JP5480558B2 - 改善された特異性を有する新規免疫グロブリン結合タンパク質 - Google Patents
改善された特異性を有する新規免疫グロブリン結合タンパク質 Download PDFInfo
- Publication number
- JP5480558B2 JP5480558B2 JP2009185389A JP2009185389A JP5480558B2 JP 5480558 B2 JP5480558 B2 JP 5480558B2 JP 2009185389 A JP2009185389 A JP 2009185389A JP 2009185389 A JP2009185389 A JP 2009185389A JP 5480558 B2 JP5480558 B2 JP 5480558B2
- Authority
- JP
- Japan
- Prior art keywords
- immunoglobulin
- amino acid
- binding
- protein
- isolated
- 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
- 102000028557 immunoglobulin binding proteins Human genes 0.000 title claims description 87
- 108091009323 immunoglobulin binding proteins Proteins 0.000 title claims description 87
- 230000027455 binding Effects 0.000 claims description 151
- 108010088160 Staphylococcal Protein A Proteins 0.000 claims description 112
- 108060003951 Immunoglobulin Proteins 0.000 claims description 96
- 102000018358 immunoglobulin Human genes 0.000 claims description 96
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical group NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 90
- 235000004279 alanine Nutrition 0.000 claims description 71
- 108090000623 proteins and genes Proteins 0.000 claims description 64
- 239000004471 Glycine Chemical group 0.000 claims description 58
- 102000004169 proteins and genes Human genes 0.000 claims description 54
- 235000018102 proteins Nutrition 0.000 claims description 52
- 125000003275 alpha amino acid group Chemical group 0.000 claims description 46
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical group C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims description 45
- 235000001014 amino acid Nutrition 0.000 claims description 41
- 125000003630 glycyl group Chemical group [H]N([H])C([H])([H])C(*)=O 0.000 claims description 41
- 229940024606 amino acid Drugs 0.000 claims description 39
- 150000001413 amino acids Chemical class 0.000 claims description 35
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 claims description 28
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 27
- 125000000539 amino acid group Chemical group 0.000 claims description 26
- 229920001184 polypeptide Polymers 0.000 claims description 24
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 24
- 150000007523 nucleic acids Chemical class 0.000 claims description 23
- 238000000034 method Methods 0.000 claims description 19
- 229940072221 immunoglobulins Drugs 0.000 claims description 16
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims description 12
- 239000004472 Lysine Substances 0.000 claims description 12
- 239000004475 Arginine Chemical group 0.000 claims description 10
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical group NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims description 10
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical group NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 claims description 10
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Chemical group CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 claims description 10
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Chemical group OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 10
- 108020004707 nucleic acids Proteins 0.000 claims description 10
- 102000039446 nucleic acids Human genes 0.000 claims description 10
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 claims description 8
- 239000012504 chromatography matrix Substances 0.000 claims description 8
- 239000007787 solid Substances 0.000 claims description 7
- 239000011159 matrix material Substances 0.000 claims description 6
- 108091006020 Fc-tagged proteins Proteins 0.000 claims description 3
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 2
- 125000001909 leucine group Chemical group [H]N(*)C(C(*)=O)C([H])([H])C(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 2
- 125000000613 asparagine group Chemical group N[C@@H](CC(N)=O)C(=O)* 0.000 claims 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 38
- 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 36
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 33
- 238000002474 experimental method Methods 0.000 description 24
- 210000004027 cell Anatomy 0.000 description 23
- 239000011347 resin Substances 0.000 description 22
- 229920005989 resin Polymers 0.000 description 22
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 21
- 239000004473 Threonine Substances 0.000 description 21
- 102220605242 Heparan sulfate N-sulfotransferase 4_G29K_mutation Human genes 0.000 description 20
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 20
- 238000004458 analytical method Methods 0.000 description 20
- 102220149729 rs763469367 Human genes 0.000 description 20
- 102220094399 rs777971423 Human genes 0.000 description 19
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 18
- 239000002953 phosphate buffered saline Substances 0.000 description 18
- 125000003729 nucleotide group Chemical group 0.000 description 17
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 16
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 15
- 108091028043 Nucleic acid sequence Proteins 0.000 description 14
- 239000012634 fragment Substances 0.000 description 14
- 239000002773 nucleotide Substances 0.000 description 13
- 108020004414 DNA Proteins 0.000 description 12
- 239000006228 supernatant Substances 0.000 description 11
- 239000013598 vector Substances 0.000 description 11
- 238000001042 affinity chromatography Methods 0.000 description 9
- 238000003752 polymerase chain reaction Methods 0.000 description 9
- 229910052759 nickel Inorganic materials 0.000 description 8
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 8
- 239000000427 antigen Substances 0.000 description 7
- 102000036639 antigens Human genes 0.000 description 7
- 108091007433 antigens Proteins 0.000 description 7
- 235000009697 arginine Nutrition 0.000 description 7
- 229960003121 arginine Drugs 0.000 description 7
- 235000005772 leucine Nutrition 0.000 description 7
- 229960003136 leucine Drugs 0.000 description 7
- 235000018977 lysine Nutrition 0.000 description 7
- 229960003646 lysine Drugs 0.000 description 7
- 239000013612 plasmid Substances 0.000 description 7
- 108091033319 polynucleotide Proteins 0.000 description 7
- 239000002157 polynucleotide Substances 0.000 description 7
- 102000040430 polynucleotide Human genes 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- 238000012986 modification Methods 0.000 description 6
- 230000004048 modification Effects 0.000 description 6
- 229920000936 Agarose Polymers 0.000 description 5
- 239000012515 MabSelect SuRe Substances 0.000 description 5
- 229960000723 ampicillin Drugs 0.000 description 5
- 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 5
- 239000012491 analyte Substances 0.000 description 5
- 239000000872 buffer Substances 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 238000010494 dissociation reaction Methods 0.000 description 5
- 230000005593 dissociations Effects 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 4
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 238000004587 chromatography analysis Methods 0.000 description 4
- 238000010828 elution Methods 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 235000014304 histidine Nutrition 0.000 description 4
- 229960002885 histidine Drugs 0.000 description 4
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 4
- 238000006467 substitution reaction Methods 0.000 description 4
- 235000000346 sugar Nutrition 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Natural products OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 description 3
- 102000018071 Immunoglobulin Fc Fragments Human genes 0.000 description 3
- 108010091135 Immunoglobulin Fc Fragments Proteins 0.000 description 3
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 3
- 102000008300 Mutant Proteins Human genes 0.000 description 3
- 108010021466 Mutant Proteins Proteins 0.000 description 3
- 235000009582 asparagine Nutrition 0.000 description 3
- 229960001230 asparagine Drugs 0.000 description 3
- 239000005289 controlled pore glass Substances 0.000 description 3
- 239000013604 expression vector Substances 0.000 description 3
- 239000003446 ligand Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 235000006109 methionine Nutrition 0.000 description 3
- 229930182817 methionine Natural products 0.000 description 3
- 229960004452 methionine Drugs 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 108091008146 restriction endonucleases Proteins 0.000 description 3
- 238000010561 standard procedure Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 150000008163 sugars Chemical class 0.000 description 3
- 230000002194 synthesizing effect Effects 0.000 description 3
- 238000001262 western blot Methods 0.000 description 3
- MTCFGRXMJLQNBG-REOHCLBHSA-N (2S)-2-Amino-3-hydroxypropansäure Chemical compound OC[C@H](N)C(O)=O MTCFGRXMJLQNBG-REOHCLBHSA-N 0.000 description 2
- 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 description 2
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 2
- 108020004705 Codon Proteins 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 241001198387 Escherichia coli BL21(DE3) Species 0.000 description 2
- 241000287828 Gallus gallus Species 0.000 description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 2
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 2
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 2
- ONIBWKKTOPOVIA-BYPYZUCNSA-N L-Proline Chemical compound OC(=O)[C@@H]1CCCN1 ONIBWKKTOPOVIA-BYPYZUCNSA-N 0.000 description 2
- DCXYFEDJOCDNAF-REOHCLBHSA-N L-asparagine Chemical compound OC(=O)[C@@H](N)CC(N)=O DCXYFEDJOCDNAF-REOHCLBHSA-N 0.000 description 2
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-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
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-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
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 2
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical group O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 2
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 2
- 150000001412 amines Chemical class 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 235000003704 aspartic acid Nutrition 0.000 description 2
- 229960005261 aspartic acid Drugs 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 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 2
- 238000004113 cell culture Methods 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000012539 chromatography resin Substances 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 235000018417 cysteine Nutrition 0.000 description 2
- 229960002433 cysteine Drugs 0.000 description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 210000003527 eukaryotic cell Anatomy 0.000 description 2
- 108020001507 fusion proteins Proteins 0.000 description 2
- 102000037865 fusion proteins Human genes 0.000 description 2
- 235000013922 glutamic acid Nutrition 0.000 description 2
- 229960002989 glutamic acid Drugs 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- 235000004554 glutamine Nutrition 0.000 description 2
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 2
- 229960002743 glutamine Drugs 0.000 description 2
- 239000011544 gradient gel Substances 0.000 description 2
- 235000014705 isoleucine Nutrition 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
- 125000005647 linker group Chemical group 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 235000008729 phenylalanine Nutrition 0.000 description 2
- 229960005190 phenylalanine Drugs 0.000 description 2
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 2
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 210000001236 prokaryotic cell Anatomy 0.000 description 2
- 235000013930 proline Nutrition 0.000 description 2
- 229960002429 proline Drugs 0.000 description 2
- 235000004400 serine Nutrition 0.000 description 2
- 229960001153 serine Drugs 0.000 description 2
- 125000000430 tryptophan group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C2=C([H])C([H])=C([H])C([H])=C12 0.000 description 2
- 235000002374 tyrosine Nutrition 0.000 description 2
- 229960004441 tyrosine Drugs 0.000 description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 2
- 235000014393 valine Nutrition 0.000 description 2
- 229960004295 valine Drugs 0.000 description 2
- 239000004474 valine Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 1
- QFVHZQCOUORWEI-UHFFFAOYSA-N 4-[(4-anilino-5-sulfonaphthalen-1-yl)diazenyl]-5-hydroxynaphthalene-2,7-disulfonic acid Chemical compound C=12C(O)=CC(S(O)(=O)=O)=CC2=CC(S(O)(=O)=O)=CC=1N=NC(C1=CC=CC(=C11)S(O)(=O)=O)=CC=C1NC1=CC=CC=C1 QFVHZQCOUORWEI-UHFFFAOYSA-N 0.000 description 1
- -1 9-fluorenylmethyloxycarbonyl Chemical group 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 102000014914 Carrier Proteins Human genes 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 108700024394 Exon Proteins 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 108091006054 His-tagged proteins Proteins 0.000 description 1
- 102000017727 Immunoglobulin Variable Region Human genes 0.000 description 1
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 description 1
- 108091092195 Intron Proteins 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- CERQOIWHTDAKMF-UHFFFAOYSA-M Methacrylate Chemical compound CC(=C)C([O-])=O CERQOIWHTDAKMF-UHFFFAOYSA-M 0.000 description 1
- 102100035971 Molybdopterin molybdenumtransferase Human genes 0.000 description 1
- 101710119577 Molybdopterin molybdenumtransferase Proteins 0.000 description 1
- 108020004711 Nucleic Acid Probes Proteins 0.000 description 1
- 108091034117 Oligonucleotide Proteins 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 108091028664 Ribonucleotide Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 101000980867 Schizosaccharomyces pombe (strain 972 / ATCC 24843) Curved DNA-binding protein Proteins 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- 101000982319 Shallot virus X Uncharacterized ORF4 protein Proteins 0.000 description 1
- 108020004682 Single-Stranded DNA Proteins 0.000 description 1
- 101000582398 Staphylococcus aureus Replication initiation protein Proteins 0.000 description 1
- 108091061763 Triple-stranded DNA Proteins 0.000 description 1
- 239000007984 Tris EDTA buffer Substances 0.000 description 1
- 239000007983 Tris buffer Substances 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
- 230000002378 acidificating effect Effects 0.000 description 1
- 238000001261 affinity purification Methods 0.000 description 1
- 150000003862 amino acid derivatives Chemical class 0.000 description 1
- 150000001508 asparagines Chemical group 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 150000001540 azides Chemical class 0.000 description 1
- 108091008324 binding proteins Proteins 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- 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 1
- 239000012501 chromatography medium Substances 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 239000005547 deoxyribonucleotide Substances 0.000 description 1
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 150000002333 glycines Chemical group 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
- 238000011534 incubation Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 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 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- 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 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 239000012516 mab select resin Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 238000001823 molecular biology technique Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 108091005763 multidomain proteins Proteins 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000219 mutagenic Toxicity 0.000 description 1
- 230000003505 mutagenic effect Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 230000002352 nonmutagenic effect Effects 0.000 description 1
- 239000002853 nucleic acid probe Substances 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000004091 panning Methods 0.000 description 1
- 238000010647 peptide synthesis reaction Methods 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000002731 protein assay Methods 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 239000002336 ribonucleotide Substances 0.000 description 1
- 125000002652 ribonucleotide group Chemical group 0.000 description 1
- 102220032940 rs367543239 Human genes 0.000 description 1
- 102220285342 rs777971423 Human genes 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001488 sodium phosphate Substances 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000012064 sodium phosphate buffer Substances 0.000 description 1
- 239000007790 solid phase Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000000527 sonication Methods 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 125000005931 tert-butyloxycarbonyl group Chemical group [H]C([H])([H])C(OC(*)=O)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 125000000341 threoninyl group Chemical group [H]OC([H])(C([H])([H])[H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 241001515965 unidentified phage Species 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/12—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria
- C07K16/1267—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-positive bacteria
- C07K16/1271—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from bacteria from Gram-positive bacteria from Micrococcaceae (F), e.g. Staphylococcus
-
- 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
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/281—Sorbents specially adapted for preparative, analytical or investigative chromatography
- B01J20/286—Phases chemically bonded to a substrate, e.g. to silica or to polymers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/32—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating
- B01J20/3231—Impregnating or coating ; Solid sorbent compositions obtained from processes involving impregnating or coating characterised by the coating or impregnating layer
- B01J20/3242—Layers with a functional group, e.g. an affinity material, a ligand, a reactant or a complexing group
- B01J20/3268—Macromolecular compounds
- B01J20/3272—Polymers obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
- B01J20/3274—Proteins, nucleic acids, polysaccharides, antibodies or antigens
-
- 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
- C07K14/305—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F)
- C07K14/31—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Micrococcaceae (F) from Staphylococcus (G)
Description
本願は、2008年8月11日に出願された米国仮特許出願61/188,549号および2009年4月16日に出願された米国仮特許出願61/212,812号(これらの各々の内容全体が、参照により、本明細書中に組み込まれる。)の優先権の利益を主張する。
本発明は、免疫グロブリンに対して改善された結合特異性を有する、修飾された免疫グロブリン結合タンパク質、例えば、ブドウ球菌プロテインA、並びにこれを製造及び使用する方法に関する。
本開示をより容易に理解できるようにするために、まず、ある種の用語を定義する。さらなる定義は、詳細な説明を通じて記載する。
SpAは、細菌スタフィロコッカス・オーレウスに由来する約42kDaのタンパク質であり、N末端に5つの直列の高度に相同な細胞外免疫グロブリン(Ig)結合ドメイン(E、D、A、B及びCと表記される。)を含有する。SpAの各細胞外ドメインは、異なるIg結合部位を有する。1つの部位はFcγ(IgのIgGクラスの定常領域)に対するものであり、他の部位は、ある種のIg分子のFab部分(抗原認識に必要なIgの部分)に対するものである。ドメインの各々がFab結合部位を含有すると報告されている。SpAの非Ig結合部分はC末端に位置しており、X領域又はXドメインと表記される。
本発明のSpA変異物は、本分野において公知のあらゆる適切な方法を用いて作製することができる。例えば、Sambrook、Fritsch及びManiatisによるMolecular Cloningという題名の研究室マニュアル中に記載されているものなどの、核酸の部位特異的突然変異誘発のための標準的な技術を例えば使用し得る。さらに、ポリメラーゼ連鎖反応(PCR)突然変異誘発を含む標準的な分子生物学技術を使用し得る。
SpA変異物の作製に続いて、免疫グロブリンのFab又はFc部分へのSpA変異物の結合特異性は、本分野における標準的な技術及び本明細書中に記載されている技術を用いて決定される。
本発明は、抗体のFc部分に対する親和性を有し、及び抗体のFab部分に対して低下した親和性を有する又は親和性を有さない少なくとも1つの免疫グロブリン結合タンパク質を含むクロマトグラフィーマトリックスを調製する方法も提供する。一実施形態において、このような方法は、(a)単離されたSpAドメイン(例えば、E、D、A、B、C又はZ)をコードする核酸配列を準備すること;(b)アラニン又はトリプトファン以外のアミノ酸によって、少なくとも29位のグリシンが置換されている変異物タンパク質をコードするように前記核酸配列を変異させること;(c)宿主細胞(例えば、適切な原核細胞又は真核細胞)中の変異物タンパク質を発現させること;(d)変異物タンパク質を宿主細胞から回収すること;及び(e)固体支持体へ変異物タンパク質を連結させることを含む。
SpA変異物を含有するベクターの構築
プロテインAの「Bドメイン」をコードする遺伝子は、DNA合成装置を用いた標準的な方法によって合成される。遺伝子の5’末端は、開始メチオニンに対するコドンを含み、及び遺伝子の3’末端に6つのヒスチジンコドンを含む。この遺伝子は、ベクターpJ56:8620中に与えられる。親ベクターpJ56は、アンピシリン、カナマイシン、クロラムフェニコール及びゲンタマイシンに対する耐性を付与する。続いて発現ベクター中に遺伝子をクローニングするための適切な制限酵素部位が、遺伝子の5’末端及び3’末端の両方に導入される。ベクターのプラスミド地図が図5に示されている。
プロテインA変異物の発現及び精製
様々なSpA変異物を発現させるために、あらゆる適切な細菌発現系を使用することができる。例えば、タンパク質は、pET11a(Novagen,MadisonWI)などのpETベクターから、株BL21(DE3)(Promega,MadisonWI)などのエシェリヒア・コリ(Eschericia coli)中で発現され得る。プレートから単一コロニーを拾い上げ、100μg/mLアンピシリンを含有するLB培地中において、約37℃で一晩増殖させる。100μg/mLアンピシリンを含有する新鮮なLB培地中に、一晩培養物を100倍希釈し、600nmの光学密度が0.8であるような細胞密度まで増殖させる。1mMイソプロピル−β−D−チオガラクトピラノシドの添加後、細胞をさらに2時間増殖させる。SDS−PAGE分析及びウェスタンブロッティングによって発現を確認する。典型的なプロテインAウェスタンブロットが図8に示されており、図8は、イー・コリBL21(DE3)細胞中のベクターPET11a:8620からのプロテインAの発現がニワトリIgY抗プロテインA抗体を用いて検出されることを示している。
Fc及びFab断片へのSpA変異物の結合
続いて、NiNTAアガロース樹脂(Qiagen)を用いて、Fc及びFabを結合する能力に関して、各SpA変異物を試験する。JacksonImmunoResearch(WestGrove,PA)から、Fc及びF(ab)2断片を入手する。20mMイミダゾールを含有するリン酸緩衝化生理的食塩水中に、0.11mg/mLの濃度まで各SpA変異物を希釈し、0.44mg/mLの濃度までFcを希釈し、0.92mg/mLの濃度までF(ab)2を希釈する。NiNTAアガロース樹脂を再懸濁し、樹脂スラリー約20μLを、0.5mLの微小遠心管中にピペットで分注する。続いて、樹脂を沈降させ、上清を廃棄する。20mMイミダゾールを加えたPBS約100μLで樹脂を洗浄し、室温で約15分間温置し、再度沈降させる。上清を廃棄する。約250μLの0.11mg/mLの各SpA変異物又はPBS対照を、2つ組みで、樹脂カラム上に搭載し、室温で約30分間温置する。樹脂を沈降させ、分析のために上清を保存する。毎回、20mMイミダゾールを加えたPBS100μLで、沈降物を3回洗浄し、5分間温置し、再度沈降させる。上清を廃棄する。樹脂とともに約30分間温置することによって、SpA変異物の各々又はPBS対照との結合に関して、Fc又はF(ab)2の約100μLを試験する。続いて、樹脂を沈降させ、分析のために上清を保存する。毎回、20mMイミダゾールを加えたPBS約100μLで、樹脂を再度3回洗浄し、5分間温置し、沈降させる。今回は、上清を廃棄する。20mMイミダゾールを加えたPBS約100μLで、結合された画分を溶出し、約15分間温置し、沈降させる。さらなる分析のために、上清を保存する。続いて、図9から12に図示されているように、Bio−RadLaboratories,CAから購入した4から15%勾配のゲル上でのSDS−PAGEによって、試料を分析する。
Fab binding: Melissa A. Starovasnik, Mark P. O’connell, Wayne J. Fairbrother, And Robert F. Kelley (1999). Antibody variable region binding by Staphylococal proteinA: Thermodynamic analysis and location of the Fv binding site on E-domain. Protein Science 8, 1423-1431.
Fc binding constant: Susanne Gulich, Mathias Uhlen, Sophia Hober (2000) Protein engineering of an IgG-binding domain allows milder elution conditions during affinity chromatography. Journal of Biotechnology 76, 233-244
Claims (16)
- ブドウ球菌プロテインAの1つ又はそれ以上の単離されたドメインを含む単離された免疫グロブリン結合タンパク質であって、前記1つ又はそれ以上の単離されたドメインは、配列番号10、配列番号11又は配列番号12に記載されているアミノ酸配列、もしくは配列番号10、11又は12に少なくとも90%同一である配列を含み、及び前記1つ又はそれ以上の単離されたドメインは、(i)前記ドメインが配列番号10又は11に記載されているアミノ酸配列、もしくは配列番号10又は11に少なくとも90%同一性を有するアミノ酸配列を含む場合には、少なくとも29位のグリシン残基がリジン、アルギニン及びロイシンからなる群から選択されるアミノ酸残基で置換されており、或いは(ii)前記ドメインが配列番号12に記載されているアミノ酸配列、もしくは配列番号12に少なくとも90%同一性を有するアミノ酸配列を含む場合には、少なくとも29位のアラニン残基がリジン、アルギニン及びロイシンからなる群から選択されるアミノ酸残基で置換されており、
前記免疫グロブリン結合タンパク質は、免疫グロブリンのFc部分を結合するが、29位にアラニン又はグリシンを含む免疫グロブリン結合タンパク質と比べて、免疫グロブリンのFab部分への低下した結合を示す、前記単離された免疫グロブリン結合タンパク質。 - ブドウ球菌プロテインAのカルボキシ末端領域の少なくとも一部をさらに含む、請求項1に記載の単離された免疫グロブリン結合タンパク質。
- 固体支持体へ連結された、請求項1に記載の単離された免疫グロブリン結合タンパク質を含むクロマトグラフィーマトリックス。
- 請求項1に記載の免疫グロブリン結合タンパク質を含むポリペプチドのライブラリー。
- 配列番号10又は配列番号12は、さらに23位のアスパラギン残基が別のアミノ酸で置換されている、請求項1に記載の免疫グロブリン結合タンパク質。
- 免疫グロブリンがIgGである、請求項1に記載の免疫グロブリン結合タンパク質。
- 請求項1に記載の免疫グロブリン結合タンパク質をコードする核酸分子。
- 請求項7に記載の核酸分子を含む宿主細胞。
- (a)1つ又はそれ以上の免疫グロブリンを含む試料を準備する工程;
(b)前記1つ又はそれ以上の免疫グロブリンがマトリックスに結合するような条件下で、請求項3に記載のマトリックスと試料を接触する工程;及び
(c)適切な条件下で溶出することによって、1つ又はそれ以上の結合された免疫グロブリンを回収する工程;
を含む、試料から1つ又はそれ以上の免疫グロブリンをアフィニティー精製する方法。 - 固体支持体へ連結された、請求項2に記載の単離された免疫グロブリン結合タンパク質を含むクロマトグラフィーマトリックス。
- (a)1つ又はそれ以上の免疫グロブリンを含む試料を準備する工程;
(b)前記1つ又はそれ以上の免疫グロブリンがマトリックスに結合するような条件下で、請求項10に記載のマトリックスと試料を接触する工程;及び
(c)適切な条件下で溶出することによって、1つ又はそれ以上の結合された免疫グロブリンを回収する工程;
を含む、試料から1つ又はそれ以上の免疫グロブリンをアフィニティー精製する方法。 - 免疫グロブリンがIgG、IgA及びIgMからなる群から選択される、請求項2に記載の免疫グロブリン結合タンパク質。
- 免疫グロブリンのFc部分がFc融合タンパク質の一部である、請求項1に記載の免疫グロブリン結合タンパク質。
- ブドウ球菌プロテインAの1つ又はそれ以上の単離されたドメインを含む単離された免疫グロブリン結合タンパク質であって、前記1つ又はそれ以上の単離されたドメインは、配列番号10又は12に記載されているアミノ酸配列を含み、ここで配列番号10の場合には少なくとも29位のグリシン残基が、或いは配列番号12の場合には少なくとも29位のアラニン残基が、リジン、ロイシン及びアルギニンからなる群から選択されるアミノ酸残基で置換されており、
前記免疫グロブリン結合タンパク質は、免疫グロブリンのFc部分を結合するが、野生型プロテインAと比べて免疫グロブリンのFab部分への低下した結合を示す、前記単離された免疫グロブリン結合タンパク質。 - ブドウ球菌プロテインAの1つ又はそれ以上の単離されたCドメインを含む単離された免疫グロブリン結合タンパク質であって、前記1つ又はそれ以上の単離されたCドメインは、配列番号11に記載されているアミノ酸配列、もしくは配列番号11のアミノ酸配列に少なくとも90%同一性を有するアミノ酸配列を含み、ここで少なくとも29位のグリシン残基がリジンアミノ酸残基で置換されており、
前記免疫グロブリン結合タンパク質は、免疫グロブリンのFc部分を結合するが、野生型プロテインAと比べて免疫グロブリンのFab部分への低下した結合を示す、前記単離された免疫グロブリン結合タンパク質。 - ブドウ球菌プロテインAの1つ又はそれ以上の単離されたCドメインを含む単離された免疫グロブリン結合タンパク質であって、前記1つ又はそれ以上の単離されたCドメインは、少なくとも29位のグリシン残基がリジンアミノ酸残基で置換されている、前記単離された免疫グロブリン結合タンパク質。
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US18854908P | 2008-08-11 | 2008-08-11 | |
US61/188,549 | 2008-08-11 | ||
US21281209P | 2009-04-16 | 2009-04-16 | |
US61/212,812 | 2009-04-16 |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2013117518A Division JP5913202B2 (ja) | 2008-08-11 | 2013-06-04 | 改善された特異性を有する新規免疫グロブリン結合タンパク質 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2010075175A JP2010075175A (ja) | 2010-04-08 |
JP5480558B2 true JP5480558B2 (ja) | 2014-04-23 |
Family
ID=41258393
Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2009185389A Active JP5480558B2 (ja) | 2008-08-11 | 2009-08-10 | 改善された特異性を有する新規免疫グロブリン結合タンパク質 |
JP2013117518A Active JP5913202B2 (ja) | 2008-08-11 | 2013-06-04 | 改善された特異性を有する新規免疫グロブリン結合タンパク質 |
JP2015254325A Active JP6181145B2 (ja) | 2008-08-11 | 2015-12-25 | 改善された特異性を有する新規免疫グロブリン結合タンパク質 |
Family Applications After (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2013117518A Active JP5913202B2 (ja) | 2008-08-11 | 2013-06-04 | 改善された特異性を有する新規免疫グロブリン結合タンパク質 |
JP2015254325A Active JP6181145B2 (ja) | 2008-08-11 | 2015-12-25 | 改善された特異性を有する新規免疫グロブリン結合タンパク質 |
Country Status (7)
Country | Link |
---|---|
US (2) | US8592555B2 (ja) |
EP (3) | EP2495253B1 (ja) |
JP (3) | JP5480558B2 (ja) |
CN (3) | CN104804073B (ja) |
ES (3) | ES2612138T3 (ja) |
IN (1) | IN2015DE00245A (ja) |
SG (2) | SG193852A1 (ja) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPWO2010110288A1 (ja) * | 2009-03-24 | 2012-09-27 | 株式会社カネカ | 免疫グロブリンに親和性を有するタンパク質および免疫グロブリン結合性アフィニティーリガンド |
JP2013226148A (ja) * | 2008-08-11 | 2013-11-07 | Emd Millipore Corp | 改善された特異性を有する新規免疫グロブリン結合タンパク質 |
US9376474B1 (en) | 2011-06-08 | 2016-06-28 | Emd Millipore Corporation | Chromatography matrices including novel Staphylococcus aureus protein a based ligands |
US10072050B2 (en) | 2008-12-24 | 2018-09-11 | Emd Millipore Corporation | Caustic stable chromatography ligands |
Families Citing this family (31)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20120149875A1 (en) * | 2009-01-12 | 2012-06-14 | Ge Healthcare Bio-Sciences Ab | Affinity chromatography matrix |
HUE026855T2 (en) | 2009-04-03 | 2016-07-28 | Univ Chicago | Preparations and method for Protein A (SPA) variants |
AU2011222991B2 (en) * | 2010-03-05 | 2015-07-16 | Boehringer Ingelheim International Gmbh | Selective enrichment of antibodies |
JP6002128B2 (ja) | 2010-07-02 | 2016-10-05 | ザ・ユニバーシティ・オブ・シカゴThe University Of Chicago | プロテインA(SpA)変種に関連する組成物および方法 |
EP2632948B1 (en) | 2010-10-27 | 2016-11-16 | Spiber Technologies AB | Spider silk fusion protein structures for binding to an organic target |
EP2744517B1 (en) * | 2011-08-15 | 2019-03-13 | The University of Chicago | Compositions and methods related to antibodies to staphylococcal protein a |
US9657055B2 (en) * | 2011-11-30 | 2017-05-23 | Ge Healthcare Bioprocess R&D Ab | Affinity chromatography matrix |
CN103014880B (zh) * | 2012-12-20 | 2015-06-24 | 天津大学 | 基于蛋白a亲和模型构建免疫球蛋白g的亲和配基多肽库及设计方法的应用 |
KR20150011231A (ko) * | 2013-07-22 | 2015-01-30 | 삼성디스플레이 주식회사 | 유기 발광 표시 장치 및 이의 제조 방법 |
NO3023777T3 (ja) * | 2013-09-09 | 2018-04-14 | ||
KR102393642B1 (ko) | 2014-01-17 | 2022-05-03 | 리플리겐 코포레이션 | 크로마토그래피 컬럼 살균 |
CN105198973B (zh) * | 2014-06-18 | 2019-01-25 | 上海交通大学 | 一种葡萄球菌a蛋白z结构域衍生物及其应用 |
US11566082B2 (en) | 2014-11-17 | 2023-01-31 | Cytiva Bioprocess R&D Ab | Mutated immunoglobulin-binding polypeptides |
TWI709571B (zh) * | 2015-03-26 | 2020-11-11 | 日商Jsr股份有限公司 | 免疫球蛋白結合蛋白質及使用此之親和性載體 |
US10766924B2 (en) * | 2015-07-28 | 2020-09-08 | Jsr Corporation | Affinity support and method for isolating immunoglobulin |
US11708390B2 (en) | 2016-05-11 | 2023-07-25 | Cytiva Bioprocess R&D Ab | Method of storing a separation matrix |
US10730908B2 (en) | 2016-05-11 | 2020-08-04 | Ge Healthcare Bioprocess R&D Ab | Separation method |
US10513537B2 (en) | 2016-05-11 | 2019-12-24 | Ge Healthcare Bioprocess R&D Ab | Separation matrix |
US10889615B2 (en) | 2016-05-11 | 2021-01-12 | Cytiva Bioprocess R&D Ab | Mutated immunoglobulin-binding polypeptides |
ES2909833T3 (es) | 2016-05-11 | 2022-05-10 | Cytiva Bioprocess R & D Ab | Método de limpieza y/o desinfección de una matriz de separación |
US10703774B2 (en) | 2016-09-30 | 2020-07-07 | Ge Healthcare Bioprocess R&D Ab | Separation method |
JP7181612B2 (ja) * | 2016-08-11 | 2022-12-01 | ナフィゴ プロテインズ ゲゼルシャフト ミット ベシュレンクテル ハフツング | アルカリ安定性免疫グロブリン結合タンパク質 |
EP3665189A1 (en) | 2017-08-07 | 2020-06-17 | Repligen Corporation | Fc binding proteins with cysteine in the c-terminal helical region |
EP3521304B9 (en) * | 2018-02-01 | 2023-10-04 | Repligen Corporation | Fc binding proteins with cysteine in the c-terminal helical region |
BR112021005907A2 (pt) | 2018-09-27 | 2021-08-10 | Xilio Development, Inc. | citocinas mascaradas, ácido nucleico, vetor, célula hospedeira, métodos para produzir uma citocina mascarada, para tratar ou prevenir uma doença neoplásica e para tratar ou prevenir uma doença inflamatória ou autoimune neoplásica, composição, composição farmacêutica e kit |
CN113924320A (zh) * | 2019-04-02 | 2022-01-11 | 邦德威尔科技有限合伙公司 | 用于增强抗体纯化的功能化ubx蛋白材料 |
CN111057153B (zh) * | 2019-12-06 | 2021-09-07 | 广州康盛生物科技股份有限公司 | 一种免疫球蛋白结合蛋白及其制备方法和应用 |
CN110950969B (zh) * | 2019-12-23 | 2021-03-30 | 北京全式金生物技术有限公司 | 一种具有免疫球蛋白结合能力的融合蛋白 |
GB202113626D0 (en) | 2021-09-24 | 2021-11-10 | Cytiva Bioprocess R & D Ab | FC binding polypeptides |
CN114409801B (zh) * | 2021-12-22 | 2023-04-18 | 苏州赛分科技股份有限公司 | 基于重组蛋白a的亲和层析介质及其制备方法和应用 |
GB202203478D0 (en) | 2022-03-14 | 2022-04-27 | Cytiva Bioprocess R & D Ab | Vh3 binding polypeptides |
Family Cites Families (66)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
SE8204810L (sv) | 1982-08-23 | 1984-02-24 | Pharmacia Fine Chemicals Ab | Hybrid-dna-molekyl, transformerad mikroorganism och sett att framstella protein a |
US5151350A (en) | 1982-10-27 | 1992-09-29 | Repligen Corporation | Cloned genes encoding recombinant protein a |
US4617266A (en) | 1983-04-28 | 1986-10-14 | Genex Corporation | Production of Protein A |
SE8505922D0 (sv) | 1985-12-13 | 1985-12-13 | Kabigen Ab | Construction of an igg binding protein to facilitate downstream processing using protein engineering |
US5260373A (en) | 1987-03-13 | 1993-11-09 | Repligen Corporation | Immobilized immunoglobulin-binding proteins |
US5084559A (en) | 1987-03-27 | 1992-01-28 | Repligen Corporation | Protein a domain mutants |
US5089473A (en) | 1988-08-29 | 1992-02-18 | Monsanto Company | Somatotropin variants and their use |
GB8902770D0 (en) | 1989-02-08 | 1989-03-30 | Bioprocessing Ltd | Improvements in or relating to supports for immunoaffinity separations |
US4879378A (en) | 1989-03-30 | 1989-11-07 | Union Carbide Chemicals And Plastics Company Inc. | Polysiloxanes with pendant sterically hindered phenol moiety connected to the silicon atom via a carbonylyoxy-containing link |
US5401650A (en) | 1990-10-24 | 1995-03-28 | The Mount Sinai School Of Medicine Of The City University Of New York | Cloning and expression of biologically active α-galactosidase A |
US5198531A (en) | 1991-06-14 | 1993-03-30 | Research Diagnostic Antibodies | Polymeric resin for peptide synthesis |
EP0550771B1 (en) | 1991-07-25 | 1999-09-29 | Oriental Yeast Co., Ltd. | Immunoglobulin-binding artificial protein |
US5240680A (en) | 1991-12-19 | 1993-08-31 | Chiron Corporation | Automated apparatus for use in peptide synthesis |
US5573905A (en) | 1992-03-30 | 1996-11-12 | The Scripps Research Institute | Encoded combinatorial chemical libraries |
SE9400088D0 (sv) | 1994-01-14 | 1994-01-14 | Kabi Pharmacia Ab | Bacterial receptor structures |
SE9503925D0 (sv) | 1995-11-07 | 1995-11-07 | Pharmacia Biotech Ab | Separationsmedium för IgG |
AU2429397A (en) | 1996-03-29 | 1997-10-22 | David S. Terman | Polymerized staphylococcal protein a for treatment of diseases |
US6013763A (en) | 1996-06-04 | 2000-01-11 | Genentech, Inc. | Peptide variants of protein A |
US5856452A (en) | 1996-12-16 | 1999-01-05 | Sembiosys Genetics Inc. | Oil bodies and associated proteins as affinity matrices |
CA2315944A1 (en) | 1997-12-24 | 1999-07-08 | Diatech Pty. Ltd. | Bifunctional molecules |
GB9823071D0 (en) | 1998-10-21 | 1998-12-16 | Affibody Technology Ab | A method |
US6589529B1 (en) | 1998-10-30 | 2003-07-08 | Children's Hospital Medical Center | Rotavirus subunit vaccine |
US6602977B1 (en) | 1999-04-19 | 2003-08-05 | Biovitrum Ab | Receptor structures |
SE9901379D0 (sv) | 1999-04-19 | 1999-04-19 | Pharmacia & Upjohn Ab | Receptor structures |
US7163686B1 (en) | 1999-05-15 | 2007-01-16 | The Regents Of The University Of California | Protein A based binding domains with desirable activities |
AU777005B2 (en) * | 1999-05-15 | 2004-09-30 | University Of California, San Diego | Protein A based binding domains with desirable activities |
WO2003057881A1 (fr) | 2001-12-28 | 2003-07-17 | Chugai Seiyaku Kabushiki Kaisha | Procede de stabilisation d'une proteine |
SE0200943D0 (sv) * | 2002-03-25 | 2002-03-25 | Amersham Biosciences Ab | Mutant protein |
US7709209B2 (en) | 2002-03-25 | 2010-05-04 | Ge Healthcare Bio-Sciences Ab | Protein ligands |
US7083948B1 (en) | 2002-12-24 | 2006-08-01 | Immunex Corporation | Polypeptide purification reagents and methods for their use |
GB0304576D0 (en) | 2003-02-28 | 2003-04-02 | Lonza Biologics Plc | Protein a chromatography |
DK1601697T3 (da) | 2003-02-28 | 2007-10-01 | Lonza Biologics Plc | Oprensning af antistof ved protein A- og ionbytningskromatografi |
AU2004253835B2 (en) | 2003-07-04 | 2009-01-29 | Affibody Ab | Polypeptides having binding affinity for HER2 |
US7956165B2 (en) | 2003-07-24 | 2011-06-07 | Affisink Biotechnology Ltd. | Compositions and methods for purifying and crystallizing molecules of interest |
US7192738B2 (en) | 2003-10-03 | 2007-03-20 | Genentech, Inc. | IGF binding proteins |
EP1564286A1 (en) | 2004-02-11 | 2005-08-17 | Université de Liège | Hybrid proteins of beta-lactamase class A |
WO2006004067A1 (ja) | 2004-07-06 | 2006-01-12 | Kaneka Corporation | ブレビバチルス属細菌を用いたプロテインa様蛋白質の生産方法 |
US8728828B2 (en) | 2004-12-22 | 2014-05-20 | Ge Healthcare Bio-Sciences Ab | Purification of immunoglobulins |
US20080254551A1 (en) | 2004-12-30 | 2008-10-16 | Achsel Tilmann | Immunoprecipitaion-Based Method to Purify and Characterise Biological Macromolecular Complexes |
WO2006092338A2 (en) | 2005-03-01 | 2006-09-08 | Affibody Ab | Tnf-alpha binding polypeptide , uses thereof and methods employing it |
JP2006304633A (ja) | 2005-04-26 | 2006-11-09 | Apro Life Science Institute Inc | イムノグロブリン結合タンパク質 |
WO2006138553A2 (en) | 2005-06-17 | 2006-12-28 | Wyeth | Methods of purifying fc region containing proteins |
WO2007019376A2 (en) | 2005-08-03 | 2007-02-15 | Rq Bioscience, Inc. | Methods and compositions for diagnosis of iga-and igm-mediated kidney diseases |
CN102417557B (zh) | 2005-08-03 | 2015-03-11 | 默克专利股份公司 | 亲水交联聚合物 |
US8440191B2 (en) | 2005-11-18 | 2013-05-14 | Tufts Medical Center | Clearance of abnormal IGA1 in IGA1 deposition diseases |
US7833723B2 (en) | 2006-01-06 | 2010-11-16 | Millipore Corporation | Affinity chromatography matrices and methods of making and using the same |
EP1992692B1 (en) | 2006-02-21 | 2013-01-09 | Protenova Co., Ltd. | Immunoglobulin affinity ligand |
US20080096819A1 (en) | 2006-05-02 | 2008-04-24 | Allozyne, Inc. | Amino acid substituted molecules |
GB0610776D0 (en) | 2006-05-31 | 2006-07-12 | Univ Bath | Novel applications for staphylococcus aureus Sbi protein |
CN106422418B (zh) | 2006-09-29 | 2022-03-01 | 思拓凡生物工艺研发有限公司 | 用于抗体分离的包含来自金黄色葡萄球菌a蛋白的结构域c的层析配体 |
JP5004165B2 (ja) | 2006-10-10 | 2012-08-22 | 独立行政法人産業技術総合研究所 | タンパク質の配向制御固定化に適したタンパク質 |
US7691608B2 (en) | 2006-12-06 | 2010-04-06 | Repligen Corporation | Nucleic acids encoding recombinant protein A |
US8362217B2 (en) | 2006-12-21 | 2013-01-29 | Emd Millipore Corporation | Purification of proteins |
WO2008091740A2 (en) | 2007-01-22 | 2008-07-31 | Genentech, Inc. | Polyelectrolyte precipitation and purification of antibodies |
JP2008255046A (ja) | 2007-04-04 | 2008-10-23 | Toray Ind Inc | 融合タンパク質、これを用いた抗体捕捉担体および抗原検出法 |
JP2008266221A (ja) | 2007-04-20 | 2008-11-06 | National Institute Of Advanced Industrial & Technology | アミノ末端1箇所で配向制御固定化された固定化タンパク質 |
US8198409B2 (en) | 2007-05-21 | 2012-06-12 | Nomadic Bioscience Co., Ltd. | Polypeptide, an affinity chromatography material, and a method for separating and/or purifying immunoglobulin |
SG149759A1 (en) | 2007-07-10 | 2009-02-27 | Millipore Corp | Media for affinity chromatography |
CN103396480A (zh) | 2008-05-15 | 2013-11-20 | 诺沃—诺迪斯克有限公司 | 抗体纯化方法 |
JP5677943B2 (ja) | 2008-06-05 | 2015-02-25 | アフィボディ・アーベーAffibody Ab | ポリペプチド |
US8592555B2 (en) * | 2008-08-11 | 2013-11-26 | Emd Millipore Corporation | Immunoglobulin-binding proteins with improved specificity |
SG195555A1 (en) | 2008-12-24 | 2013-12-30 | Emd Millipore Corp | Caustic stable chromatography ligands |
US20120149875A1 (en) | 2009-01-12 | 2012-06-14 | Ge Healthcare Bio-Sciences Ab | Affinity chromatography matrix |
EP2412809B1 (en) | 2009-03-24 | 2017-08-09 | Kaneka Corporation | Protein having affinity for immunoglobulin, and immunoglobulin-binding affinity ligand |
EP2646126A4 (en) | 2010-11-29 | 2014-12-17 | Ge Healthcare Bio Sciences Ab | AFFINITY CHROMATOGRAPHY MATRIX |
SG10201604559TA (en) | 2011-06-08 | 2016-07-28 | Emd Millipore Corp | Chromatography matrices including novel staphylococcus aureus protein a based ligands |
-
2009
- 2009-08-04 US US12/462,484 patent/US8592555B2/en active Active
- 2009-08-07 SG SG2013066568A patent/SG193852A1/en unknown
- 2009-08-07 SG SG200905295-2A patent/SG159458A1/en unknown
- 2009-08-10 JP JP2009185389A patent/JP5480558B2/ja active Active
- 2009-08-11 EP EP12163614.6A patent/EP2495253B1/en active Active
- 2009-08-11 EP EP09167670.0A patent/EP2157099B1/en active Active
- 2009-08-11 CN CN201510113971.1A patent/CN104804073B/zh active Active
- 2009-08-11 ES ES12163614.6T patent/ES2612138T3/es active Active
- 2009-08-11 EP EP12163615.3A patent/EP2495254B1/en active Active
- 2009-08-11 ES ES09167670.0T patent/ES2612114T3/es active Active
- 2009-08-11 CN CN200910221476.7A patent/CN101704879B/zh active Active
- 2009-08-11 ES ES12163615.3T patent/ES2612166T3/es active Active
- 2009-08-11 CN CN201410389062.6A patent/CN104163866A/zh active Pending
-
2013
- 2013-06-04 JP JP2013117518A patent/JP5913202B2/ja active Active
- 2013-10-23 US US14/061,080 patent/US9920112B2/en active Active
-
2015
- 2015-01-28 IN IN245DE2015 patent/IN2015DE00245A/en unknown
- 2015-12-25 JP JP2015254325A patent/JP6181145B2/ja active Active
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2013226148A (ja) * | 2008-08-11 | 2013-11-07 | Emd Millipore Corp | 改善された特異性を有する新規免疫グロブリン結合タンパク質 |
JP2016121152A (ja) * | 2008-08-11 | 2016-07-07 | イー・エム・デイー・ミリポア・コーポレイシヨン | 改善された特異性を有する新規免疫グロブリン結合タンパク質 |
US9920112B2 (en) | 2008-08-11 | 2018-03-20 | Emd Millipore Corporation | Immunoglobulin-binding proteins with improved specificity |
US10072050B2 (en) | 2008-12-24 | 2018-09-11 | Emd Millipore Corporation | Caustic stable chromatography ligands |
US11084851B2 (en) | 2008-12-24 | 2021-08-10 | Emd Millipore Corporation | Caustic stable chromatography ligands |
JPWO2010110288A1 (ja) * | 2009-03-24 | 2012-09-27 | 株式会社カネカ | 免疫グロブリンに親和性を有するタンパク質および免疫グロブリン結合性アフィニティーリガンド |
US9376474B1 (en) | 2011-06-08 | 2016-06-28 | Emd Millipore Corporation | Chromatography matrices including novel Staphylococcus aureus protein a based ligands |
Also Published As
Publication number | Publication date |
---|---|
ES2612114T3 (es) | 2017-05-12 |
JP2016121152A (ja) | 2016-07-07 |
JP2010075175A (ja) | 2010-04-08 |
CN101704879B (zh) | 2015-04-08 |
SG159458A1 (en) | 2010-03-30 |
EP2157099B1 (en) | 2016-12-21 |
EP2495253B1 (en) | 2016-12-21 |
EP2495254B1 (en) | 2016-12-21 |
IN2015DE00245A (ja) | 2015-07-17 |
JP6181145B2 (ja) | 2017-08-16 |
US20140046037A1 (en) | 2014-02-13 |
CN101704879A (zh) | 2010-05-12 |
JP5913202B2 (ja) | 2016-04-27 |
US9920112B2 (en) | 2018-03-20 |
US20100063256A1 (en) | 2010-03-11 |
EP2495253A1 (en) | 2012-09-05 |
CN104804073B (zh) | 2018-11-27 |
EP2157099A1 (en) | 2010-02-24 |
ES2612138T3 (es) | 2017-05-12 |
CN104163866A (zh) | 2014-11-26 |
JP2013226148A (ja) | 2013-11-07 |
ES2612166T3 (es) | 2017-05-12 |
EP2495254A1 (en) | 2012-09-05 |
US8592555B2 (en) | 2013-11-26 |
CN104804073A (zh) | 2015-07-29 |
SG193852A1 (en) | 2013-10-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6181145B2 (ja) | 改善された特異性を有する新規免疫グロブリン結合タンパク質 | |
JP7110287B2 (ja) | 腐食安定的クロマトグラフィーリガンド | |
KR102573370B1 (ko) | 신규한 알칼리 안정성 면역글로불린 결합 단백질 | |
JP5933526B2 (ja) | 免疫グロブリン結合性新規ポリペプチド | |
JP6506691B2 (ja) | Fab領域結合性ペプチド | |
KR20180031012A (ko) | 신규한 면역글로불린-결합 단백질 및 친화도 정제에서의 이의 용도 | |
JP2006304633A (ja) | イムノグロブリン結合タンパク質 | |
WO2014021240A1 (ja) | プロテインgの細胞膜外ドメイン変異体のタンデム型多量体から成るタンパク質を含む捕捉剤 | |
AU2020213921B2 (en) | Immunoglobulin binding proteins for affinity purification | |
WO2013018880A1 (ja) | プロテインgの細胞膜外ドメイン変異体のタンデム型多量体から成る新規な改変型タンパク質 | |
JP2019524088A (ja) | アルカリ耐性が向上した変異型免疫グロブリン結合タンパク質 | |
JP2009195184A (ja) | IgG−Fab断片抗体結合性ペプチド | |
WO2015050153A1 (ja) | 免疫グロブリンG(IgG)のFc部分を有するタンパク質に対するアフィニティを有する複数のドメイン間をリンカーで結合させて成るタンパク質 | |
JP2015019615A (ja) | プロテインgの細胞膜外ドメイン変異体の新規な改変型タンパク質 | |
JP2007244285A (ja) | 抗体結合性ペプチド | |
US20220251147A1 (en) | Functionalized ubx protein materials for enhanced purification of antibodies | |
WO2015093507A1 (ja) | プロテインgの細胞膜外ドメインの新規な改変型タンパク質 | |
JPWO2018180204A1 (ja) | 安定性改良型免疫グロブリン結合性ペプチド | |
JPWO2018180205A1 (ja) | 免疫グロブリン精製用アフィニティー分離マトリックス |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20111129 |
|
A601 | Written request for extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A601 Effective date: 20120227 |
|
A602 | Written permission of extension of time |
Free format text: JAPANESE INTERMEDIATE CODE: A602 Effective date: 20120301 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20120528 |
|
A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20130205 |
|
RD02 | Notification of acceptance of power of attorney |
Free format text: JAPANESE INTERMEDIATE CODE: A7422 Effective date: 20130213 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20130604 |
|
A911 | Transfer to examiner for re-examination before appeal (zenchi) |
Free format text: JAPANESE INTERMEDIATE CODE: A911 Effective date: 20130723 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20131008 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20140107 |
|
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: 20140128 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20140214 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 5480558 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
S531 | Written request for registration of change of domicile |
Free format text: JAPANESE INTERMEDIATE CODE: R313531 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |