JP5935222B2 - タンパク質を精製するプロセス - Google Patents
タンパク質を精製するプロセス Download PDFInfo
- Publication number
- JP5935222B2 JP5935222B2 JP2013521208A JP2013521208A JP5935222B2 JP 5935222 B2 JP5935222 B2 JP 5935222B2 JP 2013521208 A JP2013521208 A JP 2013521208A JP 2013521208 A JP2013521208 A JP 2013521208A JP 5935222 B2 JP5935222 B2 JP 5935222B2
- Authority
- JP
- Japan
- Prior art keywords
- antibody
- recombinant
- protein
- seq
- host cell
- 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.)
- Active
Links
- 238000000034 method Methods 0.000 title claims description 101
- 230000008569 process Effects 0.000 title claims description 9
- 238000001742 protein purification Methods 0.000 title description 2
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 claims description 76
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 claims description 76
- 102000006010 Protein Disulfide-Isomerase Human genes 0.000 claims description 58
- 108020003519 protein disulfide isomerase Proteins 0.000 claims description 58
- 241000588724 Escherichia coli Species 0.000 claims description 53
- 239000000284 extract Substances 0.000 claims description 46
- 239000012634 fragment Substances 0.000 claims description 42
- 238000000746 purification Methods 0.000 claims description 23
- 238000010438 heat treatment Methods 0.000 claims description 15
- 230000001580 bacterial effect Effects 0.000 claims description 12
- 238000001914 filtration Methods 0.000 claims description 12
- 238000005119 centrifugation Methods 0.000 claims description 11
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 claims description 9
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 claims description 9
- 238000000926 separation method Methods 0.000 claims description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 8
- 238000004587 chromatography analysis Methods 0.000 claims description 8
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 6
- 230000001376 precipitating effect Effects 0.000 claims description 5
- 229960000583 acetic acid Drugs 0.000 claims description 4
- 239000011536 extraction buffer Substances 0.000 claims description 4
- 239000012362 glacial acetic acid Substances 0.000 claims description 4
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims description 4
- 101000611183 Homo sapiens Tumor necrosis factor Proteins 0.000 claims description 2
- 102000057041 human TNF Human genes 0.000 claims description 2
- 238000004255 ion exchange chromatography Methods 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 180
- 108090000623 proteins and genes Proteins 0.000 description 131
- 102000004169 proteins and genes Human genes 0.000 description 104
- 235000018102 proteins Nutrition 0.000 description 99
- 239000000523 sample Substances 0.000 description 62
- 238000010979 pH adjustment Methods 0.000 description 33
- 239000000243 solution Substances 0.000 description 33
- 229960003115 certolizumab pegol Drugs 0.000 description 27
- 230000014509 gene expression Effects 0.000 description 26
- 239000013612 plasmid Substances 0.000 description 26
- 235000001014 amino acid Nutrition 0.000 description 22
- 230000035772 mutation Effects 0.000 description 21
- 125000003275 alpha amino acid group Chemical group 0.000 description 20
- 239000012636 effector Substances 0.000 description 20
- 108020004414 DNA Proteins 0.000 description 19
- 150000001413 amino acids Chemical class 0.000 description 19
- 239000000427 antigen Substances 0.000 description 19
- 108091007433 antigens Proteins 0.000 description 19
- 102000036639 antigens Human genes 0.000 description 19
- 229920000642 polymer Polymers 0.000 description 19
- 230000002829 reductive effect Effects 0.000 description 19
- 101150081864 Spr gene Proteins 0.000 description 17
- 230000000694 effects Effects 0.000 description 17
- 101710196859 Dipeptide-binding protein Proteins 0.000 description 16
- 238000004458 analytical method Methods 0.000 description 16
- 230000001225 therapeutic effect Effects 0.000 description 15
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 13
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 13
- 108091005804 Peptidases Proteins 0.000 description 13
- 239000004365 Protease Substances 0.000 description 13
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 13
- -1 peptidoglycans Substances 0.000 description 13
- 238000004007 reversed phase HPLC Methods 0.000 description 13
- 108010076504 Protein Sorting Signals Proteins 0.000 description 12
- 229930006000 Sucrose Natural products 0.000 description 12
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 12
- 229960005091 chloramphenicol Drugs 0.000 description 12
- 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 12
- 239000013604 expression vector Substances 0.000 description 12
- 238000004519 manufacturing process Methods 0.000 description 12
- 239000002244 precipitate Substances 0.000 description 12
- 238000001556 precipitation Methods 0.000 description 12
- 239000005720 sucrose Substances 0.000 description 12
- 238000000605 extraction Methods 0.000 description 11
- 230000012010 growth Effects 0.000 description 11
- 239000003550 marker Substances 0.000 description 11
- 239000000047 product Substances 0.000 description 11
- 229920001817 Agar Polymers 0.000 description 10
- 239000008272 agar Substances 0.000 description 10
- 238000000855 fermentation Methods 0.000 description 10
- 230000004151 fermentation Effects 0.000 description 10
- 108091033319 polynucleotide Proteins 0.000 description 10
- 102000040430 polynucleotide Human genes 0.000 description 10
- 239000002157 polynucleotide Substances 0.000 description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 238000007792 addition Methods 0.000 description 9
- 239000000872 buffer Substances 0.000 description 9
- 229920001223 polyethylene glycol Polymers 0.000 description 9
- 108090000765 processed proteins & peptides Proteins 0.000 description 9
- 238000005516 engineering process Methods 0.000 description 8
- 239000000411 inducer Substances 0.000 description 8
- 210000001322 periplasm Anatomy 0.000 description 8
- 238000012545 processing Methods 0.000 description 8
- 239000000126 substance Substances 0.000 description 8
- 108091028043 Nucleic acid sequence Proteins 0.000 description 7
- 239000003795 chemical substances by application Substances 0.000 description 7
- 239000000356 contaminant Substances 0.000 description 7
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 7
- 230000001939 inductive effect Effects 0.000 description 7
- 229920001427 mPEG Polymers 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 239000002773 nucleotide Substances 0.000 description 7
- 125000003729 nucleotide group Chemical group 0.000 description 7
- 229920001184 polypeptide Polymers 0.000 description 7
- 102000004196 processed proteins & peptides Human genes 0.000 description 7
- 238000013518 transcription Methods 0.000 description 7
- 230000035897 transcription Effects 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 238000001042 affinity chromatography Methods 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 150000001875 compounds Chemical class 0.000 description 6
- 230000006698 induction Effects 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 230000004048 modification Effects 0.000 description 6
- 238000012986 modification Methods 0.000 description 6
- 230000010076 replication Effects 0.000 description 6
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 6
- 238000003860 storage Methods 0.000 description 6
- 108060003951 Immunoglobulin Proteins 0.000 description 5
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 5
- 108020004511 Recombinant DNA Proteins 0.000 description 5
- 238000004113 cell culture Methods 0.000 description 5
- 238000011210 chromatographic step Methods 0.000 description 5
- 230000002759 chromosomal effect Effects 0.000 description 5
- 230000007423 decrease Effects 0.000 description 5
- 238000004520 electroporation Methods 0.000 description 5
- 102000018358 immunoglobulin Human genes 0.000 description 5
- 230000010354 integration Effects 0.000 description 5
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 5
- 230000004952 protein activity Effects 0.000 description 5
- 239000003053 toxin Substances 0.000 description 5
- 231100000765 toxin Toxicity 0.000 description 5
- 108700012359 toxins Proteins 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 108010079246 OMPA outer membrane proteins Proteins 0.000 description 4
- 108091034117 Oligonucleotide Proteins 0.000 description 4
- 101710167893 Penicillin-binding protein 3 Proteins 0.000 description 4
- 101710146026 Peptidoglycan D,D-transpeptidase FtsI Proteins 0.000 description 4
- 239000007983 Tris buffer Substances 0.000 description 4
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 4
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 4
- 125000003277 amino group Chemical group 0.000 description 4
- 238000005277 cation exchange chromatography Methods 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 230000029087 digestion Effects 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 210000005256 gram-negative cell Anatomy 0.000 description 4
- 230000001976 improved effect Effects 0.000 description 4
- 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 4
- 239000008101 lactose Substances 0.000 description 4
- 125000005647 linker group Chemical group 0.000 description 4
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 101150036535 prc gene Proteins 0.000 description 4
- 230000028327 secretion Effects 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 229920001059 synthetic polymer Polymers 0.000 description 4
- 230000005945 translocation Effects 0.000 description 4
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 241001302584 Escherichia coli str. K-12 substr. W3110 Species 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical group OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 3
- 108010006785 Taq Polymerase Proteins 0.000 description 3
- 102220513284 Vasopressin V1b receptor_Y115F_mutation Human genes 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 210000004899 c-terminal region Anatomy 0.000 description 3
- 230000006037 cell lysis Effects 0.000 description 3
- 238000010367 cloning Methods 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 230000002950 deficient Effects 0.000 description 3
- 210000004408 hybridoma Anatomy 0.000 description 3
- 230000001965 increasing effect Effects 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 238000010369 molecular cloning Methods 0.000 description 3
- 229920005615 natural polymer Polymers 0.000 description 3
- 230000003204 osmotic effect Effects 0.000 description 3
- 239000008188 pellet Substances 0.000 description 3
- 230000012846 protein folding Effects 0.000 description 3
- 239000012460 protein solution Substances 0.000 description 3
- 102000005962 receptors Human genes 0.000 description 3
- 108020003175 receptors Proteins 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 102220277874 rs146221748 Human genes 0.000 description 3
- 102220313606 rs1553373926 Human genes 0.000 description 3
- 102200142777 rs1554112524 Human genes 0.000 description 3
- 239000007858 starting material Substances 0.000 description 3
- 230000009466 transformation Effects 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- 239000013598 vector Substances 0.000 description 3
- IYLLULUTZPKQBW-UHFFFAOYSA-N Acrinol Chemical compound CC(O)C(O)=O.C1=C(N)C=CC2=C(N)C3=CC(OCC)=CC=C3N=C21 IYLLULUTZPKQBW-UHFFFAOYSA-N 0.000 description 2
- 108010062877 Bacteriocins Proteins 0.000 description 2
- 108010022366 Carcinoembryonic Antigen Proteins 0.000 description 2
- 102100025475 Carcinoembryonic antigen-related cell adhesion molecule 5 Human genes 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 238000001712 DNA sequencing Methods 0.000 description 2
- 102100036242 HLA class II histocompatibility antigen, DQ alpha 2 chain Human genes 0.000 description 2
- 101000930801 Homo sapiens HLA class II histocompatibility antigen, DQ alpha 2 chain Proteins 0.000 description 2
- 101001030609 Homo sapiens Mucin-like protein 3 Proteins 0.000 description 2
- 108700005091 Immunoglobulin Genes Proteins 0.000 description 2
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 description 2
- 102000017727 Immunoglobulin Variable Region Human genes 0.000 description 2
- 108010036940 Levansucrase Proteins 0.000 description 2
- PEEHTFAAVSWFBL-UHFFFAOYSA-N Maleimide Chemical compound O=C1NC(=O)C=C1 PEEHTFAAVSWFBL-UHFFFAOYSA-N 0.000 description 2
- 102000018697 Membrane Proteins Human genes 0.000 description 2
- 108010052285 Membrane Proteins Proteins 0.000 description 2
- 108010006519 Molecular Chaperones Proteins 0.000 description 2
- 102000005431 Molecular Chaperones Human genes 0.000 description 2
- 102100038572 Mucin-like protein 3 Human genes 0.000 description 2
- 101150056612 PPIA gene Proteins 0.000 description 2
- 108010090127 Periplasmic Proteins Proteins 0.000 description 2
- 108010034634 Repressor Proteins Proteins 0.000 description 2
- 102000009661 Repressor Proteins Human genes 0.000 description 2
- 108010039491 Ricin Proteins 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 108091081024 Start codon Proteins 0.000 description 2
- 239000004098 Tetracycline Substances 0.000 description 2
- 102100022153 Tumor necrosis factor receptor superfamily member 4 Human genes 0.000 description 2
- 239000011543 agarose gel Substances 0.000 description 2
- DLRVVLDZNNYCBX-CAPXFGMSSA-N allolactose Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](O)O1 DLRVVLDZNNYCBX-CAPXFGMSSA-N 0.000 description 2
- 238000005571 anion exchange chromatography Methods 0.000 description 2
- 230000003115 biocidal effect Effects 0.000 description 2
- 238000006664 bond formation reaction Methods 0.000 description 2
- 239000013592 cell lysate Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 2
- 239000013611 chromosomal DNA Substances 0.000 description 2
- 230000004186 co-expression Effects 0.000 description 2
- 239000013068 control sample Substances 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000011143 downstream manufacturing Methods 0.000 description 2
- 101150015101 dsbC gene Proteins 0.000 description 2
- 230000009977 dual effect Effects 0.000 description 2
- 229960004189 ethacridine lactate Drugs 0.000 description 2
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 2
- 229960005542 ethidium bromide Drugs 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 229910001385 heavy metal Inorganic materials 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 108010044426 integrins Proteins 0.000 description 2
- 102000006495 integrins Human genes 0.000 description 2
- 231100000518 lethal Toxicity 0.000 description 2
- 230000001665 lethal effect Effects 0.000 description 2
- 239000012160 loading buffer Substances 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 210000004962 mammalian cell Anatomy 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000036961 partial effect Effects 0.000 description 2
- 229920001281 polyalkylene Polymers 0.000 description 2
- 101150108030 ppiD gene Proteins 0.000 description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 230000006798 recombination Effects 0.000 description 2
- 238000005215 recombination Methods 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 108091008146 restriction endonucleases Proteins 0.000 description 2
- 238000012552 review Methods 0.000 description 2
- 101150025220 sacB gene Proteins 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 229960002180 tetracycline Drugs 0.000 description 2
- 229930101283 tetracycline Natural products 0.000 description 2
- 235000019364 tetracycline Nutrition 0.000 description 2
- 150000003522 tetracyclines Chemical class 0.000 description 2
- 150000003573 thiols Chemical class 0.000 description 2
- 230000001960 triggered effect Effects 0.000 description 2
- 210000005253 yeast cell Anatomy 0.000 description 2
- 102100031585 ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 Human genes 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 229920000856 Amylose Polymers 0.000 description 1
- 102100031930 Anterior gradient protein 3 Human genes 0.000 description 1
- 101100136076 Aspergillus oryzae (strain ATCC 42149 / RIB 40) pel1 gene Proteins 0.000 description 1
- 102100027705 Astrotactin-2 Human genes 0.000 description 1
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 1
- 102100022005 B-lymphocyte antigen CD20 Human genes 0.000 description 1
- 101000870242 Bacillus phage Nf Tail knob protein gp9 Proteins 0.000 description 1
- 231100000699 Bacterial toxin Toxicity 0.000 description 1
- 108010078959 C-terminal processing peptidase Proteins 0.000 description 1
- 102100024217 CAMPATH-1 antigen Human genes 0.000 description 1
- 102100031173 CCN family member 4 Human genes 0.000 description 1
- 101710137353 CCN family member 4 Proteins 0.000 description 1
- 108010046080 CD27 Ligand Proteins 0.000 description 1
- 108010029697 CD40 Ligand Proteins 0.000 description 1
- 101150013553 CD40 gene Proteins 0.000 description 1
- 102100032937 CD40 ligand Human genes 0.000 description 1
- 102100025221 CD70 antigen Human genes 0.000 description 1
- 108091011896 CSF1 Proteins 0.000 description 1
- 102100035350 CUB domain-containing protein 1 Human genes 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 108010071942 Colony-Stimulating Factors Proteins 0.000 description 1
- 241000701022 Cytomegalovirus Species 0.000 description 1
- 101710197960 D-aminoacyl-tRNA deacylase Proteins 0.000 description 1
- 102100029010 D-aminoacyl-tRNA deacylase 1 Human genes 0.000 description 1
- 101710109959 D-aminoacyl-tRNA deacylase 1 Proteins 0.000 description 1
- 108010037897 DC-specific ICAM-3 grabbing nonintegrin Proteins 0.000 description 1
- 239000003155 DNA primer Substances 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 102100038027 Diacylglycerol lipase-alpha Human genes 0.000 description 1
- 241000588700 Dickeya chrysanthemi Species 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 108010024212 E-Selectin Proteins 0.000 description 1
- 102100023471 E-selectin Human genes 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
- 238000002965 ELISA Methods 0.000 description 1
- 102100025137 Early activation antigen CD69 Human genes 0.000 description 1
- 108010055325 EphB3 Receptor Proteins 0.000 description 1
- 102100031982 Ephrin type-B receptor 3 Human genes 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 108010017080 Granulocyte Colony-Stimulating Factor Proteins 0.000 description 1
- 102000004269 Granulocyte Colony-Stimulating Factor Human genes 0.000 description 1
- 102000004457 Granulocyte-Macrophage Colony-Stimulating Factor Human genes 0.000 description 1
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 1
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 101000777636 Homo sapiens ADP-ribosyl cyclase/cyclic ADP-ribose hydrolase 1 Proteins 0.000 description 1
- 101000775037 Homo sapiens Anterior gradient protein 3 Proteins 0.000 description 1
- 101000936743 Homo sapiens Astrotactin-2 Proteins 0.000 description 1
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 1
- 101000897405 Homo sapiens B-lymphocyte antigen CD20 Proteins 0.000 description 1
- 101000980814 Homo sapiens CAMPATH-1 antigen Proteins 0.000 description 1
- 101000737742 Homo sapiens CUB domain-containing protein 1 Proteins 0.000 description 1
- 101000950954 Homo sapiens Diacylglycerol lipase-alpha Proteins 0.000 description 1
- 101000934374 Homo sapiens Early activation antigen CD69 Proteins 0.000 description 1
- 101000606465 Homo sapiens Inactive tyrosine-protein kinase 7 Proteins 0.000 description 1
- 101001046686 Homo sapiens Integrin alpha-M Proteins 0.000 description 1
- 101000935040 Homo sapiens Integrin beta-2 Proteins 0.000 description 1
- 101001057504 Homo sapiens Interferon-stimulated gene 20 kDa protein Proteins 0.000 description 1
- 101000998146 Homo sapiens Interleukin-17A Proteins 0.000 description 1
- 101000998151 Homo sapiens Interleukin-17F Proteins 0.000 description 1
- 101001055144 Homo sapiens Interleukin-2 receptor subunit alpha Proteins 0.000 description 1
- 101001054649 Homo sapiens Latent-transforming growth factor beta-binding protein 2 Proteins 0.000 description 1
- 101001054646 Homo sapiens Latent-transforming growth factor beta-binding protein 3 Proteins 0.000 description 1
- 101001039236 Homo sapiens Leucine-rich repeat and fibronectin type-III domain-containing protein 2 Proteins 0.000 description 1
- 101000878605 Homo sapiens Low affinity immunoglobulin epsilon Fc receptor Proteins 0.000 description 1
- 101000934338 Homo sapiens Myeloid cell surface antigen CD33 Proteins 0.000 description 1
- 101001017783 Homo sapiens Protein LRATD2 Proteins 0.000 description 1
- 101000962438 Homo sapiens Protein MAL2 Proteins 0.000 description 1
- 101000693049 Homo sapiens Protein S100-A14 Proteins 0.000 description 1
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 1
- 101001100101 Homo sapiens Retinoic acid-induced protein 3 Proteins 0.000 description 1
- 101000800616 Homo sapiens Teneurin-3 Proteins 0.000 description 1
- 101000831862 Homo sapiens Transmembrane protein 45B Proteins 0.000 description 1
- 101000679851 Homo sapiens Tumor necrosis factor receptor superfamily member 4 Proteins 0.000 description 1
- 101000851370 Homo sapiens Tumor necrosis factor receptor superfamily member 9 Proteins 0.000 description 1
- 101000899435 Homo sapiens Uncharacterized protein C1orf159 Proteins 0.000 description 1
- 229920000869 Homopolysaccharide Polymers 0.000 description 1
- 102000018071 Immunoglobulin Fc Fragments Human genes 0.000 description 1
- 108010091135 Immunoglobulin Fc Fragments Proteins 0.000 description 1
- 102100039813 Inactive tyrosine-protein kinase 7 Human genes 0.000 description 1
- 102100022338 Integrin alpha-M Human genes 0.000 description 1
- 108010008212 Integrin alpha4beta1 Proteins 0.000 description 1
- 102100025390 Integrin beta-2 Human genes 0.000 description 1
- 102000006992 Interferon-alpha Human genes 0.000 description 1
- 108010047761 Interferon-alpha Proteins 0.000 description 1
- 108090000467 Interferon-beta Proteins 0.000 description 1
- 102000003996 Interferon-beta Human genes 0.000 description 1
- 108010074328 Interferon-gamma Proteins 0.000 description 1
- 102000008070 Interferon-gamma Human genes 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 102000013462 Interleukin-12 Human genes 0.000 description 1
- 108010065805 Interleukin-12 Proteins 0.000 description 1
- 102000003816 Interleukin-13 Human genes 0.000 description 1
- 108090000176 Interleukin-13 Proteins 0.000 description 1
- 102000013691 Interleukin-17 Human genes 0.000 description 1
- 108050003558 Interleukin-17 Proteins 0.000 description 1
- 102100033461 Interleukin-17A Human genes 0.000 description 1
- 102100033454 Interleukin-17F Human genes 0.000 description 1
- 102000000588 Interleukin-2 Human genes 0.000 description 1
- 108010002350 Interleukin-2 Proteins 0.000 description 1
- 102100026878 Interleukin-2 receptor subunit alpha Human genes 0.000 description 1
- 102000000646 Interleukin-3 Human genes 0.000 description 1
- 108010002386 Interleukin-3 Proteins 0.000 description 1
- 102000004388 Interleukin-4 Human genes 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 102100039897 Interleukin-5 Human genes 0.000 description 1
- 108010002616 Interleukin-5 Proteins 0.000 description 1
- 102000004889 Interleukin-6 Human genes 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 102000010782 Interleukin-7 Receptors Human genes 0.000 description 1
- 108010038498 Interleukin-7 Receptors Proteins 0.000 description 1
- 108090001007 Interleukin-8 Proteins 0.000 description 1
- 102000004890 Interleukin-8 Human genes 0.000 description 1
- 102000015696 Interleukins Human genes 0.000 description 1
- 108010063738 Interleukins Proteins 0.000 description 1
- 108010092694 L-Selectin 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
- 102100033467 L-selectin Human genes 0.000 description 1
- 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 1
- 235000019687 Lamb Nutrition 0.000 description 1
- 102100027017 Latent-transforming growth factor beta-binding protein 2 Human genes 0.000 description 1
- 102100040698 Leucine-rich repeat and fibronectin type-III domain-containing protein 2 Human genes 0.000 description 1
- 102100038007 Low affinity immunoglobulin epsilon Fc receptor Human genes 0.000 description 1
- 102000004083 Lymphotoxin-alpha Human genes 0.000 description 1
- 108090000542 Lymphotoxin-alpha Proteins 0.000 description 1
- 108010058398 Macrophage Colony-Stimulating Factor Receptor Proteins 0.000 description 1
- 102100028123 Macrophage colony-stimulating factor 1 Human genes 0.000 description 1
- 102100028198 Macrophage colony-stimulating factor 1 receptor Human genes 0.000 description 1
- 102000047724 Member 2 Solute Carrier Family 12 Human genes 0.000 description 1
- 102100037653 Metalloreductase STEAP3 Human genes 0.000 description 1
- 101710147238 Metalloreductase STEAP3 Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 229930192392 Mitomycin Natural products 0.000 description 1
- MSFSPUZXLOGKHJ-UHFFFAOYSA-N Muraminsaeure Natural products OC(=O)C(C)OC1C(N)C(O)OC(CO)C1O MSFSPUZXLOGKHJ-UHFFFAOYSA-N 0.000 description 1
- 102100025243 Myeloid cell surface antigen CD33 Human genes 0.000 description 1
- NWIBSHFKIJFRCO-WUDYKRTCSA-N Mytomycin Chemical compound C1N2C(C(C(C)=C(N)C3=O)=O)=C3[C@@H](COC(N)=O)[C@@]2(OC)[C@@H]2[C@H]1N2 NWIBSHFKIJFRCO-WUDYKRTCSA-N 0.000 description 1
- 101710095135 NAD(P)H dehydrogenase [quinone] 1 Proteins 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108700006385 OmpF Proteins 0.000 description 1
- 108010035766 P-Selectin Proteins 0.000 description 1
- 102100023472 P-selectin Human genes 0.000 description 1
- 238000009004 PCR Kit Methods 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 108010013639 Peptidoglycan Proteins 0.000 description 1
- 108090000316 Pitrilysin Proteins 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 241000276498 Pollachius virens Species 0.000 description 1
- 108010076039 Polyproteins Proteins 0.000 description 1
- 241001415846 Procellariidae Species 0.000 description 1
- 101710089372 Programmed cell death protein 1 Proteins 0.000 description 1
- 102100033355 Protein LRATD2 Human genes 0.000 description 1
- 102100039191 Protein MAL2 Human genes 0.000 description 1
- 102100026298 Protein S100-A14 Human genes 0.000 description 1
- 101100084022 Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1) lapA gene Proteins 0.000 description 1
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 1
- 241000725643 Respiratory syncytial virus Species 0.000 description 1
- 102100038453 Retinoic acid-induced protein 3 Human genes 0.000 description 1
- 102000018673 SEC Translocation Channels Human genes 0.000 description 1
- 108010091732 SEC Translocation Channels Proteins 0.000 description 1
- 108091006620 SLC12A2 Proteins 0.000 description 1
- 101100420762 Schizophyllum commune SC6 gene Proteins 0.000 description 1
- 108091003202 SecA Proteins Proteins 0.000 description 1
- 241000270295 Serpentes Species 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 102100033191 Teneurin-3 Human genes 0.000 description 1
- 102100024181 Transmembrane protein 45B Human genes 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 102100024587 Tumor necrosis factor ligand superfamily member 15 Human genes 0.000 description 1
- 108090000138 Tumor necrosis factor ligand superfamily member 15 Proteins 0.000 description 1
- 101710165473 Tumor necrosis factor receptor superfamily member 4 Proteins 0.000 description 1
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 description 1
- 102100036856 Tumor necrosis factor receptor superfamily member 9 Human genes 0.000 description 1
- 102100022520 Uncharacterized protein C1orf159 Human genes 0.000 description 1
- 108010053099 Vascular Endothelial Growth Factor Receptor-2 Proteins 0.000 description 1
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 1
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 1
- 241000700605 Viruses Species 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
- 238000009825 accumulation Methods 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 150000003978 alpha-halocarboxylic acids Chemical class 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 238000011091 antibody purification Methods 0.000 description 1
- 210000000628 antibody-producing cell Anatomy 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 229940034982 antineoplastic agent Drugs 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 125000004429 atom Chemical group 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 239000000688 bacterial toxin Substances 0.000 description 1
- 230000009286 beneficial effect Effects 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
- 230000000903 blocking effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 102220361446 c.398A>C Human genes 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 239000012560 cell impurity Substances 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 238000012993 chemical processing Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 230000021615 conjugation Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000007822 coupling agent Substances 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000011026 diafiltration Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- AFOSIXZFDONLBT-UHFFFAOYSA-N divinyl sulfone Chemical compound C=CS(=O)(=O)C=C AFOSIXZFDONLBT-UHFFFAOYSA-N 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 238000000804 electron spin resonance spectroscopy Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000012239 gene modification Methods 0.000 description 1
- 230000005017 genetic modification Effects 0.000 description 1
- 235000013617 genetically modified food Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 229920006158 high molecular weight polymer Polymers 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 235000020256 human milk Nutrition 0.000 description 1
- 210000004251 human milk Anatomy 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 150000003949 imides Chemical class 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000016784 immunoglobulin production Effects 0.000 description 1
- 230000003116 impacting effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 206010022000 influenza Diseases 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 229960003130 interferon gamma Drugs 0.000 description 1
- 229960001388 interferon-beta Drugs 0.000 description 1
- 229940047122 interleukins Drugs 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- XMBWDFGMSWQBCA-RNFDNDRNSA-M iodine-131(1-) Chemical compound [131I-] XMBWDFGMSWQBCA-RNFDNDRNSA-M 0.000 description 1
- PGLTVOMIXTUURA-UHFFFAOYSA-N iodoacetamide Chemical compound NC(=O)CI PGLTVOMIXTUURA-UHFFFAOYSA-N 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 150000002597 lactoses Chemical class 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 229920006008 lipopolysaccharide Polymers 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 150000002678 macrocyclic compounds Chemical class 0.000 description 1
- 125000005439 maleimidyl group Chemical group C1(C=CC(N1*)=O)=O 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
- 238000001471 micro-filtration Methods 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 229960004857 mitomycin Drugs 0.000 description 1
- 238000001823 molecular biology technique Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 101150040383 pel2 gene Proteins 0.000 description 1
- 101150050446 pelB gene Proteins 0.000 description 1
- 230000008823 permeabilization Effects 0.000 description 1
- 101150009573 phoA gene Proteins 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 229920002859 polyalkenylene Polymers 0.000 description 1
- 229920001451 polypropylene glycol Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 229920002451 polyvinyl alcohol Polymers 0.000 description 1
- 230000004481 post-translational protein modification Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000013615 primer Substances 0.000 description 1
- 238000011027 product recovery Methods 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 238000000159 protein binding assay Methods 0.000 description 1
- 230000006920 protein precipitation Effects 0.000 description 1
- 238000002708 random mutagenesis Methods 0.000 description 1
- 238000010223 real-time analysis Methods 0.000 description 1
- 125000006853 reporter group Chemical group 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 230000009962 secretion pathway Effects 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 230000037432 silent mutation Effects 0.000 description 1
- 239000003998 snake venom Substances 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 239000002708 spider venom Substances 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 125000004434 sulfur atom Chemical group 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/18—Ion-exchange chromatography
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/16—Extraction; Separation; Purification by chromatography
- C07K1/22—Affinity chromatography or related techniques based upon selective absorption processes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/30—Extraction; Separation; Purification by precipitation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/475—Growth factors; Growth regulators
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
- C07K16/241—Tumor Necrosis Factors
-
- 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
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/10—Immunoglobulins specific features characterized by their source of isolation or production
- C07K2317/14—Specific host cells or culture conditions, e.g. components, pH or temperature
-
- 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'
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biomedical Technology (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
- Peptides Or Proteins (AREA)
Description
a.宿主細胞試料又はその抽出物のpHをpH5以下に調整して、組換えジスルフィドイソメラーゼを沈殿させるステップ、及び
b.沈殿した組換えジスルフィドイソメラーゼを組換えタンパク質から分離して、組換えタンパク質試料を提供するステップ
を含む方法が提供される。
a.宿主細胞試料又はその抽出物のpHをpH範囲3.5から5に調整して、組換えDsbCを沈殿させるステップ、及び
b.沈殿した組換えDsbCを分離して、精製された組換え抗体又はその断片を含有する試料を提供するステップ
を含む方法が提供される。
配列番号1は、CDP870のCDRH1のアミノ酸配列を示す。
・ 細胞から一部若しくは全ての化学的物質を抽出するための部分的又は全細胞溶解から得られる材料、又は
・ 細胞から発現されるタンパク質の1つ又は複数が液相内に放出されるように細胞をプロセシングすることから得られる材料
を言うために用いられる。
・ FkpA、Skp、SurA、PPiA、及びPPiDなどの、タンパク質のフォールディングを促進し得る1つ又は複数のタンパク質、並びに/又は
・ SecY、SecE、SecG、SecYEG、SecA、SecB、FtsY、及びLepなどの、タンパク質の分泌若しくは転位を促進し得る1つ又は複数のタンパク質、並びに/又は
・ DsbA、DsbB、DsbD、DsbGなどの、ジスルフィド結合の形成を促進し得る1つ又は複数のタンパク質。
・ FkpA、Skp、SurA、PPiA、及びPPiDなどの、タンパク質のフォールディングを促進し得る1つ又は複数のタンパク質、
・ SecY、SecE、SecG、SecYEG、SecA、SecB、FtsY、及びLepなどの、タンパク質の分泌若しくは転位を促進し得る1つ又は複数のタンパク質、並びに
・ DsbA、DsbB、DsbD、DsbGなどの、ジスルフィド結合の形成を促進し得る1つ又は複数のタンパク質。
・ 組換えタンパク質及び組換えジスルフィドイソメラーゼを発現する宿主細胞の集団を含む溶液、
・ 宿主細胞を除去するための遠心分離及び/又は濾過の後の、組換えタンパク質及び組換えジスルフィドイソメラーゼを発現する宿主細胞の集団の上清、
・ 熱処理などの抽出ステップの後の、組換えタンパク質及び組換えジスルフィドイソメラーゼを発現する宿主細胞の集団の抽出物、或いは
・ 熱処理などの抽出ステップ並びに1つ又は複数のその後の遠心分離及び/又は濾過のステップの後の、組換えタンパク質及び組換えジスルフィドイソメラーゼを発現する宿主細胞の集団の抽出物
に対して行うことができる。
組換えDsbC及び組換え抗体を発現する宿主細胞系の生成
全ての実験で、大腸菌細胞系W3110を親野生型細胞系として用いた。
a.突然変異したTsp遺伝子、
b.突然変異したTsp遺伝子及び運搬組換えDsbC、
c.突然変異したTsp遺伝子及び突然変異したspr遺伝子、
d.突然変異したTsp遺伝子及び突然変異したspr遺伝子及び運搬組換えDsbC。
MXE001株を以下のように生成した:
Tspカセットを、SalI制限断片、NotI制限断片として、同様に制限されたpKO3プラスミド内に移動させた。pKO3プラスミドは、染色体組み込み事象を起こして選択するためのクロラムフェニコールマーカーと共に、pSC101複製起点(RepA)の温度感受性の突然変異体を用いる。レバンスクラーゼをコードするsacB遺伝子は、ショ糖上で成長した大腸菌に対して致死的であり、したがって、(クロラムフェニコールマーカー及びpSC101起点と共に)脱組み込み事象及びプラスミドキュアリング事象を起こして選択するために用いられる。この方法論は、以前に記載されている(Hamilton et al., 1989, Journal of Bacteriology, 171, 4617-4622、及びBlomfield et al., 1991, Molecular Microbiology, 5, 1447-1457)。pKO3系は、挿入された遺伝子を除いて全ての選択マーカーを宿主ゲノムから除去する。
EcoRI制限マーカー及びAseI制限マーカーを含む、配列番号15で示されるノックアウト突然変異したTsp遺伝子を含むpMXE191。
5ul 緩衝液×10(Roche)
1ul dNTPミックス(Roche、10mMミックス)
1.5ul 5’オリゴ(5pmol)
1.5ul 3’オリゴ(5pmol)
2ul 細胞溶解物
0.5ul TaqDNAポリメラーゼ(Roche 5U/ul)
38.5ul H2O
PCRサイクル
94℃ 1分
94℃ 1分)
55℃ 1分)30サイクル反復
72℃ 1分)
72℃ 10分
spr突然変異を生じさせ、相補性アッセイの使用に選択した。
1.V98E
2.D133A
3.V135D
4.V135G
5.G147C
6.S95F及びY115F
pMXE336、pK03 spr S95F(−SalI)
pMXE337、pK03 spr Y115F(−SalI)
pMXE338、pK03 spr G147C(−SalI)
pMXE339、pK03 spr D133A(−SalI)
pMXE340、pK03 spr V135D (−SalI)
pMXE341、pK03 spr V135G(−SalI)
pMXE342、pK03 spr V98E(−SalI)
pMXE343、pK03 spr C94A(−SalI)
pMXE344、pK03 spr H145A(−SalI)
pMXE345、pK03 spr H157A(−SalI)
pMXE346、pK03 spr W174R(−SalI)
抗TNF Fab’の重鎖配列及び軽鎖配列とDsbCをコードする配列との両方を含有するプラスミドを構築した。
MXE008株を、Fab’軽鎖、Fab’重鎖、及びDsbCをコードするプラスミドで形質転換した。
株MXE008を、Sambrook et al 1989, Molecular cloning: a laboratory manual. CSHL press, N.Y.において見ることができる従来の制限クローニング方法を用いて構築された、CDP870のFab’(配列番号9で示される軽鎖配列及び配列番号10で示される重鎖配列を有する抗TNF Fab’)の発現ベクターである、プラスミドpMXE117(pTTO CDP870又は40.4 IGS2)で形質転換した。プラスミドpMXE117(pTTO CDP870又は40.4 IGS2)は、強力なtacプロモーター配列及びlacオペレーター配列という特徴を含んでいた。Fabの軽鎖遺伝子及び重鎖遺伝子を、単一のジシストロン性メッセージとして転写した。大腸菌OmpAタンパク質に由来するシグナルペプチドをコードするDNAを、軽鎖遺伝子配列及び重鎖遺伝子配列の両方の5’末端に融合し、これは、大腸菌ペリプラズムへのポリペプチドの転位を指示した。転写を、二重転写ターミネーターrrnB t1t2を用いて終結させた。lacIq遺伝子は、構成的に発現したLacIリプレッサータンパク質をコードした。これは、脱抑制がアロラクトース又はIPTGの存在によって誘発されるまで、tacプロモーターからの転写を抑制した。用いられた複製起点はp15Aであり、これは、少ないコピー数を維持した。プラスミドは、抗生物質の選択のためのテトラサイクリン耐性遺伝子を含有していた。
組換え抗体抗TNF Fab’を発現する宿主細胞の成長
例1において生産されたMXE008株を、抗TNFαFab’を発現させるための発酵実験において用いた。
大腸菌株からの抗TNFαFab’の抽出及びpH調整
例2からの凍結ペレット細胞ペレットを次いで解凍し、抽出緩衝液(0.1MのTris/10mMのEDTA、pH7.4)内に再懸濁させた。抽出は、60℃で一晩行った。
逆相HPLCによるpH調整の後の宿主細胞試料抽出物の分析
pH5.0、4.5、若しくは3.0へのpH調整の後の各宿主細胞試料抽出物試料、又は対照試料(pH調整なし、pH7)を、時点(T)=0、T=1時間、T=2時間、T=4.5時間、T=8時間、及びT=24時間の時間経過にわたって採取し、リアルタイムの分析を、逆相HPLCを介して行った。逆相HPLCは、疎水性に基づくタンパク質の分離を可能にし、適切なHPLCカラム(Agilent Technologiesの、Poroshell(商標)300SB−C8、5ミクロン、2.1×75mm)を用いて行われた。試料を分析の前に濾過した(0.22um)。
試料のSDS−PAGE分析
非還元SDS−PAGE分析を、pH7、pH5.0、pH4.5、及びpH3.0での長期保存の後に行った。図7は、SDS−PAGE分析ゲルを示す。図7から、DBPの量が、pH3へのpH調整の後に低減していることが分かる。DsbC及びFab’の重鎖(HC)は、ゲル上で同一の分子量を示すが、DsbC及びHCのバンドが、DsbCの低減に起因して、pH5、pH4.5、及びpH3へのpH調整の後に低減していることが分かり、このことにより、DsbC−ヒス及びDBPの量がpH調整後に低減していることが確認される。
Claims (21)
- pI6以上を有する組換えタンパク質をグラム陰性細菌宿主細胞のバッチ又はその抽出物から精製するための方法であって、前記宿主細胞が、組換えタンパク質及び組換えジスルフィドイソメラーゼDsbCを発現し、
a.宿主細胞のバッチ又はその抽出物のpHをpH5〜3に調整して、組換えジスルフィドイソメラーゼを沈殿させるステップ、及び
b.沈殿した組換えジスルフィドイソメラーゼを組換えタンパク質から分離して、組換えタンパク質試料を生産するステップ
を含む上記方法。 - 宿主細胞のバッチ又はその抽出物のpHがpH4.5から3.0に調整される、請求項1に記載の方法。
- ジスルフィドイソメラーゼがヒスチジンタグを含む、請求項1又は2に記載の方法。
- グラム陰性細菌宿主細胞が大腸菌(E.coli)である、請求項1から3までのいずれか一項に記載の方法。
- 宿主細胞がFkpa及び/又はSkpを含む、請求項1から4までのいずれか一項に記載の方法。
- ステップa)において、宿主細胞試料のバッチ又はその抽出物のpHが、氷酢酸を用いてpH5〜3に調整される、請求項1から5までのいずれか一項に記載の方法。
- 宿主細胞のバッチ又はその抽出物がpH5〜3で1時間以下にわたり保持される、請求項1から6までのいずれか一項に記載の方法。
- ステップb)における分離が遠心分離及び/又は濾過を含む、請求項1から7までのいずれか一項に記載の方法。
- ステップb)の後に、組換えタンパク質をクロマトグラフィーに供するさらなる精製ステップを含む、請求項1から8までのいずれか一項に記載の方法。
- クロマトグラフィーがイオン交換クロマトグラフィーである、請求項9に記載の方法。
- ステップb)の後に、組換えタンパク質のpHがpH5から7に調整される、請求項1から10までのいずれか一項に記載の方法。
- ステップa)の前に、抽出緩衝液が宿主細胞のバッチ又はその抽出物に添加され、宿主細胞のバッチ又はその抽出物が熱処理ステップに供される、請求項1から11までのいずれか一項に記載の方法。
- 組換えタンパク質がpI6から9を有する、請求項1から12までのいずれか一項に記載の方法。
- 組換えタンパク質がpI8から9を有する、請求項13に記載の方法。
- 組換えタンパク質が組換え抗体である、請求項1から14までのいずれか一項に記載の方法。
- 組換え抗体が、モノクローナル抗体、ヒト化抗体、若しくはキメラ抗体、又はその断片、例えばその結合断片である、請求項15に記載の方法。
- 組換え抗体がFab断片又はFab’断片である、請求項15又は請求項16に記載の方法。
- 組換え抗体がヒトTNFαに特異的である、請求項15から17までのいずれか一項に記載の方法。
- 抗体が、可変ドメインがCDRH1について配列番号1、CDRH2について配列番号2、及びCDRH3について配列番号3で示される配列を有するCDRを含む重鎖、並びに可変ドメインがCDRL1について配列番号4、CDRL2について配列番号5、及びCDRL3について配列番号6で示される配列を有するCDRを含む軽鎖を含む、請求項18に記載の方法。
- 抗体が、配列番号7で示される軽鎖可変領域配列及び配列番号8で示される重鎖可変領域を含む、請求項19に記載の方法。
- 抗体がFab断片であり、配列番号10で示される配列を有する重鎖及び配列番号9で示される配列を有する軽鎖を含む、請求項20に記載の方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
GBGB1012599.5A GB201012599D0 (en) | 2010-07-27 | 2010-07-27 | Process for purifying proteins |
GB1012599.5 | 2010-07-27 | ||
PCT/GB2011/001129 WO2012013930A2 (en) | 2010-07-27 | 2011-07-27 | Process for purifying proteins |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2013535463A JP2013535463A (ja) | 2013-09-12 |
JP5935222B2 true JP5935222B2 (ja) | 2016-06-15 |
Family
ID=42752868
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2013521208A Active JP5935222B2 (ja) | 2010-07-27 | 2011-07-27 | タンパク質を精製するプロセス |
Country Status (24)
Country | Link |
---|---|
US (1) | US9751930B2 (ja) |
EP (1) | EP2598516B8 (ja) |
JP (1) | JP5935222B2 (ja) |
KR (1) | KR101944121B1 (ja) |
CN (1) | CN103189385B (ja) |
AU (1) | AU2011284548B2 (ja) |
BR (1) | BR112013000177B1 (ja) |
CA (1) | CA2804099C (ja) |
CY (1) | CY1116526T1 (ja) |
DK (1) | DK2598516T3 (ja) |
ES (1) | ES2537877T3 (ja) |
GB (1) | GB201012599D0 (ja) |
HR (1) | HRP20150639T1 (ja) |
HU (1) | HUE026049T2 (ja) |
IL (1) | IL224044A (ja) |
ME (1) | ME02162B (ja) |
MX (1) | MX337184B (ja) |
PL (1) | PL2598516T3 (ja) |
PT (1) | PT2598516E (ja) |
RS (1) | RS54085B1 (ja) |
SG (2) | SG186974A1 (ja) |
SI (1) | SI2598516T1 (ja) |
SM (1) | SMT201500142B (ja) |
WO (1) | WO2012013930A2 (ja) |
Families Citing this family (14)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU2010299640B2 (en) | 2009-09-24 | 2016-02-25 | Ucb Pharma S.A. | Bacterial host strain |
GB201000591D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist strain |
GB201000587D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist strain |
GB201000590D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial host strain |
GB201001791D0 (en) | 2010-02-03 | 2010-03-24 | Ucb Pharma Sa | Process for obtaining antibodies |
EP3339325A1 (en) | 2011-07-13 | 2018-06-27 | UCB Biopharma SPRL | Bacterial host strain expressing recombinant dsbc |
GB201208367D0 (en) * | 2012-05-14 | 2012-06-27 | Ucb Pharma Sa | Biological product |
CN104507955A (zh) * | 2012-05-31 | 2015-04-08 | 新加坡科技研究局 | 采用具有多形式官能性的颗粒对免疫球蛋白g制备物的色谱纯化 |
WO2013180648A1 (en) * | 2012-05-31 | 2013-12-05 | Agency For Science, Technology And Research | Methods for use of mixed multifunctional surfaces for reducing aggregate content in protein preparations |
EA039002B1 (ru) | 2014-12-22 | 2021-11-19 | Юсб Биофарма Спрл | Способ получения белка |
HRP20231408T1 (hr) | 2014-12-22 | 2024-02-16 | UCB Biopharma SRL | Proizvodnja proteina |
SI3265557T1 (sl) * | 2015-03-06 | 2020-02-28 | F. Hoffmann-La Roche Ag | Ultraprečiščena DsbA in DsbC ter postopki njune izdelave in uporabe |
CN110272491B (zh) * | 2018-03-13 | 2023-01-24 | 江苏恒瑞医药股份有限公司 | 一种抗pd-1抗体的纯化工艺 |
EP4010500A1 (de) * | 2019-08-05 | 2022-06-15 | Wacker Chemie AG | Bakterienstamm zur freisetzung eines rekombinanten proteins in einem fermentationsverfahren |
Family Cites Families (57)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
GB8422238D0 (en) | 1984-09-03 | 1984-10-10 | Neuberger M S | Chimeric proteins |
GB8719042D0 (en) | 1987-08-12 | 1987-09-16 | Parker D | Conjugate compounds |
US5030570A (en) | 1988-06-30 | 1991-07-09 | The Eunice Kennedy Shriver Center For Mental Retardation | DNA encoding and method of expressing human monoamine oxidase type A |
US5530101A (en) | 1988-12-28 | 1996-06-25 | Protein Design Labs, Inc. | Humanized immunoglobulins |
GB8907617D0 (en) | 1989-04-05 | 1989-05-17 | Celltech Ltd | Drug delivery system |
GB8928874D0 (en) | 1989-12-21 | 1990-02-28 | Celltech Ltd | Humanised antibodies |
CA2090126C (en) | 1990-08-02 | 2002-10-22 | John W. Schrader | Methods for the production of proteins with a desired function |
US5264365A (en) | 1990-11-09 | 1993-11-23 | Board Of Regents, The University Of Texas System | Protease-deficient bacterial strains for production of proteolytically sensitive polypeptides |
US5508192A (en) | 1990-11-09 | 1996-04-16 | Board Of Regents, The University Of Texas System | Bacterial host strains for producing proteolytically sensitive polypeptides |
GB9112536D0 (en) | 1991-06-11 | 1991-07-31 | Celltech Ltd | Chemical compounds |
GB9113120D0 (en) | 1991-06-18 | 1991-08-07 | Kodak Ltd | Photographic processing apparatus |
GB9120467D0 (en) | 1991-09-26 | 1991-11-06 | Celltech Ltd | Anti-hmfg antibodies and process for their production |
WO1993008829A1 (en) | 1991-11-04 | 1993-05-13 | The Regents Of The University Of California | Compositions that mediate killing of hiv-infected cells |
GB9215540D0 (en) | 1992-07-22 | 1992-09-02 | Celltech Ltd | Protein expression system |
AU2660397A (en) * | 1996-04-05 | 1997-10-29 | Board Of Regents, The University Of Texas System | Methods for producing soluble, biologically-active disulfide bond-containing eukaryotic proteins in bacterial cells |
GB9625640D0 (en) | 1996-12-10 | 1997-01-29 | Celltech Therapeutics Ltd | Biological products |
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 |
US6306619B1 (en) | 1999-06-29 | 2001-10-23 | Washington University | DegP periplasmic protease a new anti-infective target and an in vitro assay for DegP protease function |
GB0006398D0 (en) | 2000-03-16 | 2000-05-03 | Novartis Ag | Organic compounds |
GB0013810D0 (en) | 2000-06-06 | 2000-07-26 | Celltech Chiroscience Ltd | Biological products |
AU8867501A (en) | 2000-09-01 | 2002-03-13 | Biogen Inc | Co-crystal structure of monoclonal antibody 5c8 and cd154, and use thereof in drug design |
EP1314037A2 (en) | 2000-09-01 | 2003-05-28 | Biogen, Inc. | Methods of designing and producing compounds having improved binding affinity for cd154 or other trimeric proteins |
US7041479B2 (en) * | 2000-09-06 | 2006-05-09 | The Board Of Trustess Of The Leland Stanford Junior University | Enhanced in vitro synthesis of active proteins containing disulfide bonds |
US6828121B2 (en) | 2000-12-14 | 2004-12-07 | Genentech, Inc. | Bacterial host strains |
ATE405650T1 (de) | 2000-12-14 | 2008-09-15 | Genentech Inc | Produktion von ganzen antikörpern in prokaryontischen zellen |
WO2004081026A2 (en) | 2003-06-30 | 2004-09-23 | Domantis Limited | Polypeptides |
TWI334439B (en) | 2001-08-01 | 2010-12-11 | Centocor Inc | Anti-tnf antibodies, compositions, methods and uses |
WO2003018771A2 (en) | 2001-08-27 | 2003-03-06 | Genentech, Inc. | A system for antibody expression and assembly |
GB0124317D0 (en) | 2001-10-10 | 2001-11-28 | Celltech R&D Ltd | Biological products |
EA007905B1 (ru) | 2001-11-16 | 2007-02-27 | Байоджен Айдек Инк. | Полицистронная экспрессия антител |
GB0129105D0 (en) | 2001-12-05 | 2002-01-23 | Celltech R&D Ltd | Expression control using variable intergenic sequences |
EP2366718A3 (en) | 2002-06-28 | 2012-05-02 | Domantis Limited | Ligand |
EP1578447A4 (en) * | 2002-10-31 | 2009-06-03 | Genentech Inc | METHODS AND COMPOSITIONS THAT CAN INCREASE ANTIBODY PRODUCTION |
AU2003285578B2 (en) | 2002-12-03 | 2010-07-15 | Ucb Pharma S.A. | Assay for identifying antibody producing cells |
ES2287687T3 (es) | 2003-01-09 | 2007-12-16 | Genentech, Inc. | Purificacion de polipeptidos. |
GB0303337D0 (en) | 2003-02-13 | 2003-03-19 | Celltech R&D Ltd | Biological products |
JP4667383B2 (ja) | 2003-06-13 | 2011-04-13 | バイオジェン・アイデック・エムエイ・インコーポレイテッド | アグリコシル抗cd154(cd40リガンド)抗体およびその使用 |
GB0315457D0 (en) | 2003-07-01 | 2003-08-06 | Celltech R&D Ltd | Biological products |
GB0315450D0 (en) | 2003-07-01 | 2003-08-06 | Celltech R&D Ltd | Biological products |
JP4944608B2 (ja) | 2003-07-26 | 2012-06-06 | バイオジェン・アイデック・エムエイ・インコーポレイテッド | 改良された抗原結合親和性を有する、変更された抗体 |
RS20070027A (en) | 2004-07-26 | 2008-11-28 | Biogen Idec Ma Inc., | Anti-cd154 antibodies |
JP2008511337A (ja) | 2004-09-02 | 2008-04-17 | ジェネンテック・インコーポレーテッド | ヘテロ多量体分子 |
US7563443B2 (en) | 2004-09-17 | 2009-07-21 | Domantis Limited | Monovalent anti-CD40L antibody polypeptides and compositions thereof |
GB0425534D0 (en) | 2004-11-19 | 2004-12-22 | Celltech R&D Ltd | Process for obtaining antibodies |
JP5721951B2 (ja) | 2007-03-22 | 2015-05-20 | バイオジェン アイデック マサチューセッツ インコーポレイテッド | 抗体、抗体誘導体、および抗体断片を含む、cd154に特異的に結合する結合タンパク質ならびにその使用 |
US7662587B1 (en) | 2009-03-05 | 2010-02-16 | E. I. Du Pont De Nemours And Company | Gene knockout mutations that increase peptide production |
AU2010299640B2 (en) | 2009-09-24 | 2016-02-25 | Ucb Pharma S.A. | Bacterial host strain |
US8470552B2 (en) | 2009-10-12 | 2013-06-25 | Keck Graduate Institute | Strategy to reduce lactic acid production and control PH in animal cell culture |
SI2496601T1 (sl) | 2009-11-05 | 2017-09-29 | F. Hoffmann-La Roche Ag | Tehnike in sestavek za sekrecijo heterolognih polipeptidov |
GB201000587D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist strain |
GB201000588D0 (en) * | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial host strain |
GB201000591D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial hoist strain |
GB201000590D0 (en) | 2010-01-14 | 2010-03-03 | Ucb Pharma Sa | Bacterial host strain |
GB201001791D0 (en) | 2010-02-03 | 2010-03-24 | Ucb Pharma Sa | Process for obtaining antibodies |
EP3339325A1 (en) | 2011-07-13 | 2018-06-27 | UCB Biopharma SPRL | Bacterial host strain expressing recombinant dsbc |
EP2546267A1 (en) | 2011-07-13 | 2013-01-16 | UCB Pharma S.A. | Bacterial host strain expressing recombinant DsbC |
GB201208367D0 (en) | 2012-05-14 | 2012-06-27 | Ucb Pharma Sa | Biological product |
-
2010
- 2010-07-27 GB GBGB1012599.5A patent/GB201012599D0/en not_active Ceased
-
2011
- 2011-07-27 SI SI201130516T patent/SI2598516T1/sl unknown
- 2011-07-27 ME MEP-2015-94A patent/ME02162B/me unknown
- 2011-07-27 AU AU2011284548A patent/AU2011284548B2/en active Active
- 2011-07-27 BR BR112013000177-1A patent/BR112013000177B1/pt not_active IP Right Cessation
- 2011-07-27 WO PCT/GB2011/001129 patent/WO2012013930A2/en active Application Filing
- 2011-07-27 MX MX2013000899A patent/MX337184B/es active IP Right Grant
- 2011-07-27 DK DK11745810.9T patent/DK2598516T3/da active
- 2011-07-27 SG SG2013001318A patent/SG186974A1/en unknown
- 2011-07-27 ES ES11745810.9T patent/ES2537877T3/es active Active
- 2011-07-27 CN CN201180036417.8A patent/CN103189385B/zh active Active
- 2011-07-27 RS RS20150429A patent/RS54085B1/en unknown
- 2011-07-27 EP EP11745810.9A patent/EP2598516B8/en active Active
- 2011-07-27 SG SG2014005441A patent/SG196862A1/en unknown
- 2011-07-27 PL PL11745810T patent/PL2598516T3/pl unknown
- 2011-07-27 US US13/812,350 patent/US9751930B2/en active Active
- 2011-07-27 CA CA2804099A patent/CA2804099C/en active Active
- 2011-07-27 KR KR1020137004911A patent/KR101944121B1/ko active IP Right Grant
- 2011-07-27 JP JP2013521208A patent/JP5935222B2/ja active Active
- 2011-07-27 PT PT117458109T patent/PT2598516E/pt unknown
- 2011-07-27 HU HUE11745810A patent/HUE026049T2/en unknown
-
2012
- 2012-12-31 IL IL224044A patent/IL224044A/en active IP Right Grant
-
2015
- 2015-06-15 HR HRP20150639TT patent/HRP20150639T1/hr unknown
- 2015-06-18 SM SM201500142T patent/SMT201500142B/xx unknown
- 2015-06-23 CY CY20151100535T patent/CY1116526T1/el unknown
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP5935222B2 (ja) | タンパク質を精製するプロセス | |
US11384136B2 (en) | Process for obtaining antibodies | |
KR102036399B1 (ko) | 단백질 발현용 재조합 박테리아 숙주 세포 | |
CA2584226C (en) | Process for obtaining antibodies | |
JP5960607B2 (ja) | 変異体spr遺伝子及び野生型Tsp遺伝子を含む細菌宿主系統 | |
JP6132551B2 (ja) | シャペロン活性を保持するプロテアーゼ欠損DegP並びにノックアウトTsp及びptr遺伝子を持つ、組換えタンパク質発現のための細菌株 | |
CA2582726A1 (en) | Process for obtaining antibodies |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20140725 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20150908 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20151126 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20160308 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20160329 |
|
A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A711 Effective date: 20160414 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20160422 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20160414 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 5935222 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |