=O)CCCCCCCCCCCCCCCCC TZCPCKNHXULUIY-RGULYWFUSA-N 0.000 description 1
- 102100024341 10 kDa heat shock protein, mitochondrial Human genes 0.000 description 1
- VGONTNSXDCQUGY-RRKCRQDMSA-N 2'-deoxyinosine Chemical group C1[C@H](O)[C@@H](CO)O[C@H]1N1C(N=CNC2=O)=C2N=C1 VGONTNSXDCQUGY-RRKCRQDMSA-N 0.000 description 1
- SMZOUWXMTYCWNB-UHFFFAOYSA-N 2-(2-methoxy-5-methylphenyl)ethanamine Chemical compound COC1=CC=C(C)C=C1CCN SMZOUWXMTYCWNB-UHFFFAOYSA-N 0.000 description 1
- NIXOWILDQLNWCW-UHFFFAOYSA-N 2-Propenoic acid Natural products OC(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 description 1
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 description 1
- UMCMPZBLKLEWAF-BCTGSCMUSA-N 3-[(3-cholamidopropyl)dimethylammonio]propane-1-sulfonate Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(=O)NCCC[N+](C)(C)CCCS([O-])(=O)=O)C)[C@@]2(C)[C@@H](O)C1 UMCMPZBLKLEWAF-BCTGSCMUSA-N 0.000 description 1
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 description 1
- UQTZMGFTRHFAAM-ZETCQYMHSA-N 3-iodo-L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C(I)=C1 UQTZMGFTRHFAAM-ZETCQYMHSA-N 0.000 description 1
- FBTSQILOGYXGMD-LURJTMIESA-N 3-nitro-L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C([N+]([O-])=O)=C1 FBTSQILOGYXGMD-LURJTMIESA-N 0.000 description 1
- HLXHCNWEVQNNKA-UHFFFAOYSA-N 5-methoxy-2,3-dihydro-1h-inden-2-amine Chemical compound COC1=CC=C2CC(N)CC2=C1 HLXHCNWEVQNNKA-UHFFFAOYSA-N 0.000 description 1
- 102100038222 60 kDa heat shock protein, mitochondrial Human genes 0.000 description 1
- CJIJXIFQYOPWTF-UHFFFAOYSA-N 7-hydroxycoumarin Natural products O1C(=O)C=CC2=CC(O)=CC=C21 CJIJXIFQYOPWTF-UHFFFAOYSA-N 0.000 description 1
- HBAQYPYDRFILMT-UHFFFAOYSA-N 8-[3-(1-cyclopropylpyrazol-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl]-3-methyl-3,8-diazabicyclo[3.2.1]octan-2-one Chemical class C1(CC1)N1N=CC(=C1)C1=NNC2=C1N=C(N=C2)N1C2C(N(CC1CC2)C)=O HBAQYPYDRFILMT-UHFFFAOYSA-N 0.000 description 1
- 108091005508 Acid proteases Proteins 0.000 description 1
- 241000567139 Aeropyrum pernix Species 0.000 description 1
- 108700028369 Alleles Proteins 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 239000004382 Amylase Substances 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- 102000000412 Annexin Human genes 0.000 description 1
- 108050008874 Annexin Proteins 0.000 description 1
- 108020005544 Antisense RNA Proteins 0.000 description 1
- 102000007592 Apolipoproteins Human genes 0.000 description 1
- 108010071619 Apolipoproteins Proteins 0.000 description 1
- OZDNDGXASTWERN-CTNGQTDRSA-N Apovincamine Chemical compound C1=CC=C2C(CCN3CCC4)=C5[C@@H]3[C@]4(CC)C=C(C(=O)OC)N5C2=C1 OZDNDGXASTWERN-CTNGQTDRSA-N 0.000 description 1
- 101100350628 Arabidopsis thaliana PLL3 gene Proteins 0.000 description 1
- 101000634115 Arabidopsis thaliana RNA polymerase sigma factor sigE, chloroplastic/mitochondrial Proteins 0.000 description 1
- 241000205042 Archaeoglobus fulgidus Species 0.000 description 1
- 101100425074 Aspergillus oryzae (strain ATCC 42149 / RIB 40) thiA gene Proteins 0.000 description 1
- 101150071434 BAR1 gene Proteins 0.000 description 1
- 108020004513 Bacterial RNA Proteins 0.000 description 1
- 108091032955 Bacterial small RNA Proteins 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- 102000004219 Brain-derived neurotrophic factor Human genes 0.000 description 1
- 108090000715 Brain-derived neurotrophic factor Proteins 0.000 description 1
- 102100021943 C-C motif chemokine 2 Human genes 0.000 description 1
- 101710155857 C-C motif chemokine 2 Proteins 0.000 description 1
- 101710167766 C-type lectin domain family 11 member A Proteins 0.000 description 1
- 102100032528 C-type lectin domain family 11 member A Human genes 0.000 description 1
- 108050006947 CXC Chemokine Proteins 0.000 description 1
- 102000019388 CXC chemokine Human genes 0.000 description 1
- 101100504320 Caenorhabditis elegans mcp-1 gene Proteins 0.000 description 1
- KXDHJXZQYSOELW-UHFFFAOYSA-M Carbamate Chemical compound NC([O-])=O KXDHJXZQYSOELW-UHFFFAOYSA-M 0.000 description 1
- 108010006303 Carboxypeptidases Proteins 0.000 description 1
- 102000005367 Carboxypeptidases Human genes 0.000 description 1
- 102000014914 Carrier Proteins Human genes 0.000 description 1
- 241000218645 Cedrus Species 0.000 description 1
- 206010057248 Cell death Diseases 0.000 description 1
- 101710117022 Chaperone protein HscA Proteins 0.000 description 1
- 108010059013 Chaperonin 10 Proteins 0.000 description 1
- 108010058432 Chaperonin 60 Proteins 0.000 description 1
- 241001112695 Clostridiales Species 0.000 description 1
- 239000004971 Cross linker Substances 0.000 description 1
- 241000192700 Cyanobacteria Species 0.000 description 1
- LEVWYRKDKASIDU-QWWZWVQMSA-N D-cystine Chemical compound OC(=O)[C@H](N)CSSC[C@@H](N)C(O)=O LEVWYRKDKASIDU-QWWZWVQMSA-N 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- QRLVDLBMBULFAL-UHFFFAOYSA-N Digitonin Natural products CC1CCC2(OC1)OC3C(O)C4C5CCC6CC(OC7OC(CO)C(OC8OC(CO)C(O)C(OC9OCC(O)C(O)C9OC%10OC(CO)C(O)C(OC%11OC(CO)C(O)C(O)C%11O)C%10O)C8O)C(O)C7O)C(O)CC6(C)C5CCC4(C)C3C2C QRLVDLBMBULFAL-UHFFFAOYSA-N 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 238000012286 ELISA Assay Methods 0.000 description 1
- 229940122858 Elastase inhibitor Drugs 0.000 description 1
- 101100409165 Escherichia coli (strain K12) prc gene Proteins 0.000 description 1
- 101000867232 Escherichia coli Heat-stable enterotoxin II Proteins 0.000 description 1
- 101100332643 Escherichia coli O78:H11 (strain H10407 / ETEC) eatA gene Proteins 0.000 description 1
- 241000701959 Escherichia virus Lambda Species 0.000 description 1
- 108060002716 Exonuclease Proteins 0.000 description 1
- 108010014172 Factor V Proteins 0.000 description 1
- 108010054218 Factor VIII Proteins 0.000 description 1
- 102000001690 Factor VIII Human genes 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108700007698 Genetic Terminator Regions Proteins 0.000 description 1
- 241000193385 Geobacillus stearothermophilus Species 0.000 description 1
- 102000034615 Glial cell line-derived neurotrophic factor Human genes 0.000 description 1
- 108091010837 Glial cell line-derived neurotrophic factor Proteins 0.000 description 1
- 108010017544 Glucosylceramidase Proteins 0.000 description 1
- 102000004547 Glucosylceramidase Human genes 0.000 description 1
- 108010053070 Glutathione Disulfide Proteins 0.000 description 1
- JZNWSCPGTDBMEW-UHFFFAOYSA-N Glycerophosphorylethanolamin Natural products NCCOP(O)(=O)OCC(O)CO JZNWSCPGTDBMEW-UHFFFAOYSA-N 0.000 description 1
- ZWZWYGMENQVNFU-UHFFFAOYSA-N Glycerophosphorylserin Natural products OC(=O)C(N)COP(O)(=O)OCC(O)CO ZWZWYGMENQVNFU-UHFFFAOYSA-N 0.000 description 1
- 229930186217 Glycolipid Natural products 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 102100023737 GrpE protein homolog 1, mitochondrial Human genes 0.000 description 1
- 229940121710 HMGCoA reductase inhibitor Drugs 0.000 description 1
- 102000004447 HSP40 Heat-Shock Proteins Human genes 0.000 description 1
- 108010042283 HSP40 Heat-Shock Proteins Proteins 0.000 description 1
- 241000204933 Haloferax volcanii Species 0.000 description 1
- 108090000100 Hepatocyte Growth Factor Proteins 0.000 description 1
- 102100021866 Hepatocyte growth factor Human genes 0.000 description 1
- 101000938776 Homo sapiens ETS domain-containing transcription factor ERF Proteins 0.000 description 1
- 101000829489 Homo sapiens GrpE protein homolog 1, mitochondrial Proteins 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 206010021143 Hypoxia Diseases 0.000 description 1
- 108700005091 Immunoglobulin Genes Proteins 0.000 description 1
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 description 1
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 description 1
- 102000013463 Immunoglobulin Light Chains Human genes 0.000 description 1
- 108010065825 Immunoglobulin Light Chains Proteins 0.000 description 1
- 229940118432 Interleukin receptor antagonist Drugs 0.000 description 1
- 108020004684 Internal Ribosome Entry Sites Proteins 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- SRBFZHDQGSBBOR-HWQSCIPKSA-N L-arabinopyranose Chemical compound O[C@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-HWQSCIPKSA-N 0.000 description 1
- UKAUYVFTDYCKQA-VKHMYHEASA-N L-homoserine Chemical compound OC(=O)[C@@H](N)CCO UKAUYVFTDYCKQA-VKHMYHEASA-N 0.000 description 1
- QEFRNWWLZKMPFJ-ZXPFJRLXSA-N L-methionine (R)-S-oxide Chemical compound C[S@@](=O)CC[C@H]([NH3+])C([O-])=O QEFRNWWLZKMPFJ-ZXPFJRLXSA-N 0.000 description 1
- QEFRNWWLZKMPFJ-UHFFFAOYSA-N L-methionine sulphoxide Natural products CS(=O)CCC(N)C(O)=O QEFRNWWLZKMPFJ-UHFFFAOYSA-N 0.000 description 1
- ZFOMKMMPBOQKMC-KXUCPTDWSA-N L-pyrrolysine Chemical compound C[C@@H]1CC=N[C@H]1C(=O)NCCCC[C@H]([NH3+])C([O-])=O ZFOMKMMPBOQKMC-KXUCPTDWSA-N 0.000 description 1
- 102000000853 LDL receptors Human genes 0.000 description 1
- 239000006142 Luria-Bertani Agar Substances 0.000 description 1
- 239000006137 Luria-Bertani broth Substances 0.000 description 1
- 241000064099 Massilioclostridium coli Species 0.000 description 1
- 241001302042 Methanothermobacter thermautotrophicus Species 0.000 description 1
- 241000191938 Micrococcus luteus Species 0.000 description 1
- 241001430197 Mollicutes Species 0.000 description 1
- CHJJGSNFBQVOTG-UHFFFAOYSA-N N-methyl-guanidine Natural products CNC(N)=N CHJJGSNFBQVOTG-UHFFFAOYSA-N 0.000 description 1
- 102000015336 Nerve Growth Factor Human genes 0.000 description 1
- 102000007072 Nerve Growth Factors Human genes 0.000 description 1
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 1
- 238000000636 Northern blotting Methods 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 108700026244 Open Reading Frames Proteins 0.000 description 1
- 101710116435 Outer membrane protein Proteins 0.000 description 1
- 101100378536 Ovis aries ADRB1 gene Proteins 0.000 description 1
- 101150053185 P450 gene Proteins 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 102000003982 Parathyroid hormone Human genes 0.000 description 1
- 108090000445 Parathyroid hormone Proteins 0.000 description 1
- 108010090127 Periplasmic Proteins Proteins 0.000 description 1
- 108090000316 Pitrilysin Proteins 0.000 description 1
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 description 1
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 description 1
- 102000029797 Prion Human genes 0.000 description 1
- 108091000054 Prion Proteins 0.000 description 1
- 108010076181 Proinsulin Proteins 0.000 description 1
- 101800004937 Protein C Proteins 0.000 description 1
- 102000017975 Protein C Human genes 0.000 description 1
- 102000001253 Protein Kinase Human genes 0.000 description 1
- 102000016971 Proto-Oncogene Proteins c-kit Human genes 0.000 description 1
- 108010014608 Proto-Oncogene Proteins c-kit Proteins 0.000 description 1
- 241000205160 Pyrococcus Species 0.000 description 1
- 241000205156 Pyrococcus furiosus Species 0.000 description 1
- 241000522615 Pyrococcus horikoshii Species 0.000 description 1
- ASNFTDCKZKHJSW-REOHCLBHSA-N Quisqualic acid Chemical class OC(=O)[C@@H](N)CN1OC(=O)NC1=O ASNFTDCKZKHJSW-REOHCLBHSA-N 0.000 description 1
- 102000017143 RNA Polymerase I Human genes 0.000 description 1
- 108010013845 RNA Polymerase I Proteins 0.000 description 1
- 230000006819 RNA synthesis Effects 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 108010034634 Repressor Proteins Proteins 0.000 description 1
- 101100319895 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) YAP3 gene Proteins 0.000 description 1
- 101100160515 Saccharomyces cerevisiae (strain ATCC 204508 / S288c) YPS1 gene Proteins 0.000 description 1
- 241000831652 Salinivibrio sharmensis Species 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 101800001700 Saposin-D Proteins 0.000 description 1
- 108010034546 Serratia marcescens nuclease Proteins 0.000 description 1
- 102000007562 Serum Albumin Human genes 0.000 description 1
- 108010071390 Serum Albumin Proteins 0.000 description 1
- 102100037253 Solute carrier family 45 member 3 Human genes 0.000 description 1
- 102100021669 Stromal cell-derived factor 1 Human genes 0.000 description 1
- 101710088580 Stromal cell-derived factor 1 Proteins 0.000 description 1
- 108700005078 Synthetic Genes Proteins 0.000 description 1
- 108700026226 TATA Box Proteins 0.000 description 1
- 241000589499 Thermus thermophilus Species 0.000 description 1
- RYYWUUFWQRZTIU-UHFFFAOYSA-N Thiophosphoric acid Chemical class OP(O)(S)=O RYYWUUFWQRZTIU-UHFFFAOYSA-N 0.000 description 1
- 108700009124 Transcription Initiation Site Proteins 0.000 description 1
- 229940122618 Trypsin inhibitor Drugs 0.000 description 1
- 101710162629 Trypsin inhibitor Proteins 0.000 description 1
- 102100034298 Tyrosine-tRNA ligase, cytoplasmic Human genes 0.000 description 1
- IXKSXJFAGXLQOQ-XISFHERQSA-N WHWLQLKPGQPMY Chemical group C([C@@H](C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)NC(=O)[C@@H](N)CC=1C2=CC=CC=C2NC=1)C1=CNC=N1 IXKSXJFAGXLQOQ-XISFHERQSA-N 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 125000002339 acetoacetyl group Chemical group O=C([*])C([H])([H])C(=O)C([H])([H])[H] 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000009056 active transport Effects 0.000 description 1
- 230000010933 acylation Effects 0.000 description 1
- 238000005917 acylation reaction Methods 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 239000002168 alkylating agent Substances 0.000 description 1
- 229940100198 alkylating agent Drugs 0.000 description 1
- 125000000304 alkynyl group Chemical group 0.000 description 1
- 150000001371 alpha-amino acids Chemical class 0.000 description 1
- 235000008206 alpha-amino acids Nutrition 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 230000006229 amino acid addition Effects 0.000 description 1
- 229940093740 amino acid and derivative Drugs 0.000 description 1
- 239000000908 ammonium hydroxide Substances 0.000 description 1
- 101150073130 ampR gene Proteins 0.000 description 1
- 235000019418 amylase Nutrition 0.000 description 1
- 230000001195 anabolic effect Effects 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 239000000935 antidepressant agent Substances 0.000 description 1
- 229940005513 antidepressants Drugs 0.000 description 1
- 229940121375 antifungal agent Drugs 0.000 description 1
- 239000003429 antifungal agent Substances 0.000 description 1
- 239000003430 antimalarial agent Substances 0.000 description 1
- 229940033495 antimalarials Drugs 0.000 description 1
- OZDNDGXASTWERN-UHFFFAOYSA-N apovincamine Natural products C1=CC=C2C(CCN3CCC4)=C5C3C4(CC)C=C(C(=O)OC)N5C2=C1 OZDNDGXASTWERN-UHFFFAOYSA-N 0.000 description 1
- 230000011681 asexual reproduction Effects 0.000 description 1
- 238000013465 asexual reproduction Methods 0.000 description 1
- 208000027697 autoimmune lymphoproliferative syndrome due to CTLA4 haploinsuffiency Diseases 0.000 description 1
- 210000003578 bacterial chromosome Anatomy 0.000 description 1
- 239000003899 bactericide agent Substances 0.000 description 1
- 238000007630 basic procedure Methods 0.000 description 1
- 125000003236 benzoyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C(*)=O 0.000 description 1
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 1
- 150000001576 beta-amino acids Chemical class 0.000 description 1
- 239000003613 bile acid Substances 0.000 description 1
- 108091008324 binding proteins Proteins 0.000 description 1
- 238000010364 biochemical engineering Methods 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 229940088623 biologically active substance Drugs 0.000 description 1
- 230000001851 biosynthetic effect Effects 0.000 description 1
- JCZLABDVDPYLRZ-AWEZNQCLSA-N biphenylalanine Chemical compound C1=CC(C[C@H](N)C(O)=O)=CC=C1C1=CC=CC=C1 JCZLABDVDPYLRZ-AWEZNQCLSA-N 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 229940019700 blood coagulation factors Drugs 0.000 description 1
- 229940077737 brain-derived neurotrophic factor Drugs 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 238000005251 capillar electrophoresis Methods 0.000 description 1
- 150000001719 carbohydrate derivatives Chemical class 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 229940125692 cardiovascular agent Drugs 0.000 description 1
- 239000002327 cardiovascular agent Substances 0.000 description 1
- 238000005341 cation exchange Methods 0.000 description 1
- 239000003093 cationic surfactant Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 238000009614 chemical analysis method Methods 0.000 description 1
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 1
- 229960003677 chloroquine Drugs 0.000 description 1
- WHTVZRBIWZFKQO-UHFFFAOYSA-N chloroquine Natural products ClC1=CC=C2C(NC(C)CCCN(CC)CC)=CC=NC2=C1 WHTVZRBIWZFKQO-UHFFFAOYSA-N 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 101150036359 clpB gene Proteins 0.000 description 1
- 101150038575 clpS gene Proteins 0.000 description 1
- 101150096566 clpX gene Proteins 0.000 description 1
- 230000035071 co-translational protein modification Effects 0.000 description 1
- 239000003184 complementary RNA Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 239000002872 contrast media Substances 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 108010082242 corticosterone receptor Proteins 0.000 description 1
- 238000004132 cross linking Methods 0.000 description 1
- 239000003431 cross linking reagent Substances 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 125000004093 cyano group Chemical group *C#N 0.000 description 1
- OOTFVKOQINZBBF-UHFFFAOYSA-N cystamine Chemical compound CCSSCCN OOTFVKOQINZBBF-UHFFFAOYSA-N 0.000 description 1
- 229940099500 cystamine Drugs 0.000 description 1
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 1
- 229960003067 cystine Drugs 0.000 description 1
- 210000000172 cytosol Anatomy 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 239000002254 cytotoxic agent Substances 0.000 description 1
- 229940127089 cytotoxic agent Drugs 0.000 description 1
- 231100000599 cytotoxic agent Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000006324 decarbonylation Effects 0.000 description 1
- 238000006606 decarbonylation reaction Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 229960003964 deoxycholic acid Drugs 0.000 description 1
- 239000005549 deoxyribonucleoside Substances 0.000 description 1
- 239000005547 deoxyribonucleotide Substances 0.000 description 1
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000000032 diagnostic agent Substances 0.000 description 1
- 229940039227 diagnostic agent Drugs 0.000 description 1
- 125000001664 diethylamino group Chemical group [H]C([H])([H])C([H])([H])N(*)C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- UVYVLBIGDKGWPX-KUAJCENISA-N digitonin Chemical compound O([C@@H]1[C@@H]([C@]2(CC[C@@H]3[C@@]4(C)C[C@@H](O)[C@H](O[C@H]5[C@@H]([C@@H](O)[C@@H](O[C@H]6[C@@H]([C@@H](O[C@H]7[C@@H]([C@@H](O)[C@H](O)CO7)O)[C@H](O)[C@@H](CO)O6)O[C@H]6[C@@H]([C@@H](O[C@H]7[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O7)O)[C@@H](O)[C@@H](CO)O6)O)[C@@H](CO)O5)O)C[C@@H]4CC[C@H]3[C@@H]2[C@@H]1O)C)[C@@H]1C)[C@]11CC[C@@H](C)CO1 UVYVLBIGDKGWPX-KUAJCENISA-N 0.000 description 1
- UVYVLBIGDKGWPX-UHFFFAOYSA-N digitonine Natural products CC1C(C2(CCC3C4(C)CC(O)C(OC5C(C(O)C(OC6C(C(OC7C(C(O)C(O)CO7)O)C(O)C(CO)O6)OC6C(C(OC7C(C(O)C(O)C(CO)O7)O)C(O)C(CO)O6)O)C(CO)O5)O)CC4CCC3C2C2O)C)C2OC11CCC(C)CO1 UVYVLBIGDKGWPX-UHFFFAOYSA-N 0.000 description 1
- SWSQBOPZIKWTGO-UHFFFAOYSA-N dimethylaminoamidine Natural products CN(C)C(N)=N SWSQBOPZIKWTGO-UHFFFAOYSA-N 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 150000002019 disulfides Chemical class 0.000 description 1
- VHJLVAABSRFDPM-ZXZARUISSA-N dithioerythritol Chemical compound SC[C@H](O)[C@H](O)CS VHJLVAABSRFDPM-ZXZARUISSA-N 0.000 description 1
- NAGJZTKCGNOGPW-UHFFFAOYSA-N dithiophosphoric acid Chemical class OP(O)(S)=S NAGJZTKCGNOGPW-UHFFFAOYSA-N 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 239000003602 elastase inhibitor Substances 0.000 description 1
- 238000007350 electrophilic reaction Methods 0.000 description 1
- 230000008519 endogenous mechanism Effects 0.000 description 1
- 231100000655 enterotoxin Toxicity 0.000 description 1
- 230000007515 enzymatic degradation Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- 102000013165 exonuclease Human genes 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000010195 expression analysis Methods 0.000 description 1
- 238000013213 extrapolation Methods 0.000 description 1
- 229960000301 factor viii Drugs 0.000 description 1
- 229940012426 factor x Drugs 0.000 description 1
- 238000005187 foaming Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 229960002743 glutamine Drugs 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 229960004198 guanidine Drugs 0.000 description 1
- 229960000789 guanidine hydrochloride Drugs 0.000 description 1
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 125000000623 heterocyclic group Chemical group 0.000 description 1
- IPCSVZSSVZVIGE-UHFFFAOYSA-M hexadecanoate Chemical compound CCCCCCCCCCCCCCCC([O-])=O IPCSVZSSVZVIGE-UHFFFAOYSA-M 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 108091008039 hormone receptors Proteins 0.000 description 1
- 230000006658 host protein synthesis Effects 0.000 description 1
- 238000007849 hot-start PCR Methods 0.000 description 1
- 229940042795 hydrazides for tuberculosis treatment Drugs 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000002471 hydroxymethylglutaryl coenzyme A reductase inhibitor Substances 0.000 description 1
- 230000001146 hypoxic effect Effects 0.000 description 1
- 150000007976 iminium ions Chemical class 0.000 description 1
- 230000014726 immortalization of host cell Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 239000003262 industrial enzyme Substances 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000012212 insulator Substances 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 230000002361 ketogenic effect Effects 0.000 description 1
- 210000003127 knee Anatomy 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000006194 liquid suspension Substances 0.000 description 1
- 229940076783 lucentis Drugs 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 210000005060 membrane bound organelle Anatomy 0.000 description 1
- 230000004630 mental health Effects 0.000 description 1
- 229960003151 mercaptamine Drugs 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- YACKEPLHDIMKIO-UHFFFAOYSA-N methylphosphonic acid Chemical compound CP(O)(O)=O YACKEPLHDIMKIO-UHFFFAOYSA-N 0.000 description 1
- LSDPWZHWYPCBBB-UHFFFAOYSA-O methylsulfide anion Chemical compound [SH2+]C LSDPWZHWYPCBBB-UHFFFAOYSA-O 0.000 description 1
- 239000000693 micelle Substances 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 108091005601 modified peptides Proteins 0.000 description 1
- 239000003607 modifier Substances 0.000 description 1
- 230000002956 necrotizing effect Effects 0.000 description 1
- 230000001613 neoplastic effect Effects 0.000 description 1
- 229940053128 nerve growth factor Drugs 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 125000003835 nucleoside group Chemical group 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 101150008884 osmY gene Proteins 0.000 description 1
- 230000011164 ossification Effects 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- YPZRWBKMTBYPTK-UHFFFAOYSA-N oxidized gamma-L-glutamyl-L-cysteinylglycine Natural products OC(=O)C(N)CCC(=O)NC(C(=O)NCC(O)=O)CSSCC(C(=O)NCC(O)=O)NC(=O)CCC(N)C(O)=O YPZRWBKMTBYPTK-UHFFFAOYSA-N 0.000 description 1
- 230000026792 palmitoylation Effects 0.000 description 1
- 229960001319 parathyroid hormone Drugs 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 230000007030 peptide scission Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 description 1
- 150000008104 phosphatidylethanolamines Chemical class 0.000 description 1
- 150000003905 phosphatidylinositols Chemical class 0.000 description 1
- PTMHPRAIXMAOOB-UHFFFAOYSA-L phosphoramidate Chemical compound NP([O-])([O-])=O PTMHPRAIXMAOOB-UHFFFAOYSA-L 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 239000013573 pollen allergen Substances 0.000 description 1
- 230000008488 polyadenylation Effects 0.000 description 1
- 229920000056 polyoxyethylene ether Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 101150012212 prlC gene Proteins 0.000 description 1
- 238000011165 process development Methods 0.000 description 1
- 229940002612 prodrug Drugs 0.000 description 1
- 239000000651 prodrug Substances 0.000 description 1
- 125000001500 prolyl group Chemical group [H]N1C([H])(C(=O)[*])C([H])([H])C([H])([H])C1([H])[H] 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 108010064037 prorennin Proteins 0.000 description 1
- 108010079891 prostein Proteins 0.000 description 1
- 230000004952 protein activity Effects 0.000 description 1
- 238000003498 protein array Methods 0.000 description 1
- 229960000856 protein c Drugs 0.000 description 1
- 238000000751 protein extraction Methods 0.000 description 1
- 229940121649 protein inhibitor Drugs 0.000 description 1
- 239000012268 protein inhibitor Substances 0.000 description 1
- 108060006633 protein kinase Proteins 0.000 description 1
- 230000009145 protein modification Effects 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 210000001938 protoplast Anatomy 0.000 description 1
- 101150060364 ptrA gene Proteins 0.000 description 1
- 101150014928 ptrB gene Proteins 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 239000012521 purified sample Substances 0.000 description 1
- 150000003235 pyrrolidines Chemical class 0.000 description 1
- 229960003876 ranibizumab Drugs 0.000 description 1
- 230000007115 recruitment Effects 0.000 description 1
- 239000012925 reference material Substances 0.000 description 1
- 230000014493 regulation of gene expression Effects 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 239000002342 ribonucleoside Substances 0.000 description 1
- 238000007363 ring formation reaction Methods 0.000 description 1
- 210000003296 saliva Anatomy 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 108010073863 saruplase Proteins 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 229930195734 saturated hydrocarbon Natural products 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 150000003354 serine derivatives Chemical class 0.000 description 1
- 239000013605 shuttle vector Substances 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 229940126586 small molecule drug Drugs 0.000 description 1
- NRHMKIHPTBHXPF-TUJRSCDTSA-M sodium cholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 NRHMKIHPTBHXPF-TUJRSCDTSA-M 0.000 description 1
- FHHPUSMSKHSNKW-SMOYURAASA-M sodium deoxycholate Chemical compound [Na+].C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 FHHPUSMSKHSNKW-SMOYURAASA-M 0.000 description 1
- VGTPCRGMBIAPIM-UHFFFAOYSA-M sodium thiocyanate Chemical compound [Na+].[S-]C#N VGTPCRGMBIAPIM-UHFFFAOYSA-M 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000003270 steroid hormone Substances 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 125000003107 substituted aryl group Chemical group 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 125000002653 sulfanylmethyl group Chemical group [H]SC([H])([H])[*] 0.000 description 1
- 229940117986 sulfobetaine Drugs 0.000 description 1
- 125000000472 sulfonyl group Chemical group *S(*)(=O)=O 0.000 description 1
- IMCGHZIGRANKHV-AJNGGQMLSA-N tert-butyl (3s,5s)-2-oxo-5-[(2s,4s)-5-oxo-4-propan-2-yloxolan-2-yl]-3-propan-2-ylpyrrolidine-1-carboxylate Chemical compound O1C(=O)[C@H](C(C)C)C[C@H]1[C@H]1N(C(=O)OC(C)(C)C)C(=O)[C@H](C(C)C)C1 IMCGHZIGRANKHV-AJNGGQMLSA-N 0.000 description 1
- 229940124597 therapeutic agent Drugs 0.000 description 1
- 150000003573 thiols Chemical class 0.000 description 1
- 238000006257 total synthesis reaction Methods 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- IEDVJHCEMCRBQM-UHFFFAOYSA-N trimethoprim Chemical compound COC1=C(OC)C(OC)=CC(CC=2C(=NC(N)=NC=2)N)=C1 IEDVJHCEMCRBQM-UHFFFAOYSA-N 0.000 description 1
- 229960001082 trimethoprim Drugs 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 239000002753 trypsin inhibitor Substances 0.000 description 1
- ORHBXUUXSCNDEV-UHFFFAOYSA-N umbelliferone Chemical compound C1=CC(=O)OC2=CC(O)=CC=C21 ORHBXUUXSCNDEV-UHFFFAOYSA-N 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 229930195735 unsaturated hydrocarbon Natural products 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 238000011179 visual inspection Methods 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
- JLYXXMFPNIAWKQ-UHFFFAOYSA-N γ Benzene hexachloride Chemical compound ClC1C(Cl)C(Cl)C(Cl)C(Cl)C1Cl JLYXXMFPNIAWKQ-UHFFFAOYSA-N 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
- 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
- C12N15/70—Vectors or expression systems specially adapted for E. coli
-
- 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
- C07K14/24—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- C07K14/245—Escherichia (G)
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
- C07K16/2809—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against the T-cell receptor (TcR)-CD3 complex
-
- 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
-
- 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
- C12N15/67—General methods for enhancing the expression
-
- 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
- C12N9/90—Isomerases (5.)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y502/00—Cis-trans-isomerases (5.2)
- C12Y502/01—Cis-trans-Isomerases (5.2.1)
- C12Y502/01008—Peptidylprolyl isomerase (5.2.1.8), i.e. cyclophilin
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/52—Constant or Fc region; Isotype
- C07K2317/522—CH1 domain
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/55—Fab or Fab'
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Gastroenterology & Hepatology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
本出願は、2018年5月1日に出願された「A Method for Optimizing Antibody Expression」という名称の米国仮出願第62/665,093号の利益を主張するものであり、その内容は、その全体が参照により本明細書に組み込まれる。
本出願は、EFS−Webを介してASCIIフォーマットで提出され、その全体が参照により本明細書に組み込まれる配列表を含む。2019年4月30日に作成されたASCIIコピーの名称は、_AMBX_0226_00PCT_ST25.txtであり、サイズは15,184バイトである。
本開示は、分子生物学の分野に関する。本発明の開示は、ベクターおよびプラスミドの設計および操作に関する。本発明はまた、天然にコードされていないアミノ酸を組み込んだ新規な産生株または細胞にも関し、およびそれらから組み換えタンパク質を産生する方法にも関する。
大腸菌(E. coli)ペリプラズムにおける、適切に組み立てられた無傷の活性抗体フラグメントの蓄積が正確に発生するためには、以下を含む複数の段階が必要である:転写、翻訳、非天然アミノ酸取り込みのための終止コドンの抑制、内膜を横断する移動、シグナルペプチドの切断、適切な折り畳み、プロリン残基の正確なシスまたはトランス異性化、重鎖との軽鎖の組み立て、ジスルフィド内結合およびジスルフィド間結合の正確な形成、およびペリプラズムにおける保持(外膜を越えて培地への漏出がない)。細胞質またはペリプラズムにおけるポリペプチドの代替経路は、切断、誤った折り畳み、凝集、およびタンパク質分解を含む(Miot M. & Betton J.M., Microbial Cell Factories, 3, 4, 2004; Yoon, S. H. et al., Recent Patents in Biotech, 4, 23, 2010)。
本発明は、新規の操作されたベクター、プラスミドおよび産生株を提供する。いくつかの実施形態において、本発明は、タンパク質発現最適化および力価収率のための新規の発現プラスミドおよび対応する産生株を提供する。本発明の実施形態において、操作されたベクターおよび株は、部位特異的に組み込まれた非天然アミノ酸を有する目的のタンパク質をコードする核酸配列と、1つ以上の成分をコードする核酸配列とを含む。ここで、当該成分は、ヘルパータンパク質もしくは因子または遺伝要素(genetic element)である。他の実施形態において、本発明の開示は、大腸菌に限定されない原核生物系におけるタンパク質発現を最適化するための方法、ならびに発現プラスミドおよび産生株を提供する。
置換セリン等の糖置換アミノ酸;炭素結合糖含有アミノ酸;酸化還元活性アミノ酸;α−ヒドロキシ含有酸;アミノチオ酸含有アミノ酸;α,α−二置換アミノ酸;β−アミノ酸;およびプロリン以外の環状アミノ酸からなる群から選択される。
本発明の新規な特徴は、添付の特許請求の範囲に詳細に記載される。本発明の原理が利用される例示的な実施形態が記載されている以下の詳細な説明、および添付の図面を参照することによって、本発明の開示の特徴および利点のより良い理解が得られるであろう。
本発明を詳細に説明する前に、本発明は、特定の方法論、または組成物、または生物学的系に限定されず、これらは当然、変化し得ることが理解されるべきである。また、本明細書中で使用される用語は、特定の実施形態を説明することのみを目的としており、限定することを意図していないことを理解されたい。本明細書中および添付の特許請求の範囲中で使用されるとおり、単数形「a」、「an」、および「the」は、内容が別段の明確な指示をしない限り、複数の対象を含む。したがって、例えば、「細胞(a cell)」への言及は、2つ以上の細胞の組合せ等を含む。
用語「アミノ酸」は、天然に存在するアミノ酸、および非天然(non-natural)アミノ酸または非天然(unnatural)アミノ酸、ならびに天然に存在するアミノ酸と同様の様式で機能するアミノ酸アナログおよびアミノ酸模倣物をいう。天然にコードされたアミノ酸は、20個の共通アミノ酸(アラニン、アルギニン、アスパラギン、アスパラギン酸、システイン、グルタミン、グルタミン酸、グリシン、ヒスチジン、イソロイシン、ロイシン、リジン、メチオニン、フェニルアラニン、プロリン、セリン、スレオニン、トリプトファン、チロシン、およびバリン)、ピロリジン、およびセレノシステインである。アミノ酸アナログは、天然に存在するアミノ酸と同じ基本化学構造、ほんの一例として、水素、カルボキシル基、アミノ基、および官能R基に結合したα−炭素を有する化合物をいう。そのようなアナログは、修飾されたR基(例として、ノルロイシン)を有し得、または天然に存在するアミノ酸と同じ基本化学構造をなお保持しつつ修飾されたペプチド骨格を有し得る。アミノ酸アナログの非限定的な例は、ホモセリン、ノルロイシン、メチオニンスルホキシド、メチオニンメチルスルホニウムを含む。アミノ酸は、それらの名称、それらの一般に知られている3文字記号、またはIUPAC−IUB Biochemical Nomenclature Commissionによって推奨される1文字記号のいずれかによって、本明細書中で言及され得る。さらに、ヌクレオチドは、それらの一般に受け入れられている一文字コードによって、言及されてもよい。
本明細書中で使用される場合、「遺伝要素(genetic elements)」または「遺伝要素(genetic element)」は、タンパク質発現を促進するために、ヘルパータンパク質またはヘルパー因子と共に促進、増進、増強または相乗作用を与え得る任意の分子を含み得る。遺伝要素は、タンパク質発現に関連するまたは関与する1つ以上の遺伝子の転写を促進し得る。そのような要素は、非真核生物源または真核生物源由来であり得る。
原核生物系(例えば大腸菌)におけるタンパク質発現の最適化において、本発明は、天然にコードされていないアミノ酸が組み込まれている発現困難な(DTE)抗体断片を含むタンパク質を利用する。大腸菌における非天然アミノ酸取り込み技術の使用は、当技術分野で周知である。この技術は、原核生物種において機能する直交tRNA/アミノアシルtRNA合成酵素ペアを用いて、セレクターコドンに応答して非天然アミノ酸を部位特異的に組み込む。例えば、WO2002/085923、WO2002/086075、WO2004/09459、WO2005/019415、WO2005/007870およびWO2005/007624を参照されたい。これらの文献は、その内容全体が参照により組み込まれる。Wang and Schultz, "Expanding the Genetic Code," Angewandte Chemie Int. Ed., 44(1):34-66, 2005(その内容は、その全体が参照により組み込まれる)も参照されたい。非天然アミノ酸のタンパク質への組み込みは、非天然アミノ酸の組み込みをシグナル伝達するセレクターコドンを含むように目的のタンパク質(または遺伝子)をコードするポリヌクレオチドを操作することによって、任意の所望の位置で生じるようにプログラムされ得る。本発明のセレクターコドンは例えば、ユニークな3塩基コドン、ナンセンスコドン(例えば、終止コドン)を含む。
本開示は、その中に部位特異的に組み込まれた非天然に(non-naturally)または非天然に(unnaturally)コードされたアミノ酸を有する抗体断片を含む。抗体または抗体断片中に非天然アミノ酸を部位特異的に導入するための方法が当技術分野に記載されており、例えば、WO2010/011735およびWO2005/074650を参照のこと。典型的には、本発明の非天然アミノ酸は、20個の天然アミノ酸において利用できない追加の特徴を提供するように選択または設計される。例えば、非天然アミノ酸は、それらが組み込まれるタンパク質の生物学的特性を改変するように任意に設計または選択される。例えば、以下の特性は、タンパク質中に非天然アミノ酸を含めることによって任意に改変される:毒性、生体内分布、溶解度、安定性(例えば、耐熱性、耐加水分解性、酸化耐性、酵素分解に対する耐性等)、精製および加工の容易さ、構造特性、分光特性、化学的および/または光化学的特性、触媒活性、酸化還元電位、半減期、(共有結合または非共有結合のいずれかで)他の分子と反応する能力等。
クローン化ポリヌクレオチドの高レベル発現を得るために、通常は、転写を指示する強力なプロモーター、転写/翻訳ターミネーター、およびタンパク質をコードする核酸については翻訳開始のためのリボソーム結合部位を含む発現ベクターに、目的のタンパク質またはポリペプチドをコードするポリヌクレオチドをサブクローニングする。適切な細菌プロモーターは当業者に公知であり、そして例えば、Sambrook et al.およびAusubel et al.に記載される。
本開示は、当技術分野で周知の方法論およびルーチン技術を包含する。これらには、分子生物学、組換えDNA技術、生化学および細胞生物学の従来の方法が含まれ、これらは全て当技術分野の技術の範囲内である。そのような技術は、例えば、Molecular Cloning: A Laboratory Manual, Third Edition (Sambrook et al., Eds., Cold Spring Harbor Press, Cold Spring Harbor, NY, 2001);Oligonucleotide Synthesis: Methods And Applications (Methods in Molecular Biology), (Herdewijn, P., Ed., Humana Press, Totowa, NJ);Oligonucleotide Synthesis (Gait, M. J., Ed., 1984);Methods In Molecular Biology, (Humana Press, Totowa, NJ);Cell Biology: A Laboratory Notebook (Cellis, J. E., Ed., Academic Press, New York, NY, 1998);Animal Cell Culture (Freshney, R. I., Ed., 1987); Introduction To Cell And Tissue Culture (Mather, J. P. and Roberts, P. E., Eds., Plenum Press, New York, NY, 1998);Cell And Tissue Culture: Laboratory Procedures (Doyle, A. et al., Eds., John Wiley and Sons, Hoboken, NJ, 1993-8);Gene Transfer Vectors For Mammalian Cells (Miller, J. M. et al. Eds., Cold Spring Harbor Press, Cold Spring Harbor, NY, 1987);Current Protocols In Molecular Biology (Ausubel, F. M. et al., Eds., Greene Pub. Associates, New York, NY, 1987);PCR: The Polymerase Chain Reaction, (Mullis, K. et al., Eds., Birkhauser, Boston, MA, 1994);Short Protocols In Molecular Biology (John Wiley and Sons, Hoboken, NJ, 1999);Immunobiology 7 (Janeway, C. A. et al., Garland Science, London, UK, 2007);Antibodies (P. Finch, Stride Publications, Devoran, UK, 1997);Antibodies: A Practical Approach (D. Catty., ed., Oxford University Press, USA, New York, NY, 1989);Monoclonal Antibodies: A Practical Approach (Shepherd, P. et al. Eds., Oxford University Press, USA, New York NY, 2000);Using Antibodies: A Laboratory Manual (Harlow, E. et al. Eds., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1998);The Antibodies (Zanetti, M. et al. Eds., Harwood Academic Publishers, London, UK, 1995)等の文献において十分に説明されている。
本明細書中に記載される組成物のいずれも、適切な容器手段において、キット中で一緒に組み立てられ得る。より詳細には、本発明のベクターおよび/またはプラスミドを設計および構築するまたは操作するための構成要素の全てまたはサブセットが、キット中に一緒にパッケージングされ得る。本発明の1つまたは複数の構成要素は、別々にまたは一緒にパッケージングされ得る。いくつかの実施形態において、ベクターおよび/またはプラスミドを、本明細書中に開示される1つ以上の構成要素と一緒にパッケージングすることが好ましい。本発明の実施形態において、タンパク質、ペプチド、ポリペプチド(抗体断片を含む)、またはそれらをコードする核酸は、一緒に、または単一分子として、または分子のセットとしてパッケージングされ得る。また、本明細書中に開示される構成要素は、印刷物または機械で読み取り可能な媒体における説明書と共に個々にパッケージされ、そして販売され得、当該説明書は、本明細書中に開示されるように、ベクターおよび/またはプラスミドを設計および構築するために、それらが互いに関連してどのように使用され得るかを記載している。
本明細書中に記述された実施例および実施形態は、説明のみを目的とするものであり、それらに関する様々な改変または変更が当業者に示唆され、そして本出願の精神および範囲、ならびに添付された特許請求の範囲の内に含まれるべきであると理解される。
本発明の抗CD3 Fab発現プラスミドを、Gibson アセンブリキット(New England Biolabs、MA)を使用する組換えに基づくクローニング法によって、または以下に記載されるとおり、大腸菌NEB5αクローニング株(New England Biolabs、MA)においてQuikChange変異誘発キット(Agilent Technologies、CA)を使用することによって操作した。表1は、本研究で使用した3つの大腸菌クローニング宿主および産生宿主、そしてそれらの詳細な遺伝子型を示している。
本発明の産生株を調製するために、化学的にコンピテントな大腸菌W3110B60宿主細胞を、配列が確認されたプラスミドDNA(50ng)で形質転換し、組換え細胞を、50μg/mLの硫酸カナマイシン(Sigma)(またはカルベニシリン100μg/mL、Teknova)を含有する2×YT+1%グルコース寒天プレート上で選択し、そして37℃で一晩インキュベートした。次いで、新鮮な形質転換プレートからの単一コロニーを、50μg/mLの硫酸カナマイシンまたはカルベニシリン100μg/mLを含有する2×YT+1%グルコース寒天プレート上で、連続する三重画線(triple-streaking)および37℃で一晩インキュベートすることによって、3回増殖させた。最後に、第3回目の画線プレートからの単一コロニーを、50μg/mL硫酸カナマイシン(Sigma)(またはカルベニシリン100μg/mL、Teknova)を含有する20mL Super Broth(Fisher−OptiglowTM)に播種し、そして37℃および250rpmで一晩インキュベートした。次いで、一晩生育させた培養物を、最終グリセロール濃度を20%(w/v)になるようにグリセロールで希釈した。次いで、この細胞懸濁液を、いくつかのクライオバイアル中に1mlアリコートに分注し、そして産生株バイアルとして−80℃で凍結させた。
抗CD3 Fab−pAcFの産生のための発酵プロセスは、以下の2つのステージからなる:(i)播種材料調製および(ii)発酵槽産生。播種材料を単一のグリセロールバイアルから開始し、解凍し、250mLバッフル付き三角フラスコ中の50mLの規定種培地中に1:1000(v/v)に希釈し、そして37℃および250rpmでインキュベートする。使用前に、発酵槽を清潔にし、オートクレーブ処理する。特定量の基礎培地を発酵槽に添加し、蒸気滅菌する。特定量の硫酸カナマイシン溶液、フィード培地およびP2000消泡剤を、播種前に基礎培地に添加する。オートクレーブ処理後に発酵槽に添加した全ての溶液は、0.2μmフィルター濾過するか、または無菌添加の前にオートクレーブ処理する。産生発酵槽は、播種材料の内容物を無菌的に移すことによって、0.0004の標的OD600で播種される。播種後、OD600を決定するために適切なインターバルで培養物をサンプリングする。温度、pHおよび溶存酸素をモニターし、それぞれ37℃、7.0および≧30%の特定の設定点で制御する。pHは、水酸化アンモニウム溶液または硫酸の添加によって制御する。溶存酸素は、撹拌速度を変化させることによって、およびスパージした(sparged)空気/酸素混合物中の酸素の組成を増加させることによって制御する。発酵プロセスの間に消泡剤を添加して、発泡を制御する。
発酵試料のプラスミド保持率を測定するために、グリセロール(20%v/v)を、収穫時に1mL発酵培養液に添加し、そして将来の分析のために−80℃で保存した。この試料から100μLを採取し、そしてLB培養液900μLに、10−1希釈、10−2希釈、10−3希釈、10−4希釈、10−5希釈、10−6希釈、10−7希釈および10−8希釈によって連続希釈した。100μLの10−4希釈、10−5希釈、10−6希釈、10−7希釈および10−8希釈を、2×YT+1%グルコース寒天プレート上にプレートした。プレートを30℃で一晩インキュベートした。翌日、100個のコロニーを、ウェル単離されたコロニーを有する希釈プレートから拾い、そしてレプリカを、2×YT+1%グルコース寒天および2×YT+1%グルコース+カナマイシン(50μg/mL)プレート上にプレーティングした。プレートを30℃で一晩インキュベートした。一晩インキュベートした後、コロニーを計数し、そして2×YT+1%グルコースプレートと2×YT+1%グルコース+カナマイシン(50μg/mL)プレートとの間で比較した。プラスミド保持率は、2×YT+1%グルコース+カナマイシン(50μg/mL)プレートと比較した、2×YT+1%グルコースプレート上で増殖したコロニーのパーセンテージとして報告される。
産物力価を定量するために、1mLの発酵培養液を、発酵プロセス全体の様々な時点で遠心分離した(18,000×g、5分間)。上清を別の1.5mL遠心管に移し、ペレットおよび上清の両方を保持し、そして将来の分析のために−80℃で保存した。細胞ペレットを溶解し、そして以下に記載されるとおりELISAまたはCEXアッセイを使用して力価を決定した。
本発明の抗CD3 Fabを、大腸菌細胞中でそれぞれ産生させ、全細胞溶解物(WCL)上清から回収した。細胞溶解を4℃で行った。細胞を100mM酢酸、100mM NaCl、1mM EDTA(pH3.5)中に、もとの発酵体積に等しい体積で溶解し、溶解後pH4.1〜4.2となる。溶解後、WCLを15,900×gで30分間、4℃で遠心分離し、そして濾過して(0.8/0.2ミクロン)、沈殿したタンパク質および細胞の残骸を除去する。Capto Sカチオン交換クロマトグラフィーを使用して、大腸菌WCL上清から抗CD3 Fabを捕捉した。RP−HPLC分析をCapto Sプールで行って、さらに切り取られ且つ切断された重鎖および軽鎖種に対する無傷のFabの純度を決定した。前記さらに切り取られ且つ切断された重鎖および軽鎖種は、前記無傷のFabと共にCapto Sカラムで共精製される。回復可能力価はまた、Capto S捕捉カラムから回収されたタンパク質の総量を計算し、それにCapto S溶出プール中の無傷のFabのパーセンテージを乗じることによって推定した。溶解および精製のために収穫した細胞ペレットを再び懸濁して発酵収穫体積に戻したので、無傷のFabの量を初期Capto Sにロードした体積で割って、発酵力価を推定した。
図1は、示した重鎖定常ドメイン(CH1)内のUAA取り込み部位の位置を有する抗CD3 Fab発現カセットの模式図を示す。このバイシストロニックオペロンは大腸菌のアルカリホスファターゼ(phoA)プロモーターによって駆動され、培地中のリン酸塩が枯渇すると誘導される。抗CD3 Fabのアミノ酸配列は、vHおよびvL(それぞれ免疫グロブリン重鎖および軽鎖の可変領域)コード配列+ヒトγ1(CH1)およびκ(CL)定常領域から構成される。重鎖および軽鎖の両方に付加された分泌シグナルペプチドSTIIは、適切な折り畳みおよびジスルフィド結合形成のために、酸化ペリプラズムへのFabの分泌を指示する。アンバー終止コドン(TAG)を、抗CD3 Fab中の重鎖定常ドメイン(CH1)の129位におけるリジン(K)残基129で操作した(以下、HK129アンバーまたはHK129amと呼ぶ)(図1に示す)。このTAG終止コドンは、進化したM.ヤンナスキイ(M. jannaschii)のpAcF tyrRS/tRNA機構を用いてこの部位でpAcFをコードした(例えば、Wang L. et al., Science, 292, 498, 2001; Zhang Z. et. al., Biochemistry, 42, 6735, 2003を参照のこと)。
図3は、発現プラスミドp56のマップを示す。このプラスミドの設計および操作において、プラスミド内に含まれるE9 RS合成酵素およびtRNAクラスターを有する直交系を利用した(例えば、米国特許公開第2003010885号、同第20050009049号、同第20050208536号、同第20060233744号を参照のこと)。また、本発明のプラスミドの設計および操作において利用したのは、プロリン誤取り込みを防止するとともに、独自のプラットフォーム産生宿主W3110B60株の染色体内の温度感受性proS変異を補完してプラスミド維持を確実にするための、大腸菌プロリン合成酵素、ProS、タンパク質の独自のプラスミドへの過剰発現であった(例えば、Javahishvili, T. et. al., ACS Chem. Biol., 9, 87, 2014に記載されている)。
産生ホスト細胞株W3110B60は、株2797を作製するために使用され、ゲノムのproSプロリン合成酵素遺伝子における変異を含む(W375R)。この変異は、34℃超でゲノムのProSタンパク質を不活性にし、従って、37℃で成長が阻害された温度感受性W3110B60細胞株を作製する。目的の抗CD3 Fab遺伝子(GOI)に加えて、2797株中のp56プラスミドはまた、野生型大腸菌ProSタンパク質の発現のための大腸菌proS遺伝子も含んでいる。これは成長を可能にし、GOIプラスミドを37℃で維持するための選択圧を提供する。しかし、産生温度が34℃より低い場合には、プラスミドの維持に有利ではないかもしれない。カナマイシン耐性のためのkanR遺伝子はまたp56プラスミド上に存在し(図3)、抗生物質のカナマイシンを培地に添加したときにプラスミド維持のための選択圧を与える。これらの構成要素の両方を用いても、発酵の間の種々の時点で収集した2797細胞において、乏しいプラスミド保持率が観察された(表4)。
前述のとおり、AfeI制限酵素サイトとNotI制限酵素サイトとの間において、parB遺伝子座を順方向(F)に挿入すると、文献から予想されるとおり、プラスミド保持率が改善された(100%)。しかし、予想外に、それはまた、細胞ペレット中で無傷のFab力価を約40%まで低下させた。この産生の損失を取り戻すために、プラスミド骨格配列を少しも欠失しない4種類の新しい発現プラスミド(p117、p118、p119およびp120;表2)を操作した。プラスミド骨格内のAfeIユニーク制限酵素サイトおよびZraIユニーク制限酵素サイトを利用した。同じparB遺伝子座を、Fab CDSに対して順方向(F)または逆方向(R)のいずれかの両方の位置にクローニングした。このストラテジーは、4つの新しい産生株、2884、2885、2886および2887(表2)を生じ、これらを、その後、プラスミド保持率および産生力価の両方について、元の株2797と高細胞密度発酵において比較した(図5A〜Bおよび表6)。
大腸菌におけるタンパク質力価の最適化または増加における分子シャペロンの関与を決定するために、ペプチジルプロリルイソメラーゼ活性を有する2つの周知の二機能性ペリプラズムシャペロン、FkpAおよびSkpを分析した。
上に開示したとおり、和合性を有する2プラスミド系においてFab力価の改善が観察されなかったことを考慮して、単一のプラスミド系を試験した。この系において、2886株における場合と同様、parB−ZraI−Fを有する抗CD3 Fabと同じプラスミド中に、シャペロン遺伝子FkpAおよびSkpを、天然プロモーターの後ろにクローニングした(図7B参照)。
株2909において上に開示したとおり、FkpAシャペロンを、単一プラスミド発現系において、parB遺伝子座の後ろに順方向にクローン化した(図10)。株2840における2プラスミド発現系のように別々のプラスミドから発現させた場合(図7A〜Bおよび図8を参照)、同じFkpAシャペロン遺伝子はFab力価を改善せず、これは、ヘルパーシャペロン発現を細胞生理と連携させることの重要性を強調するものである。
抗体Fabは、大腸菌において産生される場合、いかなる非天然アミノ酸の挿入もない産生条件下でペリプラズム中のプロテアーゼによって切り取られることが、文献において知られている(Battersby, J. E. et. al., Journal of Chromatography A, 927, 61, 2001)。Fab産生物の品質が直交アンバー抑制系によってどのように影響されるかを調べるために、無傷のFabならびにいくつかの追加の断片を捕捉するCapto S樹脂を使用するカチオン交換によって、Fab捕捉カラムを開発した。parB遺伝子座を有するプラスミドp119(表2)を保有する株2886由来のタンパク質を、本研究に利用した。
重鎖(HC)C末端伸長株の構築:前記の実施例に開示されるとおり、Capto S溶出プールにおけるRP−HPLCおよびインタクトマス(intact mass)は、全Fabタンパク質の内、高いパーセンテージで存在する種々のFab断片を同定した。Fab切断に関与する可能性のあるプロテアーゼは、尾部特異的プロテアーゼ(Tsp:tail-specific protease)としても知られる、ペリプラズムセリンエンドプロテアーゼPrc(C末端のプロセシングのための)である(Chen C. et al., Biotechnology and Bioengineering, 85, 463, 2004)。Prcは、タンパク質のC末端を認識し、そしてそれに結合し、次いで、タンパク質配列内の緩い領域または折り畳まれていない領域で切断すると考えられている(Keiler et al., Protein Science, 4, 1507, 1995)。切断サイトは、タンパク質の二次構造または三次構造によって決定され、一次配列によっては決定されず、従って、Prcは、タンパク質分解部位に関して広い配列特異性を有している。しかしながら、タンパク質のC末端配列は、Prcによるその切断に影響を及ぼし、そしてタンパク質のC末端配列を修飾することは、Prcによるそのタンパク質の切断量を変化させ得ることが示されている(Keiler K.C. & Sauer R.T., The Journal of Biological Chemistry, 271, 2589, 1996)。
本願において引用される全ての刊行物、特許、特許出願、および/または他の文献は、あたかも個々の刊行物、特許、特許出願、および/または他の文献が全ての目的のために参照により組み込まれることが個別に示されたかのように、同じ程度まで全ての目的のためにその全体が参照により組み込まれる。
Claims (39)
- 部位特異的に組み込まれた天然にコードされていないアミノ酸を有している目的のタンパク質をコードする核酸配列と、
1つ以上の成分をコードする核酸配列と、を含み、
前記1つ以上の成分は、配列番号12、15、または17から選択される、タンパク質発現を最適化するためのベクター。 - 前記1つ以上の成分は、ヘルパータンパク質もしくは因子または遺伝要素である、請求項1に記載のベクター。
- 前記ヘルパータンパク質または因子は、シャペロンである、請求項2に記載のベクター。
- 前記遺伝要素は、分配B遺伝子座(parB)である、請求項2に記載のベクター。
- 抗体重鎖C末端末伸長変異体をさらに含む、請求項1に記載のベクター。
- 前記重鎖C末端末伸長変異体は、1つ以上のアミノ酸を含む、請求項5に記載のベクター。
- parBは、前記目的のタンパク質をコードする前記核酸配列のAfe1サイトとNot1サイトとの間において、順転写方向に位置する、請求項1に記載のベクター。
- parBは、前記目的のタンパク質をコードする前記核酸配列のAfe1サイトまたはZra1サイトにおいて、順転写方向に位置する、請求項1に記載のベクター。
- parBは、前記目的のタンパク質をコードする前記核酸配列のAfe1サイトまたはZra1サイトにおいて、逆転写方向に位置する、請求項1に記載のベクター。
- 前記シャペロンは、SkpまたはFkpAである、請求項3に記載のベクター。
- 前記シャペロンは、SkpおよびFkpAである、請求項3に記載のベクター。
- 分配B遺伝子座(parB)をさらに含む、請求項10または11に記載のベクター。
- 前記目的のタンパク質は、バイオセラピューティック、免疫原、抗体、抗体断片またはそれらの変異体から選択される、請求項1に記載のベクター。
- 前記バイオセラピューティックは、ワクチンである、請求項13に記載のベクター。
- 前記目的のタンパク質は、サイトカイン、ケモカイン、成長因子、成長因子受容体、インターフェロン、インターロイキン、炎症性分子、がん遺伝子産物、ペプチドホルモン、シグナル伝達分子、またはステロイドホルモン受容体である、請求項13に記載のベクター。
- 前記目的のタンパク質は、HER2、CD−70、PSMA、5T4、EGFR、TROP2、CD3、IL−2、IL−3、IL−10、IL−12、IL−15、IL−21、GPC3、DLL3、ROR1、レプチン、グレリン、FGF−1、FGF−19、FGF−21、FGF−23、HGH、FcR、インスリン、IGF1、TNFR1、TRAIL、EPO、およびそれらのアナログ、二重特異性体または断片である、請求項13に記載のベクター。
- 前記抗体断片は、Fab、Fab’、F(ab’)2、Fv断片、一本鎖抗体断片(scFv)、ジスルフィド安定化scFv(dsFv))、ダイアボディ(Db)、BiTE(二重特異性T細胞エンゲージャー)、DART(二重親和性再ターゲティング)、またはタンデムダイアボディ(TandAb)である、請求項13に記載のベクター。
- 前記抗体断片は、Fabである、請求項17に記載のベクター。
- 前記抗体断片は、抗CD3 Fabである、請求項13に記載のベクター。
- 部位特異的に組み込まれた天然にコードされていないアミノ酸を有している目的のタンパク質をコードする核酸配列と、
1つ以上の成分をコードする核酸配列と、を含み、
前記1つ以上の成分は、配列番号12、15、または17から選択される、組換え細胞。 - 前記細胞は、株2797、株2835、株2884、株2885、株2886、株2887、株2909、株2910、株2914、株2915、株2916、株2917、株2918、株2839、株2840、株2841、株3004、株3005または株3006である、請求項20に記載の組換え細胞。
- 前記細胞は、シャペロンSkpおよび分配B遺伝子座(parB)を含む、請求項20に記載の組換え細胞。
- 前記細胞は、株2910または株2841から選択される、請求項22に記載の組換え細胞。
- 前記細胞は、シャペロンFkpAおよび分配B遺伝子座(parB)を含む、請求項20に記載の組換え細胞。
- 前記細胞は、株2909、株2840、株3004、株3005または株3006から選択される、請求項24に記載の組換え細胞。
- 前記細胞は、抗体重鎖C末端末伸長変異体を含む、請求項20に記載の組換え細胞。
- 前記細胞は、株2914、株2915、株2916、株2917、または株2918から選択される、請求項26に記載の組換え細胞。
- 前記プラスミドは、p56、p94、p96、p117、p118、p119、p120、p134、p135、p141、p142、p143、p144、p145、p199、p200またはp201である、先行する請求項のいずれかに記載の発現プラスミド。
- 天然にコードされていないアミノ酸を組み込んでいる目的の組換えタンパク質を産生する方法であって、
当該方法は、細菌細胞中で前記タンパク質を発現させる工程、および
1つ以上の成分をコードする核酸配列を導入する工程、を含み、
前記1つ以上の成分は、配列番号12、15、または17から選択される、方法。 - 部位特異的に組み込まれた非天然アミノ酸を有しているタンパク質の収量を改善するための、請求項29に記載の方法。
- 前記非天然アミノ酸は、O−メチル−L−チロシン、L−3−(2−ナフチル)アラニン、3−メチル−フェニルアラニン、O−4−アリル−L−チロシン、4−プロピル−L−チロシン、p−プロパルギルオキシ−L−フェニルアラニン、トリ−O−アセチル−GlcNAcβ−セリン、L−Dopa、フッ素化フェニルアラニン、イソプロピル−L−フェニルアラニン、p−アジド−L−フェニルアラニン、p−アシル−L−フェニルアラニン、p−ベンゾイル−L−フェニルアラニン、L−ホスホセリン、ホスホノセリン、ホスホノチロシン、p−ヨード−フェニルアラニン、p−ブロモフェニルアラニン、p−アミノ−L−フェニルアラニン、イソプロピル−L−フェニルアラニン、パラ−アセチルフェニルアラニン、p−ニトロフェニルアラニン、p−スルホチロシン、p−カルボキシフェニルアラニン、o−ニトロフェニルアラニン、m−ニトロフェニルアラニン、p−ボロニルフェニルアラニン、o−ボロニルフェニルアラニン、m−ボロニルフェニルアラニン、p−アミノフェニルアラニン、o−アミノフェニルアラニン、m−アミノフェニルアラニン、p−アシルフェニルアラニン、o−アシルフェニルアラニン、m−アシルフェニルアラニン、p−OMeフェニルアラニン、o−OMeフェニルアラニン、m−OMeフェニルアラニン、p−スルホフェニルアラニン、o−スルホフェニルアラニン、m−スルホフェニルアラニン、5−ニトロHis、3−ニトロTyr、2−ニトロTyr、ニトロ置換Leu、ニトロ置換His、ニトロ置換De、ニトロ置換Trp、2−ニトロTrp、4−ニトロTrp、5−ニトロTrp、6−ニトロTrp、7−ニトロTrp、3−アミノチロシン、2−アミノチロシン、O−スルホチロシン、2−スルホオキシフェニルアラニン、3−スルホオキシフェニルアラニン、o−カルボキシフェニルアラニン、m−カルボキシフェニルアラニン、p−アセチル−L−フェニルアラニン、p−プロパルギル−フェニルアラニン、O−メチル−L−チロシン、L−3−(2−ナフチル)アラニン、3−メチル−フェニルアラニン、O−4−アリル−L−チロシン、4−プロピル−L−チロシン、トリ−O−アセチル−GlcNAcβ−セリン、L−Dopa、フッ素化フェニルアラニン、イソプロピル−L−フェニルアラニン、p−アジド−L−フェニルアラニン、p−アシル−L−フェニルアラニン、p−ベンゾイル−L−フェニルアラニン、L−ホスホセリン、ホスホノセリン、ホスホノチロシン、p−ヨード−フェニルアラニン、p−ブロモフェニルアラニン、p−アミノ−L−フェニルアラニン、イソプロピル−L−フェニルアラニンまたはp−プロパルギルオキシ−フェニルアラニンである、請求項30に記載の方法。
- 前記非天然アミノ酸は、パラ−アセチルフェニルアラニンである、請求項31に記載の方法。
- 請求項29に記載の単離された細胞または細胞株。
- 非天然アミノ酸含有タンパク質またはポリペプチドを産生するための、請求項33に記載の単離された細胞または細胞株。
- 前記非天然アミノ酸含有タンパク質の前記収量は、1つ以上のシャペロンおよびparB遺伝子座の存在下で少なくとも0.25倍以上多い、請求項29に記載の単離された細胞または細胞株。
- 前記parB遺伝子座は、前記目的のタンパク質をコードする前記核酸配列のAfe1サイトにおいて、逆転写方向に位置する、請求項35に記載の単離された細胞または細胞株。
- 産生細胞または細胞株におけるプラスミド保持率を最適化するための、請求項35に記載の単離された細胞または細胞株。
- 前記抗体は、完全長抗体である、請求項17に記載のベクター。
- 前記完全長抗体は、IgG1抗体、IgG2抗体、IgG3抗体、またはIgG4抗体である、請求項38に記載のベクター。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201862665093P | 2018-05-01 | 2018-05-01 | |
US62/665,093 | 2018-05-01 | ||
PCT/US2019/030295 WO2019213331A1 (en) | 2018-05-01 | 2019-05-01 | A method for optimizing antibody expression |
Publications (1)
Publication Number | Publication Date |
---|---|
JP2021522785A true JP2021522785A (ja) | 2021-09-02 |
Family
ID=66776873
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2020560954A Pending JP2021522785A (ja) | 2018-05-01 | 2019-05-01 | 抗体発現を最適化するための方法 |
Country Status (11)
Country | Link |
---|---|
US (1) | US20210108213A1 (ja) |
EP (1) | EP3788149A1 (ja) |
JP (1) | JP2021522785A (ja) |
KR (1) | KR20210005172A (ja) |
CN (1) | CN112313336A (ja) |
AU (1) | AU2019263303A1 (ja) |
BR (1) | BR112020022288A2 (ja) |
CA (1) | CA3098910A1 (ja) |
MX (1) | MX2020011529A (ja) |
SG (1) | SG11202010439VA (ja) |
WO (1) | WO2019213331A1 (ja) |
Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20230031981A (ko) | 2019-05-14 | 2023-03-07 | 프로벤션 바이오, 인코포레이티드 | 제1형 당뇨병을 예방하기 위한 방법 및 조성물 |
CN111304234A (zh) * | 2020-02-27 | 2020-06-19 | 江南大学 | 一种适用于枯草芽孢杆菌的非天然氨基酸利用工具 |
AU2021287998A1 (en) | 2020-06-11 | 2023-02-02 | Benaroya Research Institute At Virginia Mason | Methods and compositions for preventing type 1 diabetes |
EP4223778A1 (en) * | 2020-09-29 | 2023-08-09 | Innovent Biologics (Suzhou) Co., Ltd. | Anti-claudin18.2 and cd3 bispecific antibody and use thereof |
CN113699124B (zh) * | 2021-09-08 | 2022-04-12 | 北京大学 | 一种含非天然氨基酸蛋白的制备方法 |
CN113862239B (zh) * | 2021-09-09 | 2023-06-30 | 山西中医药大学 | 一种核酸酶活性提高的蒙古黄芪病程相关蛋白突变体及其应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0759192B2 (ja) * | 1982-09-16 | 1995-06-28 | ベンツォン ファーマ エイ/エス | 不安定に遺伝されたプラスミドの安定化法 |
JP2014505482A (ja) * | 2011-02-03 | 2014-03-06 | ゾーマ テクノロジー リミテッド | 細菌中の機能的タンパク質発現を向上させるための方法および物質 |
JP2017501728A (ja) * | 2013-12-23 | 2017-01-19 | ザイムワークス インコーポレイティド | C末端軽鎖ポリペプチド伸長を含有する抗体、ならびにその複合体及びその使用方法 |
Family Cites Families (23)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4870008A (en) | 1983-08-12 | 1989-09-26 | Chiron Corporation | Secretory expression in eukaryotes |
HU206897B (en) | 1985-10-25 | 1993-01-28 | Zymogenetics Inc | Process for utilizing bar-1 gen for selecting strange proteins |
EP0742832A1 (en) * | 1994-02-03 | 1996-11-20 | Center For Innovative Technology | High expression nucleic acid vectors |
US5639635A (en) | 1994-11-03 | 1997-06-17 | Genentech, Inc. | Process for bacterial production of polypeptides |
US6083715A (en) | 1997-06-09 | 2000-07-04 | Board Of Regents, The University Of Texas System | Methods for producing heterologous disulfide bond-containing polypeptides in bacterial cells |
JP2000083670A (ja) | 1998-09-09 | 2000-03-28 | Hsp Kenkyusho:Kk | DsbA/DsbB/DsbC/DsbD発現プラスミド |
DK2322631T3 (en) | 2001-04-19 | 2015-01-12 | Scripps Research Inst | Methods and compositions for the production of orthogonal tRNA-aminoacyl-tRNA syntetasepar |
US6536736B2 (en) | 2001-07-16 | 2003-03-25 | Agilent Technologies, Inc. | Optomechanical mount for precisely steering/positioning a light beam |
US7429593B2 (en) | 2001-09-14 | 2008-09-30 | Shionogi & Co., Ltd. | Utilities of amide compounds |
AU2003250559A1 (en) | 2002-07-23 | 2004-02-09 | Yong-Seok Jeong | Beverage can |
KR20050073559A (ko) | 2002-10-16 | 2005-07-14 | 더 스크립스 리서치 인스티튜트 | 당단백질 합성 |
US7642085B2 (en) | 2002-12-22 | 2010-01-05 | The Scripps Research Institute | Protein arrays |
CN103289960B (zh) | 2003-04-17 | 2017-04-26 | 斯克利普斯研究院 | 扩展真核生物遗传密码 |
CA2531146A1 (en) | 2003-07-07 | 2005-03-03 | The Scripps Research Institute | Compositions of orthogonal lysyl-trna and aminoacyl-trna synthetase pairs and uses thereof |
WO2005007624A2 (en) | 2003-07-07 | 2005-01-27 | The Scripps Research Institute | Compositions of orthogonal glutamyl-trna and aminoacyl trna synthetase pairs and uses thereof |
WO2005007870A2 (en) | 2003-07-07 | 2005-01-27 | The Scripps Research Institute | COMPOSITIONS OF ORTHOGONAL LEUCYL-tRNA AND AMINOACYL-tRNA SYNTHETASE PAIRS AND USES THEREOF |
US8778880B2 (en) | 2004-02-02 | 2014-07-15 | Ambrx, Inc. | Human growth hormone modified at position 35 |
NZ551335A (en) * | 2004-06-18 | 2010-03-26 | Ambrx Inc | Antibodies and fragments thereof comprising an non naturally encoded amino acid coupled to a linker |
ES2529065T3 (es) * | 2005-12-14 | 2015-02-16 | Ambrx, Inc. | Composiciones que contienen, procedimientos que implican y usos de aminoácidos no naturales y polipéptidos |
EP2217702A4 (en) * | 2007-11-02 | 2012-05-16 | Scripps Research Inst | EVOLUTION DIRECTED USING PROTEINS COMPRISING NON-NATURAL AMINO ACIDS |
NZ600382A (en) | 2008-07-23 | 2013-11-29 | Ambrx Inc | Modified bovine G-CSF polypeptides and their uses |
GB201208367D0 (en) * | 2012-05-14 | 2012-06-27 | Ucb Pharma Sa | Biological product |
GB201419109D0 (en) * | 2014-10-27 | 2014-12-10 | Medical Res Council | Incorporation of unnatural amino acids into proteins |
-
2019
- 2019-05-01 SG SG11202010439VA patent/SG11202010439VA/en unknown
- 2019-05-01 BR BR112020022288-7A patent/BR112020022288A2/pt unknown
- 2019-05-01 MX MX2020011529A patent/MX2020011529A/es unknown
- 2019-05-01 WO PCT/US2019/030295 patent/WO2019213331A1/en unknown
- 2019-05-01 AU AU2019263303A patent/AU2019263303A1/en active Pending
- 2019-05-01 CA CA3098910A patent/CA3098910A1/en active Pending
- 2019-05-01 EP EP19729108.1A patent/EP3788149A1/en active Pending
- 2019-05-01 CN CN201980041409.9A patent/CN112313336A/zh active Pending
- 2019-05-01 JP JP2020560954A patent/JP2021522785A/ja active Pending
- 2019-05-01 KR KR1020207034038A patent/KR20210005172A/ko not_active Application Discontinuation
- 2019-05-01 US US17/051,690 patent/US20210108213A1/en active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPH0759192B2 (ja) * | 1982-09-16 | 1995-06-28 | ベンツォン ファーマ エイ/エス | 不安定に遺伝されたプラスミドの安定化法 |
JP2014505482A (ja) * | 2011-02-03 | 2014-03-06 | ゾーマ テクノロジー リミテッド | 細菌中の機能的タンパク質発現を向上させるための方法および物質 |
JP2017501728A (ja) * | 2013-12-23 | 2017-01-19 | ザイムワークス インコーポレイティド | C末端軽鎖ポリペプチド伸長を含有する抗体、ならびにその複合体及びその使用方法 |
Non-Patent Citations (4)
Title |
---|
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, vol. 38, JPN6023012849, 1992, pages 91 - 93, ISSN: 0005028201 * |
MICROBIAL CELL FACTORIES, vol. 9, no. 1, JPN6023012850, 2010, pages 22, ISSN: 0005028202 * |
PROC. NATL. ACAD. SCI. USA, vol. 83, JPN6023012852, 1986, pages 3116 - 3120, ISSN: 0005028204 * |
THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 275, no. 22, JPN6023012851, 2000, pages 17100 - 17105, ISSN: 0005028203 * |
Also Published As
Publication number | Publication date |
---|---|
BR112020022288A2 (pt) | 2021-02-23 |
EP3788149A1 (en) | 2021-03-10 |
KR20210005172A (ko) | 2021-01-13 |
SG11202010439VA (en) | 2020-11-27 |
CA3098910A1 (en) | 2019-11-07 |
US20210108213A1 (en) | 2021-04-15 |
AU2019263303A1 (en) | 2020-12-24 |
MX2020011529A (es) | 2021-01-29 |
WO2019213331A1 (en) | 2019-11-07 |
CN112313336A (zh) | 2021-02-02 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP2021522785A (ja) | 抗体発現を最適化するための方法 | |
US8198044B2 (en) | Systems for the expression of orthogonal translation components eubacterial host cells | |
AU2002256292C1 (en) | In vivo incorporation of unnatural amino acids | |
US9624485B2 (en) | Genetic incorporation of unnatural amino acids into proteins in mammalian cells | |
CN104328086A (zh) | 通过脊椎动物细胞位点特异性并入非天然氨基酸 | |
JP7245786B2 (ja) | 非天然アミノ酸含有タンパク質の産生を促進する方法および組成物 | |
EP3506945A1 (en) | Production of seleno-biologics in genomically recoded organisms | |
CN101541955A (zh) | 用于脊椎动物细胞的杂合抑制tRNA | |
WO2010114615A2 (en) | A facile system for encoding unnatural amino acids in mammalian cells | |
AU2008200780B2 (en) | In vivo incorporation of unnatural amino acids | |
KR20230022169A (ko) | 대장균을 이용한 전장 항체를 생산하는 방법 | |
JP2005046066A (ja) | 蛋白質生産用の新規なバクテリア及び蛋白質の製造方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20210112 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20220426 |
|
A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20230317 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20230404 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20230626 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20230904 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20231004 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20240109 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20240405 |