WO2013168965A2 - 포도상구균 감염 예방용 백신 조성물 - Google Patents
포도상구균 감염 예방용 백신 조성물 Download PDFInfo
- Publication number
- WO2013168965A2 WO2013168965A2 PCT/KR2013/003957 KR2013003957W WO2013168965A2 WO 2013168965 A2 WO2013168965 A2 WO 2013168965A2 KR 2013003957 W KR2013003957 W KR 2013003957W WO 2013168965 A2 WO2013168965 A2 WO 2013168965A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- wta
- phosphate
- ribi
- glcnac
- modified
- Prior art date
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 31
- 229960005486 vaccine Drugs 0.000 title claims abstract description 22
- 206010041925 Staphylococcal infections Diseases 0.000 title claims abstract description 17
- 208000015339 staphylococcus aureus infection Diseases 0.000 title abstract 3
- OVRNDRQMDRJTHS-UHFFFAOYSA-N N-acelyl-D-glucosamine Natural products CC(=O)NC1C(O)OC(CO)C(O)C1O OVRNDRQMDRJTHS-UHFFFAOYSA-N 0.000 claims abstract description 25
- MBLBDJOUHNCFQT-LXGUWJNJSA-N N-acetylglucosamine Natural products CC(=O)N[C@@H](C=O)[C@@H](O)[C@H](O)[C@H](O)CO MBLBDJOUHNCFQT-LXGUWJNJSA-N 0.000 claims abstract description 25
- 229950006780 n-acetylglucosamine Drugs 0.000 claims abstract description 18
- OVRNDRQMDRJTHS-FMDGEEDCSA-N N-acetyl-beta-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-FMDGEEDCSA-N 0.000 claims abstract description 17
- 239000002253 acid Substances 0.000 claims abstract description 11
- 239000004480 active ingredient Substances 0.000 claims abstract description 6
- OVRNDRQMDRJTHS-RTRLPJTCSA-N N-acetyl-D-glucosamine Chemical compound CC(=O)N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-RTRLPJTCSA-N 0.000 claims abstract 11
- 229910019142 PO4 Inorganic materials 0.000 claims description 93
- 239000010452 phosphate Substances 0.000 claims description 93
- 241000191967 Staphylococcus aureus Species 0.000 claims description 32
- 238000000034 method Methods 0.000 claims description 30
- 241000191940 Staphylococcus Species 0.000 claims description 29
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 25
- 230000035772 mutation Effects 0.000 claims description 20
- 208000015181 infectious disease Diseases 0.000 claims description 17
- 241000295644 Staphylococcaceae Species 0.000 claims description 14
- 108090000623 proteins and genes Proteins 0.000 claims description 12
- 102000004190 Enzymes Human genes 0.000 claims description 10
- 108090000790 Enzymes Proteins 0.000 claims description 10
- 206010062255 Soft tissue infection Diseases 0.000 claims description 8
- 239000003814 drug Substances 0.000 claims description 7
- 238000012986 modification Methods 0.000 claims description 7
- 230000004048 modification Effects 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 206010040872 skin infection Diseases 0.000 claims description 5
- 241000124008 Mammalia Species 0.000 claims description 4
- RJQXTJLFIWVMTO-TYNCELHUSA-N Methicillin Chemical compound COC1=CC=CC(OC)=C1C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 RJQXTJLFIWVMTO-TYNCELHUSA-N 0.000 claims description 4
- 206010035664 Pneumonia Diseases 0.000 claims description 4
- 206010057190 Respiratory tract infections Diseases 0.000 claims description 4
- 206010040047 Sepsis Diseases 0.000 claims description 4
- 230000000903 blocking effect Effects 0.000 claims description 4
- 238000000502 dialysis Methods 0.000 claims description 4
- 230000003053 immunization Effects 0.000 claims description 4
- 229960003085 meticillin Drugs 0.000 claims description 4
- 230000001717 pathogenic effect Effects 0.000 claims description 4
- 230000001154 acute effect Effects 0.000 claims description 3
- 230000002401 inhibitory effect Effects 0.000 claims description 3
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 claims description 3
- 230000002265 prevention Effects 0.000 claims description 3
- 238000001356 surgical procedure Methods 0.000 claims description 3
- 238000002649 immunization Methods 0.000 claims description 2
- 101710098119 Chaperonin GroEL 2 Proteins 0.000 claims 1
- 108090001060 Lipase Proteins 0.000 claims 1
- 102000004882 Lipase Human genes 0.000 claims 1
- 239000004367 Lipase Substances 0.000 claims 1
- 230000005764 inhibitory process Effects 0.000 claims 1
- 210000003734 kidney Anatomy 0.000 claims 1
- 235000019421 lipase Nutrition 0.000 claims 1
- VJDOAZKNBQCAGE-LMVFSUKVSA-N D-ribitol 5-phosphate Chemical compound OC[C@H](O)[C@H](O)[C@H](O)COP(O)(O)=O VJDOAZKNBQCAGE-LMVFSUKVSA-N 0.000 abstract 2
- 230000008878 coupling Effects 0.000 abstract 1
- 238000010168 coupling process Methods 0.000 abstract 1
- 238000005859 coupling reaction Methods 0.000 abstract 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 22
- 230000027455 binding Effects 0.000 description 22
- 210000002966 serum Anatomy 0.000 description 21
- 235000021317 phosphate Nutrition 0.000 description 19
- 230000008021 deposition Effects 0.000 description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 12
- 206010057249 Phagocytosis Diseases 0.000 description 12
- 230000008782 phagocytosis Effects 0.000 description 12
- 239000011780 sodium chloride Substances 0.000 description 12
- 241000894006 Bacteria Species 0.000 description 11
- 230000001404 mediated effect Effects 0.000 description 11
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 10
- 230000002950 deficient Effects 0.000 description 10
- 210000003622 mature neutrocyte Anatomy 0.000 description 10
- 238000002360 preparation method Methods 0.000 description 10
- 239000006228 supernatant Substances 0.000 description 10
- 101150083828 tarS gene Proteins 0.000 description 10
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- 239000000243 solution Substances 0.000 description 8
- 238000005406 washing Methods 0.000 description 8
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 7
- OVRNDRQMDRJTHS-PVFLNQBWSA-N N-acetyl-alpha-D-glucosamine Chemical compound CC(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-PVFLNQBWSA-N 0.000 description 7
- 108700022034 Opsonin Proteins Proteins 0.000 description 7
- 239000002671 adjuvant Substances 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 7
- 229940098773 bovine serum albumin Drugs 0.000 description 7
- 239000000872 buffer Substances 0.000 description 7
- 230000024203 complement activation Effects 0.000 description 7
- 239000008188 pellet Substances 0.000 description 7
- 239000011269 tar Substances 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- QNAYBMKLOCPYGJ-UHFFFAOYSA-N D-alpha-Ala Natural products CC([NH3+])C([O-])=O QNAYBMKLOCPYGJ-UHFFFAOYSA-N 0.000 description 6
- MSFSPUZXLOGKHJ-UHFFFAOYSA-N Muraminsaeure Natural products OC(=O)C(C)OC1C(N)C(O)OC(CO)C1O MSFSPUZXLOGKHJ-UHFFFAOYSA-N 0.000 description 6
- 108010013639 Peptidoglycan Proteins 0.000 description 6
- 238000005119 centrifugation Methods 0.000 description 6
- 229940088598 enzyme Drugs 0.000 description 6
- 229940008228 intravenous immunoglobulins Drugs 0.000 description 6
- 239000013612 plasmid Substances 0.000 description 6
- 101150043888 tarM gene Proteins 0.000 description 6
- QNAYBMKLOCPYGJ-UWTATZPHSA-N D-alanine Chemical compound C[C@@H](N)C(O)=O QNAYBMKLOCPYGJ-UWTATZPHSA-N 0.000 description 5
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 5
- 101710088470 Immunoglobulin G-binding protein A Proteins 0.000 description 5
- 239000007983 Tris buffer Substances 0.000 description 5
- 239000011717 all-trans-retinol Substances 0.000 description 5
- 235000019169 all-trans-retinol Nutrition 0.000 description 5
- 230000000295 complement effect Effects 0.000 description 5
- 238000000684 flow cytometry Methods 0.000 description 5
- 238000009472 formulation Methods 0.000 description 5
- 239000008213 purified water Substances 0.000 description 5
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 5
- 239000011534 wash buffer Substances 0.000 description 5
- 229920000936 Agarose Polymers 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 4
- 101100368828 Staphylococcus aureus (strain COL) tarM gene Proteins 0.000 description 4
- 239000000427 antigen Substances 0.000 description 4
- 108091007433 antigens Proteins 0.000 description 4
- 102000036639 antigens Human genes 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 230000007812 deficiency Effects 0.000 description 4
- 230000001419 dependent effect Effects 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 101150096961 dltA gene Proteins 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 238000007911 parenteral administration Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 3
- OTMSDBZUPAUEDD-UHFFFAOYSA-N Ethane Chemical compound CC OTMSDBZUPAUEDD-UHFFFAOYSA-N 0.000 description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- DUKURNFHYQXCJG-UHFFFAOYSA-N Lewis A pentasaccharide Natural products OC1C(O)C(O)C(C)OC1OC1C(OC2C(C(O)C(O)C(CO)O2)O)C(NC(C)=O)C(OC2C(C(OC3C(OC(O)C(O)C3O)CO)OC(CO)C2O)O)OC1CO DUKURNFHYQXCJG-UHFFFAOYSA-N 0.000 description 3
- 108090001030 Lipoproteins Proteins 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- OVRNDRQMDRJTHS-ZTVVOAFPSA-N N-acetyl-D-mannosamine Chemical compound CC(=O)N[C@@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-ZTVVOAFPSA-N 0.000 description 3
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 3
- 102000004357 Transferases Human genes 0.000 description 3
- 108090000992 Transferases Proteins 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 210000002421 cell wall Anatomy 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003937 drug carrier Substances 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 3
- -1 hydroxypropyl Chemical group 0.000 description 3
- 229920001282 polysaccharide Polymers 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 229910052709 silver Inorganic materials 0.000 description 3
- 239000004332 silver Substances 0.000 description 3
- 239000001509 sodium citrate Substances 0.000 description 3
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 3
- 239000002904 solvent Substances 0.000 description 3
- 101150020588 tagO gene Proteins 0.000 description 3
- 238000002255 vaccination Methods 0.000 description 3
- ZUQUTHURQVDNKF-WZPXOXCRSA-N 1-[(3S,4R,5S,6R)-3-amino-2,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]ethanone Chemical compound C(C)(=O)C1(O)[C@@H](N)[C@@H](O)[C@H](O)[C@H](O1)CO ZUQUTHURQVDNKF-WZPXOXCRSA-N 0.000 description 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 2
- 108010088751 Albumins Proteins 0.000 description 2
- 102000009027 Albumins Human genes 0.000 description 2
- 208000002109 Argyria Diseases 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 102000014914 Carrier Proteins Human genes 0.000 description 2
- 108010078791 Carrier Proteins Proteins 0.000 description 2
- 102000002322 Egg Proteins Human genes 0.000 description 2
- 108010000912 Egg Proteins Proteins 0.000 description 2
- 102100027723 Endogenous retrovirus group K member 6 Rec protein Human genes 0.000 description 2
- ULGZDMOVFRHVEP-RWJQBGPGSA-N Erythromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@@](C)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 ULGZDMOVFRHVEP-RWJQBGPGSA-N 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- 102000051366 Glycosyltransferases Human genes 0.000 description 2
- 108700023372 Glycosyltransferases Proteins 0.000 description 2
- 101000580925 Homo sapiens Endogenous retrovirus group K member 6 Rec protein Proteins 0.000 description 2
- 102000004867 Hydro-Lyases Human genes 0.000 description 2
- 108090001042 Hydro-Lyases Proteins 0.000 description 2
- 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 2
- 102000004895 Lipoproteins Human genes 0.000 description 2
- 108700018351 Major Histocompatibility Complex Proteins 0.000 description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 2
- 102000016943 Muramidase Human genes 0.000 description 2
- 108010014251 Muramidase Proteins 0.000 description 2
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 2
- OVRNDRQMDRJTHS-OZRXBMAMSA-N N-acetyl-beta-D-mannosamine Chemical compound CC(=O)N[C@@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O OVRNDRQMDRJTHS-OZRXBMAMSA-N 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 101150066443 TARS1 gene Proteins 0.000 description 2
- 238000000516 activation analysis Methods 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 238000012790 confirmation Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 238000012217 deletion Methods 0.000 description 2
- 230000037430 deletion Effects 0.000 description 2
- 235000014103 egg white Nutrition 0.000 description 2
- 210000000969 egg white Anatomy 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 210000000222 eosinocyte Anatomy 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 108700014210 glycosyltransferase activity proteins Proteins 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 229910052816 inorganic phosphate Inorganic materials 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 150000002540 isothiocyanates Chemical class 0.000 description 2
- 108010045069 keyhole-limpet hemocyanin Proteins 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 239000004325 lysozyme Substances 0.000 description 2
- 229960000274 lysozyme Drugs 0.000 description 2
- 235000010335 lysozyme Nutrition 0.000 description 2
- 101150020801 oatA gene Proteins 0.000 description 2
- 229950008882 polysorbate Drugs 0.000 description 2
- 229920000136 polysorbate Polymers 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 101150065015 spa gene Proteins 0.000 description 2
- 230000009870 specific binding Effects 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 230000020382 suppression by virus of host antigen processing and presentation of peptide antigen via MHC class I Effects 0.000 description 2
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 1
- CYDQOEWLBCCFJZ-UHFFFAOYSA-N 4-(4-fluorophenyl)oxane-4-carboxylic acid Chemical compound C=1C=C(F)C=CC=1C1(C(=O)O)CCOCC1 CYDQOEWLBCCFJZ-UHFFFAOYSA-N 0.000 description 1
- YRNWIFYIFSBPAU-UHFFFAOYSA-N 4-[4-(dimethylamino)phenyl]-n,n-dimethylaniline Chemical compound C1=CC(N(C)C)=CC=C1C1=CC=C(N(C)C)C=C1 YRNWIFYIFSBPAU-UHFFFAOYSA-N 0.000 description 1
- 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 1
- 244000215068 Acacia senegal Species 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-M Bicarbonate Chemical compound OC([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-M 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 102000009016 Cholera Toxin Human genes 0.000 description 1
- 108010049048 Cholera Toxin Proteins 0.000 description 1
- 108010028780 Complement C3 Proteins 0.000 description 1
- 102000016918 Complement C3 Human genes 0.000 description 1
- 108010028778 Complement C4 Proteins 0.000 description 1
- 150000008541 D-alanines Chemical class 0.000 description 1
- 101710146739 Enterotoxin Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- QUSNBJAOOMFDIB-UHFFFAOYSA-N Ethylamine Chemical compound CCN QUSNBJAOOMFDIB-UHFFFAOYSA-N 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101100450591 Human adenovirus B serotype 3 PVIII gene Proteins 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 102000009490 IgG Receptors Human genes 0.000 description 1
- 108010073807 IgG Receptors Proteins 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- 108090000988 Lysostaphin Proteins 0.000 description 1
- 101100348848 Mus musculus Notch4 gene Proteins 0.000 description 1
- FXHOOIRPVKKKFG-UHFFFAOYSA-N N,N-Dimethylacetamide Chemical compound CN(C)C(C)=O FXHOOIRPVKKKFG-UHFFFAOYSA-N 0.000 description 1
- MBBZMMPHUWSWHV-BDVNFPICSA-N N-methylglucamine Chemical compound CNC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO MBBZMMPHUWSWHV-BDVNFPICSA-N 0.000 description 1
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 1
- 229920002685 Polyoxyl 35CastorOil Polymers 0.000 description 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 1
- 238000002105 Southern blotting Methods 0.000 description 1
- 241000823609 Staphylococcus aureus subsp. aureus RN4220 Species 0.000 description 1
- 230000006052 T cell proliferation Effects 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- LFTYTUAZOPRMMI-CFRASDGPSA-N UDP-N-acetyl-alpha-D-glucosamine Chemical compound O1[C@H](CO)[C@@H](O)[C@H](O)[C@@H](NC(=O)C)[C@H]1OP(O)(=O)OP(O)(=O)OC[C@@H]1[C@@H](O)[C@@H](O)[C@H](N2C(NC(=O)C=C2)=O)O1 LFTYTUAZOPRMMI-CFRASDGPSA-N 0.000 description 1
- LFTYTUAZOPRMMI-UHFFFAOYSA-N UNPD164450 Natural products O1C(CO)C(O)C(O)C(NC(=O)C)C1OP(O)(=O)OP(O)(=O)OCC1C(O)C(O)C(N2C(NC(=O)C=C2)=O)O1 LFTYTUAZOPRMMI-UHFFFAOYSA-N 0.000 description 1
- 206010000269 abscess Diseases 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 108020002494 acetyltransferase Proteins 0.000 description 1
- 102000005421 acetyltransferase Human genes 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 125000003295 alanine group Chemical group N[C@@H](C)C(=O)* 0.000 description 1
- PPQRONHOSHZGFQ-LMVFSUKVSA-N aldehydo-D-ribose 5-phosphate Chemical compound OP(=O)(O)OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PPQRONHOSHZGFQ-LMVFSUKVSA-N 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000003855 balanced salt solution Substances 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 229920002988 biodegradable polymer Polymers 0.000 description 1
- 239000004621 biodegradable polymer Substances 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
- 239000004359 castor oil Substances 0.000 description 1
- 235000019438 castor oil Nutrition 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 239000003221 ear drop Substances 0.000 description 1
- 229940047652 ear drops Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000147 enterotoxin Substances 0.000 description 1
- 231100000655 enterotoxin Toxicity 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 229960003276 erythromycin Drugs 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 239000003889 eye drop Substances 0.000 description 1
- 229940012356 eye drops Drugs 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 150000004676 glycans Polymers 0.000 description 1
- RBNPOMFGQQGHHO-UHFFFAOYSA-N glyceric acid Chemical class OCC(O)C(O)=O RBNPOMFGQQGHHO-UHFFFAOYSA-N 0.000 description 1
- 125000005639 glycero group Chemical group 0.000 description 1
- AWUCVROLDVIAJX-UHFFFAOYSA-N glycerol 1-phosphate Chemical class OCC(O)COP(O)(O)=O AWUCVROLDVIAJX-UHFFFAOYSA-N 0.000 description 1
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 1
- 229920000550 glycopolymer Polymers 0.000 description 1
- 230000001279 glycosylating effect Effects 0.000 description 1
- 244000000059 gram-positive pathogen Species 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000005745 host immune response Effects 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000000984 immunochemical effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 239000007951 isotonicity adjuster Substances 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000012669 liquid formulation Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- DKXULEFCEORBJK-UHFFFAOYSA-N magnesium;octadecanoic acid Chemical compound [Mg].CCCCCCCCCCCCCCCCCC(O)=O DKXULEFCEORBJK-UHFFFAOYSA-N 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 229960003194 meglumine Drugs 0.000 description 1
- WVJKHCGMRZGIJH-UHFFFAOYSA-N methanetriamine Chemical compound NC(N)N WVJKHCGMRZGIJH-UHFFFAOYSA-N 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 239000007923 nasal drop Substances 0.000 description 1
- 229940100662 nasal drops Drugs 0.000 description 1
- 210000000440 neutrophil Anatomy 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- QUANRIQJNFHVEU-UHFFFAOYSA-N oxirane;propane-1,2,3-triol Chemical compound C1CO1.OCC(O)CO QUANRIQJNFHVEU-UHFFFAOYSA-N 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 1
- 239000008024 pharmaceutical diluent Substances 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- QVLTXCYWHPZMCA-UHFFFAOYSA-N po4-po4 Chemical group OP(O)(O)=O.OP(O)(O)=O QVLTXCYWHPZMCA-UHFFFAOYSA-N 0.000 description 1
- 239000008389 polyethoxylated castor oil Substances 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 150000004804 polysaccharides Polymers 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000001540 sodium lactate Substances 0.000 description 1
- 229940005581 sodium lactate Drugs 0.000 description 1
- 235000011088 sodium lactate Nutrition 0.000 description 1
- NGXXZCXLGGIIKN-UHFFFAOYSA-M sodium;ethane-1,2-diamine;hydroxide Chemical compound [OH-].[Na+].NCCN NGXXZCXLGGIIKN-UHFFFAOYSA-M 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 230000000699 topical effect Effects 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 230000014621 translational initiation Effects 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/02—Bacterial antigens
- A61K39/085—Staphylococcus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
- A61K2039/575—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 humoral response
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
Definitions
- Vaccine composition for preventing staphylococcal infection
- the present invention relates to a vaccine composition for preventing staphylococcal infection, and more particularly, to ribibisphosphate or a repeating unit thereof, wherein N-acetylglucosamine (GlcNAc) is modified to be ⁇ -position only, or It relates to a vaccine composition for preventing staphylococcal infection, which contains the above-mentioned repeat teychoic acid (WTA) as an active ingredient.
- WTA repeat teychoic acid
- Staphylococcus is a Gram-positive pathogen with a single cell membrane surrounded by glycopolymers including wall teichoic acid (WTA), peptidoglycan, lipoteicoic acid and capsular polysacchar ide (FIG. 1). Reference).
- Staphylococcus aureus WTA is covalently bound to peptidoglycan and contains N-acetylmannosamine (ManNAc)-(
- ManNAc N-acetylmannosamine
- GlcNAc acetylglucosamine
- the hydroxyl group on the ribi-phosphate repeat unit in the above structure is cationic ealanine.
- Ester and GlcNAc Xia G et al., Int. J. Med. Microbiol., 300: 148 ′ 154, 2010; see FIG. 2).
- Staphylococcus WTA is not essential for bacterial survival, but is known to be involved in nasal epithelial cell adhesion (Weidenmaier C, et al., Nature medicine, 10: 243-245, 2004). Staphylococcus aureus WTA is also known to regulate abscess formation in mouse skin infection models by inducing CD4 + T-cell proliferation in a major histocompatibility complex (MHC) II-dependent manner (Weidenmaier C et al. , PloS one 5: el3227, 2010). To analyze the chemical and antigenic structures of the cell walls of Staphylococcus Copenhagen strains in the early 1960s, several groups performed immunochemical studies using rabbit serum.
- MHC major histocompatibility complex
- the TarM and TarS enzymes have been found to be capable of glycosylating ⁇ -GlcNAc and ⁇ -GlcNAc residues to libri-phosphate in WTA, respectively, in a UDP-GlcNAc-dependent manner. While we studied the WTA of Staphylococcus aureus, the ⁇ -GlcNAc residue of Staphylococcus aureus induces classical complement-dependent opsonophagocytosis and recognizes ⁇ -GlcNAc residue of WTA as epitope in serum. The present invention was completed by revealing that the antibody to be produced is mainly produced. SUMMARY OF THE INVENTION Accordingly, it is an object of the present invention to provide a vaccine composition for preventing staphylococcal infection.
- Another object of the present invention is to provide a method for producing anti-WTA antibodies of Staphylococcus aureus.
- a native WTA comprising a ribi-phosphate repeat unit modified with N-acetylglucosamine (GlcNAc)
- GlcNAc is not modified with the a-configuration to the ribi-phosphate, A ribi-phosphate modified only at the position ( ⁇ -configuration)
- GlcNAc is a repeating unit of ribi-phosphate modified at the ⁇ -position, not modified at the ⁇ -position to the ribi-phosphate, or
- iii) GlcNAc is not modified at the ⁇ ′ position to the ribi-phosphate, but modified at the ⁇ ′ position.
- a vaccine composition for preventing staphylococcal infection comprising WTA containing a repeating unit of ribi-phosphate as an active ingredient.
- the present invention (1) mutant Staphylococcus aureus containing the natural wall teichoic acid (WTA) modified with N-acetyl glucosamine (GlcNAc) is N ⁇ acetylglucosamine (GlcNAc) ⁇
- WTA natural wall teichoic acid
- GlcNAc N-acetyl glucosamine
- GlcNAc N acetylglucosamine
- FIG. 2 is a schematic diagram showing the structure of Staphylococcus aureus teicosane (WTA).
- FIG 3 is a schematic diagram showing the WTA of Staphylococcus aureus, the residues modified on the WTA and related genes.
- GlcNAc is an abbreviation of N-acetylglucosamine
- ManNAc is an abbreviation of N ⁇ acetylmannosamine
- Ala is an abbreviation of D-alanine
- Pi is an abbreviation of phosphate.
- Figure 4 shows the binding between mutant strains and anti-WTA antibodies and C3 and C4 deposition in Staphylococcus strains and mutations (liar 5, ⁇ tarS, A tarM, Adit A, and zl ⁇ ⁇ ⁇ (9) according to the present invention This is confirmed by flow cytometry.
- Figure 5 shows the binding capacity of the anti-WTA antibody and the C3 and C4 activation analysis according to the concentration of the anti—WTA antibody.
- Figure 6 confirms the induction of complement-mediated townson phagocytosis of the parent strain and the mutant strain according to the present invention, it is the result of measuring the number of bacteria in 100 human polymorphonuclear neutrophil (PMN).
- PMN human polymorphonuclear neutrophil
- WTA Warteichoic acid
- ManNAc N-acetylmannosamine
- GkNAc N-acetylglucosamine
- the ribitose phosphate is modified with N—acetylglucosamine (GlcNAc) and D-alanine, which GlyCac has the ⁇ -position and / Or ⁇ -configuration.
- GlcNAc N—acetylglucosamine
- D-alanine which GlyCac has the ⁇ -position and / Or ⁇ -configuration.
- ⁇ -GlcNAc deficiency or "a-GlcNAcylation deficiency” refers to a condition in which GlcNAc does not bind to the a-position of ribidyl phosphate in WTA
- ⁇ -GlcNAc deficiency or “crab deficiency” refers to a condition in which GlcNAc is unable to bind libby at the ⁇ -position of phosphate-phosphate in WTA.
- anti-WTA antibody refers to an antibody that specifically binds WTA, which induces complement-mediated opsonophagocytosis to prevent infection from Staphylococcus aureus. To protect the host.
- the present invention starts from the identification of WTA binding motifs (epitopes) recognized by anti-WTA antibodies.
- the present invention relates to mutations lacking the tarM and / or tarS genes associated with modifications of a -GlcNAc and ⁇ -GlcNAc on WTA of Staphylococcus aureus, and mutations AtarS and AtarMS lacking both residues of a -GlcNAc and ⁇ -GlcNAc).
- Various experiments used suggest that GlcNAc residues that bind livi in the WTA—the phosphate to the ⁇ -position are major epitopes for anti-WTA antibodies that induce complement-mediated opsonin phagocytosis.
- the present invention binds the ⁇ -GlcNAcylated WTA isolated from staphylococcus parent strains, / arM mutant strains and ⁇ mutant strains, but not the tadiS double mutant strain, liarS mutant strains, and / tag0 mutations.
- ⁇ -GlcNAc isolated from strain does not bind to unmodified WTA (see Example 2).
- the present invention is characterized by the presence of C3 and C4 deposits on the parent strain, mutant strain, and ⁇ ( ⁇ mutant strains) of Staphylococcus aureus having ⁇ -GlcNAcylated WTA, while having ⁇ -GlcNAc unconverted WTA.
- the present invention provides the fact that anti-WTA antibodies bind to GlcNAc residues modified with native WTA-increasing ⁇ -positions that are modified in the WTA to ⁇ - and / or ⁇ -positions to cause host immune response.
- the present invention provides a natural WTA containing a ribo-phosphate repeat unit modified with N-acetylglucosamine (GlcNAc), wherein (i) GlcNAc is modified with the ⁇ -position ( ⁇ ⁇ configuration) to the ribi-phosphate. Ribi-phosphate modified only at the ⁇ -position; (ii) GlcNAc is modified to the ribi-phosphate at the ⁇ -position but not at the ⁇ -position.
- Repeating unit, or (Hi) Staphylococcus aureus containing, as an active ingredient, WTA containing a repeating unit of a ribi-phosphate modified at the ⁇ -position but not modified at the ribi-phosphate in the ⁇ -position.
- a vaccine composition for preventing infection.
- GlcNAc which is used as an active ingredient in the vaccine composition according to the present invention, is not modified by the ⁇ -position (a-configuration) to the ribi ⁇ phosphate, but is modified by the ⁇ -position ( ⁇ ⁇ configuration) — A phosphate, (ii) a repeating unit of ribi-phosphate that GlcNAc is not modified in the ribi-phosphate at the ⁇ -position, but modified only at the ⁇ -position, or (iii) GlcNAc is ⁇ - in the ribi-phosphate WTAs that contain repeat units of Revital-Phosphate modified only in the ⁇ -position, but not in position, modify GlcNAc to the ⁇ -position of the Livi-phosphate in WTA of Staphylococcus ((1-( ⁇ 1 1010) by inhibiting the function of TarM enzymes to block the modification of GlcNAc in the ⁇ -position to the ribi-phosphate in WTA
- a-position
- the vaccine composition according to the present invention is an infection of Staphylococcus, specifically infection due to the use of a catheter during renal dialysis or infection during surgery, skin or soft tissue infection (SSTI), pneumonia, sepsis or acute respiratory tract infect can be used for the prevention of ions).
- SSTI skin or soft tissue infection
- the staphylococci may be, but are not limited to, methicillin-resistant staphylococcus aureus (MRSA) or pathogenic staphylococci.
- the vaccine composition according to the present invention may further include a pharmaceutically acceptable carrier, diluent and / or adjuvant in addition to the WTA.
- the carrier used in the vaccine composition according to the present invention is selected based on the method and route of administration and the standard drug composition, for example carrier protein (ie bovine serum albumin (BSA), egg white albumin (OVA), human serum). Albumin (HSA) and keyhole limpet hemocyanin (KLH)), solubilizers (ie, ethane, polysorbate and Cremophor EL TM), isotonic agents, preservatives, antioxidant excipients (ie, lactose, starch, crystalline sallolo) agarose, agarose and the manni, a maltose, calcium hydrogen "kalseum, gyeongmu can silicate and carbonate kalseum), the binder (i.e., starch, polyvinyl pyrrolidone, hydroxypropyl as hydroxypropyl selreul agarose, with ethyl selreul agarose, carboxymethyl selreul Gum arab
- the vaccine composition according to the present invention may be combined with a solution (1) known as a carrier protein (Calbiotec, 50 mg of glycerol dissolved in 125 mg per 1 ml of solution) to enhance antigenicity.
- a carrier protein Calbiotec, 50 mg of glycerol dissolved in 125 mg per 1 ml of solution
- Diluents used in the vaccine composition according to the present invention may be selected based on the method and route of administration and the actual standard drug composition.
- diluents include water, saline, phosphate complete saline and bicarbonate solutions.
- Adjuvants used in the vaccine composition according to the invention are selected based on the method and route of administration and the actual standard drug composition. Examples of adjuvant include cholera toxin, bipyretic enterotoxin (LT) of E. coli, liposomes and immune stimulatory complex (ISC0M).
- the route of administration may vary depending on the age, body weight, sex and general health of the subject with the risk of staphylococcal infections, but administration may be oral and parenteral (eg, intravenous, arterial and topical). It can be administered by any of the routes. Especially, parenteral administration is preferable.
- Formulations for oral and parenteral administration and methods for their preparation are known to those skilled in the art.
- Formulations for oral administration and parenteral administration may be prepared by conventional procedures, for example, in combination with the aforementioned pharmaceutically acceptable carriers.
- formulations for oral administration include solid or liquid formulations such as solvents, tablets, granules, powders or capsules.
- formulations for parenteral administration include solvents, suspensions, ointments, creams, suppositories, eye drops, nasal drops and ear drops.
- biodegradable polymers eg, poly-D, L-lactide-co-glycoside or polyglycoside
- buck base eg, US patents. 5, 417,986, 4,675,381 and 4, 450,150
- flavors and colorings may be added.
- Suitable pharmaceutical carriers, diluents and pharmaceutically necessary substances for their use are described in Remington's Pharmaceutical Sciences.
- the dosage of the vaccine composition according to the present invention is determined based on the type of adjuvant, the method and frequency of administration, and the desired effect and may generally be WTA l // g to lOOmg for single adult administration.
- the dosage may generally be from WTA 1 / ig to lmg in one adult dose.
- the administration can be administered several times if necessary. For example, initial vaccination followed by three booster vaccinations can be performed at weekly intervals. Alternatively, adjuvant injection and second adjuvant vaccination may be performed at week 8-12 and week 16-20, respectively, from the first immunization using the same agent.
- the present invention (1) mutated Staphylococcus aureus containing natural wall teichoic acid (WTA) modified with N-acetylglucosamine (GlcNAc), whereby N-acetylglucosamine (GlcNAc) is in the ⁇ -position ( ⁇ -configuration).
- Step (1) mutates the native Staphylococcus aureus containing the native wall teichoic acid (WTA) modified with ⁇ -acetylglucosamine (GlcNAc), whereby N-acetylglucosamine (GlcNAc) is in the ⁇ -position ( ⁇ -configuration )
- WTA native wall teichoic acid
- GlcNAc ⁇ -acetylglucosamine
- the procedure of step (1) can be accomplished by blocking the function of Staphylococcus TarM enzymes to modify the ribi in ⁇ —to modify the GlcNAc in ⁇ -position to phosphate, as described above.
- the tarM gene encoding the TarM enzyme in a M0107 strain lacking IgG binding protein A (see Oku Y et al., Journal of bacteriology, 191: 141 ⁇ 151, 2009) Deletion results in a mutant strain lacking ⁇ -GkNAc.
- step (2) is performed by (i) GlcNAc from the mutant Staphylococcus aureus, which is not modified by the a-position to the ribo-phosphate, but by the ⁇ -position only. Modified ribi—phosphate (ii) GlcNAc modifies the libi—in the phosphate, not modified at the ⁇ -position, but modified at the ⁇ -position Repeating units of ribi-phosphate, or (iii) GlcNAc isolating and purifying WTA comprising repeating units of ribi-phosphate modified only at the ⁇ -position but not modified at the ⁇ -position to the ribi-phosphate. Step.
- the process can be carried out according to methods known in the art, for example according to the methods of Shitatsuchi A et al., I'unology, 129: 268-277, 2010 or by using the same. It is also possible to obtain WTA from which it is possible to further separate and purify only the repeat units of the ribi-phosphate, or also to further separate and purify the ribi-phosphate only from the repeat units of the ribi-phosphate. .
- step (3) further comprises the steps of: anti-WTA using the separated and purified ribi-phosphate repeating units of the ribi-phosphate or WTA comprising the repeating unit of the ribi-phosphate.
- the process of producing antibodies can be carried out using a variety of methods known in the art for producing antibodies from antigens, for example the isolated and purified ribi-phosphate, repeating units of the ribi-phosphate, or the rev -Can be carried out by immunizing a mammal with a WTA comprising a repeat unit of phosphate.
- the anti-WTA antibodies prepared above may be infected with Staphylococcus, for example, infection due to the use of a catheter during renal dialysis or infection during surgery, skin or soft tissue infection (SSTI), pneumonia, sepsis or acute respiratory infection. It can be used to prevent tract infection.
- Staphylococcus for example, infection due to the use of a catheter during renal dialysis or infection during surgery, skin or soft tissue infection (SSTI), pneumonia, sepsis or acute respiratory infection. It can be used to prevent tract infection.
- the aforementioned staphylococci can be, but are not limited to, methicillin-resistant staphylococci (MRSA) or pathogenic staphylococci.
- the present invention also provides a WTA or step (3) comprising a ribi-phosphate separated and purified in step (2), the ribi-repeating unit of phosphate, or a repeating unit of the ribose-phosphate.
- a method of preventing staphylococcal infection comprising administering an anti-WTA antibody to a mammal.
- the present invention provides a use for the preparation of a medicament for preventing staphylococcal infection of the WTA or the anti-WTA antibody produced in step (3) comprising the repeat unit of the ribo-phosphate, or the repeat unit of the ribo-phosphate.
- the present invention will be described in detail with reference to Examples, which are merely illustrative of the present invention, and thus the scope of the present invention is not limited thereto.
- Staphylococcus aureus acidic is a livi substituted with two glycerates followed by phosphate followed by ⁇ - or ⁇ -GlcNAc and D-alanine. It is bound to peptidoglycan, including acetylmannosamine (ManNAc) and acetylglucosamine (GlcNAc) disaccharide linker (see FIG. 3).
- staphylococci mutants lacking GlcNAc or D-alanine modified with ⁇ - or ⁇ -positions of ribi-phosphate in WTA were prepared.
- the RN4220 strain Prior to identifying epitopes for anti-WTA antibodies, the RN4220 strain (NovickRP ei a /., Embo. J., 12: 3967-, to prevent the anti-WTA antibodies from binding to IgG binding protein A in Staphylococcus aureus). 3975. 1993) produced a mutant strain (M0107) deficient in IgG-binding protein A by deleting the spa gene, a gene for IgG-binding protein A (Oku Y, et al., Journal of bacteriology, 191: 141). -151, 2009).
- the spa gene of the RN4220 strain was replaced with a Phleo resistance gene by double crossover recombinat ion using the following primers:
- spa-Pi GGGTCTAGAAAAAAGTCAAGCCTGAAGTCG (SEQ ID NO: 1); spa-P2: TATTGGATCC ⁇ GTGGGGCriTGAATGTG (SEQ ID NO: 2);
- spa-P3 CCCGGGTACCTGCAGCGTTATTAGCTGGAC (SEQ ID NO: 3);
- P4 GGGGMTTCTMTTGGTGCMCTGGGACA (SEQ ID NO: 4);
- Phleo-Pl GGATCCAATAGACCAGTTGCA (SEQ ID NO: 5);
- Phleo-P3 GGTACCCGGGCGATTGCTGAA (SEQ ID NO: 6).
- the mutant strain (T803) lacking ⁇ -GlcNAc was prepared by deleting the tarS gene from the M0107 strain prepared in Example ⁇ 1-1>.
- pSF151 3 ⁇ 4 lasmid (km r ) having the central open reading frame region of the tarS gene at the BamHI and EcoRI positions of the tarS gene encoding ⁇ -GlcNAc transferase (Tao L et al. , Gene, 120: 105-110, 1992) were inserted to destroy the i r5 gene.
- the disrupted tarS gene was amplified by PCR using the following primers and confirmed by electrophoresis:
- the mutant strain (T790) deficient in a-GkNAc was prepared by deleting the tarM gene from the M0107 strain prepared in Example ⁇ 1 ⁇ 1>.
- a pMut inT3 plasmid having a central open reading frame region of the tarM gene at the Hind ⁇ and Ban gene positions of the iar gene encoding a-GlcNAc transferase (Vagner). V et al., Microbiology (Reading, England) 144 (Pt 11): 3097-3104, 1998) were inserted to destroy the tarM gene.
- the disrupted tarM gene was amplified by PCR using the following primers and confirmed by electrophoresis.
- tad l-BaiM CACGGATCCTAAATGCACCCGTATCATCGAA (SEQ ID NO: 10) ⁇ 1-4>
- T807 Deficient in ⁇ -GlcNAc and ⁇ —GlcNAc
- the tarM and tarS genes were deleted from the M0107 strain prepared in Example ⁇ 1-1>, and thus, in WTA.
- a mutant strain ( ⁇ 807) was prepared in which ribi-phosphate lacked both GlcNAc modifications to the ⁇ - and ⁇ - positions.
- the mutations were prepared using the same method as in Examples ⁇ 1-2> and ⁇ 1-3>.
- the mutant strain (T861) deficient in D-alanine was prepared by deleting the dltA gene from the M0107 strain prepared in Example ⁇ 1-1>.
- the internal region of the ntA gene (positions 46 to 694; position 1 is a translation initiation site) was amplified by PCR and inserted into a pMutinT3 vector to prepare plasmid ⁇ 0793.
- the plasmid pTO793 was transformed into a M0107 strain.
- the erythromycin resistance strain was then selected to obtain a mutant strain (T861) in which the dltA gene was destroyed, the destruction of the dltA gene and insertion of the plasmid into the desired chromosomal locus by Southern blot.
- the mutant strain (T861) lacking WTA was prepared by deleting the tagO gene from the M0107 strain prepared in Example ⁇ 1-1>.
- plasmid pT0702 was prepared by amplifying the inner region of the tagO gene (positions 36 to 645) by PCR and inserting it into the pMutinT3 vector.
- the plasmid pT0793 was transformed into a M0107 strain to obtain a mutant strain (T861) in which the tagO gene was destroyed. Information on the aforementioned mutant strains is shown in Table 1 below.
- an anti-WTA antibody was prepared from WTA, and the binding between the antibody prepared above and the mutant strain prepared in Example 1 was analyzed.
- T363 strain (RN4220 Mgf.:phleo) and T002 Strain (RN4220 oatA enn) was obtained and T384 strain was prepared from phage 80 alpha transduction from them. Since the Igt mutation is removed by bacterial lipoproteins, it is possible to isolate WTA without lipoprotein contamination from this strain. The oatA mutation also makes staphylococcal peptidoglycan susceptible to lysozyme Oysozyme.
- the supernatant was removed by centrifugation at 6,000 rpm for 15 minutes.
- the pellet was suspended in 10 ml of sodium citrate (20 mM pH 4.5) and then centrifuged at 15,000 rpm for 5 minutes to remove the supernatant.
- the pellet was again suspended in 20 ml of sodium citrate (20113 ⁇ 4, 4.5) supplemented with lMNaCl, mixed with glass beads (12 g per 1 L) and then disrupted cells using a beads blender.
- the disrupted cells were transferred to a 50 ml tube, centrifuged at 3,000 rpm for 10 minutes to collect only the supernatant, followed by centrifugation at 15,000 rpm for 10 minutes to remove the supernatant and pellets.
- the pellet was suspended in 20 ml of 20 niM sodium citrate (pH 4.5) containing 0.5% SDS, boiled at 6 (C for 30 minutes, and centrifuged at 15,000 rpm for 10 minutes to remove the supernatant. After complete removal of SDS, washing was performed with 20 mM Tris (pH 7.0) to neutralize pH, and then the pellet was suspended in 5 ml of 20 mM Tris (pH 7.0), followed by 50 ⁇ l of 1M CaCl 2 and 50 ⁇ l of trypsin (20 mg / ml) was added and incubated for 12 hours at 37 ° C.
- the culture was then centrifuged at 15,000 rpm for 10 minutes to remove the supernatant, and the pellet was poured into 20 nil of distilled water for injection. Suspension and washing The pellets, repeated 5-6 times, were suspended in 1 ml of distilled water for injection and then lyophilized.
- the lyophilisate containing WTA was suspended in 1 ml of Tris (pH 7.0) per 50 mg and treated with lysostaphin (Sigma Aldrich; O.lmg per lOnig of lyophilisate) for 12 hours at 37 ° C.
- lysostaphin Sigma Aldrich; O.lmg per lOnig of lyophilisate
- O.lmg treatment per 10 mg of lyophilisate was added to react for 12 hours.
- the reaction mixture was heated at 95 ° C for 10 minutes to inactivate the enzyme, and then the supernatant was collected by centrifugation at 15,000 rpm.
- the harvested supernatant was filtered at 0.45 ⁇ .
- ion exchange chromatography was performed under the following conditions to purify only WTA to which the peptidoglycan monomer was attached. ⁇ Column: Hitrap Q FF TM 5ml
- Fractions obtained from the chromatography were mixed with cold acetone in a ratio of 1: 3 and precipitated, followed by centrifugation at 15,000 rpn] for 20 minutes to remove the supernatant. Subsequently, the precipitate was sprayed with acetone 40-50 ⁇ l and hardened, and then collected in a 1.5 ml tube and dried for 1 hour to remove acetone. The dried WTA was dissolved in 500 ⁇ l of distilled water for injection and then lyophilized.
- Anti-WTA antibodies were isolated and purified from commercial human intravenous immunoglobulins (IVIG, Green Cross, Korea, 400 mg total) using WTA obtained in Example ⁇ 2-1>.
- WTA was dropped onto a nitrocell membrane (10 ⁇ 90 ⁇ s, Whatman, pore 0.45 ⁇ ) and then baked at 100 ° C. for 1 hour.
- the membrane was washed with complete agent A (20 mM Tris-HCl, pH 7.4, 150 niM NaCl) and complete D (20 mM Tris-HCl, pH 7.4, 150 mM NaCl and 1% BSA) at 4 ° C for 2 hours. Blocked.
- the nitrocell membranes were incubated with 50 nig of IVIG dissolved in 40 ml of Buffer E (10 mM Tr is, pH 7.4, 140 niM NaCl) for 2 hours at 4 ° C.
- the antibody bound WTA was eluted with 1 ml 0.1 M glycine (pH 2.8) and the pH was neutralized to 7.5 directly with 1 M K0H. Thereafter, the glycine present in the eluate was removed by centrifugation three times with Vivaspin 20 (Sartorius) using the complete agent A. Thereafter, in order to remove the anti-peptidoglycan antibody, the obtained IgG fraction was added to formaldehyde pre-fixed at 4 ° C. with the T258 strain lspa and AtagO double mutant strain of Examples ⁇ 1-6>). Incubate for 2 hours to bind the strain and anti-peptidoglycan antibody. After centrifugation of the culture, the supernatant was concentrated by vivaspine 20 to obtain an anti-WTA antibody.
- the cultured strains were washed with wash buffer (10 mM Tris-HCKpH 7.4), 140 mM NaCl, 10 mM CaCl 2 and 0.05% Twenty 20). Then, the washed strain was sonicated for 15 seconds to disperse the collected strains, and then flow cytometry (Accuri C6, Beckman Coulter) was performed.
- Gray areas in Figures a to f are the results of culturing Staphylococcus mutant strains killed with ethane without anti-WTA IgG, and black lines incubating Staphylococcus mutant bacteria killed with ethanol with anti-WTA antibodies. The result is.
- purified anti-WTA-IgG is the parent strain M0107
- complement activation pathway binding of the antigen to the antibody results in complement activation, whereby deposition of complements C3 and C4 occurs. Therefore, by examining the presence of C3 and C4 can determine whether the anti-WTA antibody binding to bacteria and complement activation.
- complement C3 and C4 deposition mediated by anti-WTA antibodies in the mutations prepared in Examples ⁇ 1-1> to ⁇ 1-6> were examined by flow cytometry.
- Example ⁇ 1-1> adult serum obtained from healthy adult volunteers is described by Jung DJ et al. , J. Immunol., 189: 4951-4959, 2012] was treated with the strain M0107 prepared in Example ⁇ 1-1> to prepare a serum treated with Staphylococcus. Meanwhile, the mutant strains of Examples ⁇ 1-1> to ⁇ 1-6> were incubated at 37 ° C. for 12 hours until the 0D 600 value was 1 to 1.2, washed twice with 1 ml of PBS, and 70% The strain was fixed using ethane. The fixed strains were washed twice with culture buffer (10 mM Tris, 140 mM NaCl (H 7.4), 10 mM CaCl 2 and 13 ⁇ 4 BSA).
- the results are shown in g to r of FIG.
- the gray areas in FIGS. 4 g to r are the results of analysis of the deposition of C3 and C4 on Staphylococcus without anti-WTA IgG, and the black lines are the results of analysis of the deposition of C3 and C4 in the presence of anti-WTA antibodies. .
- staphylococci synthesizing ⁇ -GlcNAcylated WTA such as parent strains (g, m), AtarM (j, p) and Adit A mutant bacteria (k, q) C3 and C4 deposition was observed only in the phase.
- the results show that [beta] GkNAcylated residues of libri-phosphate in WTA are essential for anti-WTA mediated complement activation.
- Example 3 Confirming Complement-Mediated Opsonin Phagocytosis Induction of Mutant Strains Since anti-WTA antibodies induce C3 deposition in Staphylococcus aureus with ⁇ -GlcNAcylated residues of libri-phosphate in WTA, The strains were presumed to be easily fed by human polymorphonuclear neutrophil (PMN). To quantify Staphylococcus aureus predated by Plli, the number of bacteria predated by 100 human PMNs was measured for the mutant bacteria prepared in Examples ⁇ 1-1> to ⁇ 1-6>.
- PMN polymorphonuclear neutrophil
- Peripheral blood mononuclear cells were isolated from healthy donors using polymorphprep solution (Nycomed Pharm As, Torshov, Norway) and then subjected to trypan blue exclusion. PMN was obtained. Subsequently, the PMN suspension ' solution (1 ⁇ 5 ⁇ 10 5 cells, 35 til) was added to 5 ⁇ l of opsonized bacteria (3.7 ⁇ 10 6 CFU: infection multiplication ⁇ 25) and incubated at 37 ° C. for 60 minutes. It was. After the incubation, the number of bacteria predated by 100 Plia was counted under a fluorescence retardation microscope. The results of the experiment are shown in FIG. 6, and the experimental data were expressed as the mean square standard deviation after repeated three times ( ⁇ .05).
- Example ⁇ 4x1> purified WTA (10 yg) was separated by PAGE (27%) and visualized by silver staining.
- the gel subjected to the PAGE was transferred to the barrel using purified water
- the GlcNAc amount of the WTA of the mutant strain was measured as follows.
- Example ⁇ 4—1 WTA obtained in Example ⁇ 4—1 was acid hydrolyzed with 6N HC1 for 3 hours in KXTC under vacuum, followed by De Gubareff T ea /. , J. Biol. Chem. , 223: 377-388, 1956, and the content of the inorganic phosphate was measured.
- the GlcNAc content was measured according to the method described in Enghofer E et al., Carbohydrate research, 76: 233-238, 1979, and the GlcNAc / Pi ratio was calculated from the content. The results are shown in Table 2.
- each WTA (5 nmole phosphate) dissolved in 60 ⁇ l PBS (pH 7.5) was applied to F96 Cert, maxisorp immune plates (triplicate, Nunc) and adsorbed overnight at room temperature.
- the WTA-coated microfolates were incubated for 2 hours at 4T: with human serum dissolved in 50 ⁇ of complete agent (10 mMTris-HCKpH 7.4), 140 mM NaCl, and 1) BSA).
- H + L mouse monoclonal anti-human ⁇ IgG antibody
- HRP horseradish peroxidase
- anti-WTA IgG, anti-Zliar / WTA IgG, anti-l ar5 WTA from the serum The amount of IgG and anti-arv! / S rA IgG was measured.
- Example ⁇ 2-1> the WTA isolated in Example ⁇ 2-1> was dissolved in lxPBS at a concentration of 5 nmol phosphate / 50 ⁇ 1, and then the WTA was put in 50 ⁇ / well in a 96 well plate and stored at room temperature for 2 days. The well plates were coated. Blocking buffer (20 mMMTris 150 mM NaCl pH 7.4 and 1% BSA) was added to the WTA-coated 96 well plate at 200 ⁇ l per well and blocked for 2 hours at room temperature. After blocking, 200 ⁇ l of a washed complete layer solution (lOmMTris 140mMNaCl ⁇ 7.4) was added to the plate and washed at room temperature for 1 minute.
- Blocking buffer (20 mMMTris 150 mM NaCl pH 7.4 and 1% BSA) was added to the WTA-coated 96 well plate at 200 ⁇ l per well and blocked for 2 hours at room temperature. After blocking, 200 ⁇ l of a washed complete layer solution
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Immunology (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Communicable Diseases (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Oncology (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
Abstract
Description
Claims
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US14/399,225 US20150147328A1 (en) | 2012-05-07 | 2013-05-07 | Vaccine composition for preventing staphyllococcus aureus infection |
EP13788643.8A EP2848257A4 (en) | 2012-05-07 | 2013-05-07 | VACCINE COMPOSITION FOR PREVENTING STAPHYLOCOCCUS AUREUS INFECTION |
KR20147032987A KR20150024315A (ko) | 2012-05-07 | 2013-05-07 | 포도상구균 감염 예방용 백신 조성물 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201261643697P | 2012-05-07 | 2012-05-07 | |
US61/643,697 | 2012-05-07 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2013168965A2 true WO2013168965A2 (ko) | 2013-11-14 |
WO2013168965A3 WO2013168965A3 (ko) | 2014-01-16 |
Family
ID=49551416
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/KR2013/003957 WO2013168965A2 (ko) | 2012-05-07 | 2013-05-07 | 포도상구균 감염 예방용 백신 조성물 |
Country Status (4)
Country | Link |
---|---|
US (1) | US20150147328A1 (ko) |
EP (1) | EP2848257A4 (ko) |
KR (1) | KR20150024315A (ko) |
WO (1) | WO2013168965A2 (ko) |
Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014193722A1 (en) * | 2013-05-31 | 2014-12-04 | Genentech, Inc. | Anti-wall teichoic antibodies and conjugates |
WO2014194247A1 (en) * | 2013-05-31 | 2014-12-04 | Genentech, Inc. | Anti-wall teichoic antibodies and conjugates |
WO2015183041A1 (ko) * | 2014-05-29 | 2015-12-03 | 주식회사 녹십자 | 포도상구균 감염 질환의 예방 또는 치료용 조성물 |
WO2017010845A1 (ko) * | 2015-07-15 | 2017-01-19 | 재단법인 목암생명과학연구소 | 포도상구균 감염 질환의 예방 또는 치료용 조성물 |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9803002B2 (en) | 2013-05-31 | 2017-10-31 | Genentench, Inc. | Anti-wall teichoic antibodies and conjugates |
CA3000591A1 (en) | 2015-10-13 | 2017-04-20 | Sanofi Pasteur | Immunogenic compositions against s. aureus |
CA3012046C (en) | 2016-03-04 | 2020-11-10 | Genentech, Inc. | Process for the preparation of an antibody-rifamycin conjugate |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4450150A (en) | 1973-05-17 | 1984-05-22 | Arthur D. Little, Inc. | Biodegradable, implantable drug delivery depots, and method for preparing and using the same |
US4675381A (en) | 1983-07-01 | 1987-06-23 | Battelle Memorial Institute | Biodegradable polypeptide and its use for the gradual release of drugs |
US5417986A (en) | 1984-03-16 | 1995-05-23 | The United States Of America As Represented By The Secretary Of The Army | Vaccines against diseases caused by enteropathogenic organisms using antigens encapsulated within biodegradable-biocompatible microspheres |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR100644953B1 (ko) * | 1998-08-31 | 2006-11-10 | 인히비텍스, 인코포레이티드 | 다성분 백신 |
WO2004043405A2 (en) * | 2002-11-12 | 2004-05-27 | The Brigham And Women's Hospital, Inc. | Polysaccharide vaccine for staphylococcal infections |
AU2003298770A1 (en) * | 2002-12-02 | 2004-06-23 | Biosynexus Inc | Wall teichoic acid as a target for anti-staphylococcal therapies and vaccines |
KR20110124060A (ko) * | 2010-05-10 | 2011-11-16 | 부산대학교 산학협력단 | Wta를 유효성분으로 함유하는 백신 조성물 |
-
2013
- 2013-05-07 KR KR20147032987A patent/KR20150024315A/ko not_active Application Discontinuation
- 2013-05-07 US US14/399,225 patent/US20150147328A1/en not_active Abandoned
- 2013-05-07 WO PCT/KR2013/003957 patent/WO2013168965A2/ko active Application Filing
- 2013-05-07 EP EP13788643.8A patent/EP2848257A4/en not_active Withdrawn
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4450150A (en) | 1973-05-17 | 1984-05-22 | Arthur D. Little, Inc. | Biodegradable, implantable drug delivery depots, and method for preparing and using the same |
US4675381A (en) | 1983-07-01 | 1987-06-23 | Battelle Memorial Institute | Biodegradable polypeptide and its use for the gradual release of drugs |
US5417986A (en) | 1984-03-16 | 1995-05-23 | The United States Of America As Represented By The Secretary Of The Army | Vaccines against diseases caused by enteropathogenic organisms using antigens encapsulated within biodegradable-biocompatible microspheres |
Non-Patent Citations (22)
Title |
---|
BROWN S ET AL., PROC. NATL. ACAD. SCI. U S A, vol. 109, 2012, pages 18909 - 18914 |
DE GUBAREFF T ET AL., J. BIOL. CHEM., vol. 223, 1956, pages 377 - 388 |
ENGHOFER E ET AL., CARBOHYDRATE RESEARCH, vol. 76, 1979, pages 233 - 238 |
GRÜNDLING A ET AL., PROC. NATL. ACAD. SCI. USA, vol. 104, 2007, pages 8478 - 83 |
JUERGENS WG ET AL., J. EXP. MED., vol. 117, 1963, pages 925 - 935 |
JUNG DJ ET AL., J. IMMUNOL., vol. 189, 2012, pages 4951 - 4959 |
LOWY FD, THE NEW ENGLAND JOURNAL OF MEDICINE, vol. 339, 1998, pages 520 - 532 |
NAKAYAMA M ET AL., J. IMMUNOL., vol. 189, no. 12, 2012, pages 5903 - 5911 |
NATHENSON SG ET AL., J. BIOL. CHEM., vol. 237, 1962, pages 3839 - 3841 |
NOVICK RP ET AL., EMBO. J, vol. 12, 1993, pages 3967 - 3975 |
OKU Y ET AL., JOURNAL OF BACTERIOLOGY, vol. 191, 2009, pages 141 - 151 |
PARK KH ET AL., J. BIOL. CHEM., vol. 285, no. 35, 2010, pages 27167 - 2775 |
See also references of EP2848257A4 |
SHIRATSUCHI A ET AL., IMMUNOLOGY, vol. 129, 2010, pages 268 - 277 |
SWOBODA JG ET AL., CHEMBIOCHEM., vol. 11, 2010, pages 35 - 45 |
TAO L ET AL., GENE, vol. 120, 1992, pages 105 - 110 |
VAGNER V ET AL., MICROBIOLOGY (READING, ENGLAND), vol. 144, 1998, pages 3097 - 3104 |
WEIDENMAIER C ET AL., NATURE MEDICINE, vol. 10, 2004, pages 243 - 245 |
WEIDENMAIER C ET AL., NATURE REVIEWS, vol. 6, 2008, pages 276 - 287 |
WEIDENMAIER C ET AL., PLOS ONE, vol. 5, 2010, pages EL3227 |
XIA G ET AL., INT. J. MED. MICROBIOL., vol. 300, 2010, pages 148 - 154 |
XIA G ET AL., J. BIOL. CHEM., vol. 285, 2010, pages 13405 - 13415 |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2014193722A1 (en) * | 2013-05-31 | 2014-12-04 | Genentech, Inc. | Anti-wall teichoic antibodies and conjugates |
WO2014194247A1 (en) * | 2013-05-31 | 2014-12-04 | Genentech, Inc. | Anti-wall teichoic antibodies and conjugates |
US9895450B2 (en) | 2013-05-31 | 2018-02-20 | Genentech, Inc. | Anti-wall teichoic antibodies and conjugates |
EP3381939A1 (en) * | 2013-05-31 | 2018-10-03 | Genentech, Inc. | Anti-wall teichoic antibodies and conjugates |
WO2015183041A1 (ko) * | 2014-05-29 | 2015-12-03 | 주식회사 녹십자 | 포도상구균 감염 질환의 예방 또는 치료용 조성물 |
CN106413737A (zh) * | 2014-05-29 | 2017-02-15 | 株式会社绿十字 | 用于预防或治疗金黄色葡萄球菌感染的组合物 |
WO2017010845A1 (ko) * | 2015-07-15 | 2017-01-19 | 재단법인 목암생명과학연구소 | 포도상구균 감염 질환의 예방 또는 치료용 조성물 |
Also Published As
Publication number | Publication date |
---|---|
EP2848257A2 (en) | 2015-03-18 |
EP2848257A4 (en) | 2015-12-02 |
KR20150024315A (ko) | 2015-03-06 |
US20150147328A1 (en) | 2015-05-28 |
WO2013168965A3 (ko) | 2014-01-16 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP6771499B2 (ja) | 新規の多糖及びその使用 | |
JP6335102B2 (ja) | Staphylococcus感染に対する多糖類ワクチン | |
WO2013168965A2 (ko) | 포도상구균 감염 예방용 백신 조성물 | |
US11529405B2 (en) | MDR E. coli immunogen | |
JP2008179634A (ja) | グラム陽性細菌のリポタイコ酸に特異的なオプソニン性モノクローナルおよびキメラ抗体 | |
EP1791967B1 (en) | Methods and compositions relating to mannuronic acid specific binding peptides | |
WO2013096166A1 (en) | PSEUDOMONAS AERUGINOSA OprM EPITOPES FOR USE IN DIAGNOSTICS AND THERAPEUTICS | |
Lee et al. | Glycoepitopes of staphylococcal wall |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 13788643 Country of ref document: EP Kind code of ref document: A2 |
|
WWE | Wipo information: entry into national phase |
Ref document number: 14399225 Country of ref document: US |
|
ENP | Entry into the national phase |
Ref document number: 20147032987 Country of ref document: KR Kind code of ref document: A |
|
REEP | Request for entry into the european phase |
Ref document number: 2013788643 Country of ref document: EP |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2013788643 Country of ref document: EP |