WO2023186054A1 - Antibody specifically recognizing c5a and application of antibody - Google Patents

Antibody specifically recognizing c5a and application of antibody Download PDF

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Publication number
WO2023186054A1
WO2023186054A1 PCT/CN2023/085301 CN2023085301W WO2023186054A1 WO 2023186054 A1 WO2023186054 A1 WO 2023186054A1 CN 2023085301 W CN2023085301 W CN 2023085301W WO 2023186054 A1 WO2023186054 A1 WO 2023186054A1
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amino acid
acid sequence
sequence seq
seq
antibody
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PCT/CN2023/085301
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French (fr)
Chinese (zh)
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李忠
缪刘杨
吴飞林
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舒泰神（北京）生物制药股份有限公司
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Priority to CN202380009755.5A priority Critical patent/CN117203230A/en
Publication of WO2023186054A1 publication Critical patent/WO2023186054A1/en

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    • 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
    • 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
    • A61K39/12—Viral antigens
    • 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
    • A61K39/12—Viral antigens
    • A61K39/215—Coronaviridae, e.g. avian infectious bronchitis virus
    • 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
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/64—General methods for preparing the vector, for introducing it into the cell or for selecting the vector-containing host
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/10—Cells modified by introduction of foreign genetic material

Definitions

  • the present application relates to antibodies that specifically recognize complement component 5a (C5a) and their preparation methods and uses, including uses for the treatment of autoimmune diseases, inflammation, cancer, pain, respiratory and/or transplantation-related diseases.
  • C5a complement component 5a
  • C5a is an active peptide in allergic reactions and inflammation, which is formed during the complement cascade by cleavage of complement component C5 by C5 convertase.
  • C5a stimulates mast cell degranulation, tumor necrosis factor- ⁇ (TNF- ⁇ ) and histamine release, and also recruits phagocytes to sites of infection and inflammation by increasing the expression of adhesion molecules on the surface of endothelial cells (Mollnes, T.E. et al. Blood 2002, 100, 1869–1877; Riedemann, N.C. et al. Immunity 2003, 19, 193–202).
  • C5a In some cases of pathological stimulation, such as post-transplant allograft rejection and asthma, C5a also leads to increased vascular permeability (Gueler, F. et al. J. Am. Soc. Nephrol. 2008, 19, 2302–2312; Krug ,N.et al.Am.J.Respir.Crit.Care Med.2001,164,1841-1843; Khan,M.A.et al.Proc.Natl.Acad.Sci.USA 2013,110,6061–6066).
  • Several studies have discussed C5a levels in serum. In a study by Lechner et al., the level of C5a in the control group was 8.34+2.05 (ng/mL) (Lechner, J.
  • TGF- ⁇ transforming growth factor- ⁇
  • C5a is a potent pro-inflammatory molecule that binds to a classic G protein-coupled receptor (GPCR) C5aRI (CD88) and triggers the activation of pro-inflammatory signaling pathways (Li, R. et al. FASEB J. 2013, 27, 855 –864).
  • GPCR G protein-coupled receptor
  • C5aR is widely expressed on non-myeloid cells, such as umbilical vascular endothelial cells (HUVEC), murine dermis, liver, lung and renal proximal tubules (Monsinjon, T. et al. FASEB J. 2003, 17, 1003–1014; Gerard, C. et al. Annu. Rev. Immunol. 1994, 12, 775–808; Haviland, D. L.
  • Patent application WO2011063980 discloses the antibody INab308 (InflaRx) against human C5a
  • WO2012088247 discloses the C5a antibody MEDI-7814 (MedImmune)
  • US10450370 discloses the C5a antibody BNJ383 (Alexion).
  • an isolated anti-C5a antibody comprising: (i) a V H comprising HC-CDR1, HC-CDR2 and HC as shown in the amino acid sequence of SEQ ID NO:25 -CDR3; and VL , which comprises LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO:35; (ii) VH , which comprises the amino acid sequence SEQ ID NO V H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as shown in: 26; and V L comprising LC-CDR1, LC-CDR2 and V L as shown in amino acid sequence SEQ ID NO: 36 LC-CDR3; (iii) VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO:27; and VL comprising the amino acid sequence SEQ ID LC
  • an isolated anti-C5a antibody comprising: (i) V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising and V L ,
  • the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 20, or Variants of the V L , including substitutions of up to about 5 amino acids in its LC-CDRs;
  • V H the V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC- CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H whose HC-CDRs comprise substitutions of up to about 5 amino acids; and VL , said VL comprising: LC-CDR1, which includes the amino acid sequence
  • HC-CDR2 which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 8, or a variant of the V H , which HC-CDRs include up to about 5 Substitutions of amino acids; and V L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO: 13, LC-CDR2 comprising the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO: 17 Sequence SEQ ID NO: 21, or a variant of the V L containing up to about 5 amino acid substitutions in its LC-CDRs; (iv) V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9, or a variant of the V H , whose HC-CDRs include Substitutions of up to about 5 amino
  • LC-CDR2 which comprises the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , which contains up to about 5 LC-CDRs Substitution of amino acids; or (x) V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, and HC-CDR3, It comprises the amino acid sequence SEQ ID NO: 11, or a variant of said V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC-CDR1 comprising amino acids Sequence SEQ ID NO: 16, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 19, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 24, or a variant of the V L in which LC-CDRs Contains substitutions of up to about 5 amino acids.
  • any of the isolated anti-C5a antibodies as described above which includes: (i) VH , which includes the amino acid sequence shown in SEQ ID NO: 25 or a variant thereof, which variant is identical to SEQ
  • the amino acid sequence shown in ID NO:25 has at least about 80% sequence identity
  • VL which includes the amino acid sequence shown in SEQ ID NO:35 or a variant thereof, which variant is identical to the amino acid sequence shown in SEQ ID NO:35
  • the amino acid sequence shown has at least about 80% sequence identity
  • V H which includes the amino acid sequence shown in SEQ ID NO: 26 or a variant thereof, which variant is identical to the amino acid sequence shown in SEQ ID NO: 26
  • V L comprising the amino acid sequence set forth in SEQ ID NO: 36 or a variant thereof that has at least about 80% identity to the amino acid sequence set forth in SEQ ID NO: 36 % sequence identity
  • V H comprising the amino acid sequence set forth in SEQ ID NO:
  • the isolated anti-C5a antibody binds to human C5a with a Kd value of 0.1 pM to 1 nM.
  • an isolated anti-C5a antibody is provided that competes with any of the isolated anti-C5a antibodies described above for specific binding to C5a. In some embodiments, an isolated anti-C5a antibody is provided that specifically binds to the same epitope as any of the isolated anti-C5a antibodies described above.
  • any of the isolated anti-C5a antibodies described above, the isolated anti-C5a antibody comprises an Fc fragment.
  • the isolated anti-C5a antibody is a full-length IgG antibody.
  • the isolated anti-C5a antibody is a full-length IgG1 or IgG4 antibody.
  • the isolated anti-C5a antibody is a chimeric, fully human, or humanized antibody.
  • the isolated anti-C5a antibody is an antigen-binding fragment selected from the group consisting of Fab, Fab', F(ab)' 2 , Fab'-SH, single chain Fv (scFv), Fv In the group consisting of fragments, dAbs, Fds, nanobodies, diabodies and linear antibodies.
  • an isolated nucleic acid molecule encoding any of the anti-C5a antibodies described above is provided.
  • a vector is provided comprising any of the nucleic acid molecules described above.
  • a host cell is provided, the host cell comprising any one of the anti-C5a antibodies described above, any one of the nucleic acid molecules described above, or any one of the vectors described above.
  • a method for preparing an anti-C5a antibody is provided, which includes: a) culturing any of the above host cells under conditions that can effectively express the anti-C5a antibody; and b) obtaining the expressed anti-C5a antibody from the host cell. C5a antibody.
  • a method of treating a disease or condition in a desired individual comprising administering to the individual an effective amount of any of the anti-C5a antibodies described above.
  • provided is the use of any of the anti-C5a antibodies described above in the preparation of a pharmaceutical composition for treating a disease or condition in a desired individual.
  • the use of any of the anti-C5a antibodies described above or a pharmaceutical composition comprising an anti-C5a antibody in the preparation of a medicament for treating a disease or condition is provided.
  • the disease or disorder is related to the C5a signaling pathway, including autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplant immune dysregulation diseases or disorders .
  • the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
  • SIRS inflammatory response syndrome
  • a conjugate which includes a monoclonal antibody and a coupling moiety, wherein the monoclonal antibody is any anti-C5a antibody described herein, and the coupling moiety is a radioactive Isotopes, fluorescent substances, luminescent substances, colored substances or enzymes.
  • compositions, kits and manufactured products containing any of the anti-C5a antibodies mentioned above are also provided.
  • kits or article of manufacture includes any one of the anti-C5a antibodies or conjugates described herein.
  • any of the anti-C5a antibodies or conjugates described in this application is directed to the preparation of a kit or article of manufacture for detecting the presence or level of C5a in a sample.
  • an in vitro method of detecting the presence or level of C5a in a sample comprising: a) contacting the sample with any of the methods described herein under conditions that allow the formation of a complex between the antibody and C5a. contact an anti-C5a antibody or conjugate; b) detect the formation of a complex; and c) determine the presence or level of C5a in the sample.
  • a method for diagnosing abnormalities in C5a levels comprising: a) obtaining a sample to be tested, and b) combining the sample to be tested with the method described herein under conditions that allow the formation of a complex between the antibody and C5a. Contact any of the anti-C5a antibodies or conjugates; c) detect the formation of the complex; d) determine the level of C5a in the sample to be tested; and e) determine whether the level of C5a in the sample to be tested is higher than normal Levels of C5a in the sample.
  • the sample is a serum sample or plasma sample.
  • the sample to be tested is derived from a patient in need.
  • the patient has or has not received C5a antibody treatment.
  • Figures 1A-1C are the binding curves of anti-C5a antibodies RH23, RH25, RH32, RH35, RH36, RH58, RH85, RH86, RH92, RH95 and human C5a.
  • the application provides anti-C5a antibody molecules.
  • anti-C5a antibody molecules Through a combination of hybridoma technology, scFv phage library screening, and appropriately designed biochemical and biological experiments, highly efficient antibody molecules capable of binding human C5a and inhibiting the interaction of human C5a with its receptor have been identified.
  • the results presented here demonstrate that the antibodies in this application bind human C5a or human C5a-desArg with high affinity compared to the known anti-C5a antibody INab308.
  • Anti-C5a antibodies provided by this application include, for example, full-length anti-C5a antibodies, anti-C5a single chain antibodies (scFvs), anti-C5a Fc fusion proteins, multispecific (such as bispecific) anti-C5a antibodies, anti-C5a immunoconjugates Connected objects and the like.
  • the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 2, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 6, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 20, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
  • the application provides an anti-C5a antibody, the anti-C5a antibody comprising VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L Comprising: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 20, or the V L Variants containing up to about 5 amino acid substitutions in their LC-CDRs.
  • VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3,
  • the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 8, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which contains the amino acid sequence SEQ ID NO: 13, LC-CDR2, which contains the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which contains the amino acid sequence SEQ ID NO: 21, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
  • the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 9, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
  • the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 5, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 10, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
  • the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
  • the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 9, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 23, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
  • the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which contains the amino acid sequence SEQ ID NO: 15, LC-CDR2, which contains the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which contains the amino acid sequence SEQ ID NO: 22, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
  • the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
  • the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 11, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 16, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 19, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 24, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
  • nucleic acids encoding anti-C5a antibodies are also provided.
  • compositions containing anti-C5a antibodies are also provided.
  • treatment is a method of obtaining beneficial or desired results, including clinical results.
  • said beneficial or desired clinical results Including but not limited to one or more of the following: alleviating one or more symptoms caused by the disease, reducing the severity of the disease, stabilizing the disease (e.g., preventing or delaying the progression of the disease), preventing or delaying the spread of the disease (e.g., metastasis) , prevent or delay disease recurrence, delay or slow disease progression, improve disease status, alleviate disease (partial or complete), reduce the dose of one or more other drugs required to treat the disease, delay disease progression, improve or improve quality of life , increase body weight, and/or prolong survival.
  • treatment includes reduction of pathological consequences of disease (e.g., for cancer, tumor volume). The methods of the present application contemplate any one or more aspects of these treatments .
  • antibody includes full-length antibodies and antigen-binding fragments thereof.
  • Full-length antibodies include two heavy chains and two light chains.
  • the variable regions of the light and heavy chains are responsible for antigen binding.
  • the variable regions in the two chains usually include three hypervariable loops, called complementarity determining regions (CDRs) (light chain (LC) CDRs include LC-CDR1, LC-CDR2 and LC-CDR3, heavy chain (HC) )CDRs include HC-CDR1, HC-CDR2 and HC-CDR3).
  • CDRs complementarity determining regions
  • the CDR boundaries of the antibodies or antigen-binding fragments disclosed herein may be defined or identified by the Kabat, Chothia or Al-Lazikani conventions (Al-Lazikani 1997; Chothia 1985; Chothia 1987; Chothia 1989; Kabat 1987; Kabat 1991).
  • the three CDR regions of the heavy or light chain are inserted between flanking segments called framework regions (FRs), which are more conserved than the CDR regions and form a scaffold supporting the hypervariable loops.
  • FRs flanking segments
  • the constant regions of the heavy and light chains are not involved in antigen binding but exhibit a variety of effector functions.
  • Antibodies are classified based on the amino acid sequence of their heavy chain constant region.
  • the five major classes or isotypes of antibodies are IgA, IgD, IgE, IgG, and IgM, characterized by heavy chains of the alpha, delta, epsilon, gamma, and mu types, respectively.
  • major antibody classes are divided into subclasses, such as IgG1 ( ⁇ 1 heavy chain), IgG2 ( ⁇ 2 heavy chain), IgG3 ( ⁇ 3 heavy chain), IgG4 ( ⁇ 4 heavy chain), IgA1 ( ⁇ 1 heavy chain), or IgA2 ( ⁇ 2 heavy chain).
  • antigen-binding fragment includes antibody fragments, e.g., diabodies, Fab, Fab', F(ab') 2 , Fv fragments, disulfide-stabilized Fv fragments (dsFv), (dsFv) 2.
  • Bispecific dsFv (dsFv-dsFv'), disulfide bond-stabilized diabody (ds diabody), single-chain Fv (scFv), scFv dimer (bivalent diabody), Multispecific antibodies consisting of antibody fragments containing one or more CDRs, single domain antibodies, nanobodies, domain antibodies, bivalent domain antibodies, or any other antibody fragment capable of binding to an antigen but not containing a complete antibody structure .
  • the antigen-binding fragment is capable of binding to the same antigen as the parent antibody or parent antibody fragment (eg, parent scFv).
  • Antigen-binding fragments also include fusion proteins containing the above-described antibody fragments.
  • the antigen-binding fragment may include one or more CDRs from a specific human antibody grafted to framework regions from one or more different human antibodies.
  • epitope refers to a specific group of atoms or amino acids on an antigen to which an antibody or antibody portion binds. If two antibodies or antibody portions exhibit competitive binding to an antigen, they may bind to the same epitope on the antigen.
  • a first antibody inhibits binding of a second antibody to a C5a target by at least 50% (e.g., at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%) at equimolar concentrations , 95%, 98% or 99%), the first antibody "competes" with the second antibody for binding to the C5a target, and vice versa.
  • PCT publication WO 03/48731 describes a cross-competition based high-throughput antibody "epitope classification" approach.
  • the terms “specifically bind,” “specifically recognize,” or “specific for” refer to a measurable and reproducible interaction, e.g., binding of an antibody to a target can The presence of the target in a heterogeneous population of molecules, including biomolecules, is determined.
  • the ability of an antibody to specifically recognize a target means that the antibody binds to the target with higher affinity, avidity, easier and/or longer duration than to other targets.
  • an antibody that specifically recognizes an antigen reacts with one or more epitopes of the antigen with a binding affinity that is at least 10 times greater than its binding affinity to other targets.
  • an "isolated" anti-C5a antibody is an anti-C5a antibody that (1) is not related to a naturally occurring protein, (2) does not contain other proteins from the same source, and (3) is produced from a different species Expressed by cells of the genus, or (4) does not exist in nature.
  • isolated nucleic acid refers to nucleic acids of genomic, cDNA or synthetic origin, or combinations thereof. Depending on its source, the “isolated nucleic acid” refers to (1) not related to all or part of the polynucleotides in the "isolated nucleic acid” found in nature, (2) may not be related to polynucleotides that are not associated with it in the natural state. The nucleotides are operably linked, or (3) do not occur in nature as part of a longer sequence.
  • CDR complementarity determining region
  • chimeric antibody means that a portion of the heavy chain and/or light chain is identical or homologous to the corresponding sequence in an antibody from a specific species or belonging to a specific antibody class or subclass, and this chain(s) Antibodies whose remainder are identical to or homologous to corresponding sequences in antibodies from another genus or belonging to other antibody classes or subclasses, as well as fragments of such antibodies, as long as they have the biological activity in this application ( See U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)).
  • Fv is the smallest antibody fragment that contains complete antigen recognition and binding sites. This fragment is a dimer formed by a heavy chain variable domain and a light chain variable domain tightly non-covalently linked. The folding of these two domains results in 6 hypervariable loops (3 loops each in the light chain and heavy chain), which provide the antibody with the amino acid residues for binding to the antigen and confer a binding relationship between the antibody and the antigen. Binding specificity. However, even a single variable domain (or half of an Fv fragment, which contains only the 3 CDRs specific for the antigen) has the ability to recognize and bind the antigen, albeit with lower affinity than the complete binding site.
  • the scFv polypeptide further includes a linker polypeptide between the VH and VL domains that allows the scFv to form an ideal structure for antigen binding.
  • diabody is a small antibody fragment prepared by using a short linker (for example, 5 to 10 residues) between the V H and V L domains to construct an scFv fragment (see the previous paragraph). This allows the variable domains to pair inter-chain rather than intra-chain, producing a bivalent fragment, that is, a fragment with two antigen-binding sites.
  • Bispecific diabodies are heterodimers of two "cross-over" scFv fragments, in which the VH and VL domains of the two antibodies are located on different polypeptide chains.
  • Diabodies are fully described in EP 404,097; WO 93/11161; Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993).
  • Humanized forms of non-human (eg, rodent) antibodies are chimeric antibodies, which include minimal sequence from the non-human antibody.
  • humanized antibodies are human immunoglobulins (recipient antibodies) in which the hypervariable region (HVR) residues of the receptor antibody are modified from a non-human species such as mouse, rat, rabbit or Replacement of hypervariable region residues that are non-human mammalian and possess the desired antibody specificity, affinity and performance (donor antibody).
  • HVR hypervariable region residues of the receptor antibody
  • donor antibody residues in the human immunoglobulin framework region are replaced with corresponding non-human residues.
  • humanized antibodies may include residues that are not present in either the recipient antibody or the donor antibody. These modifications can further improve antibody performance.
  • a humanized antibody will contain substantially all, at least one, and usually two variable domains in which all or substantially all of the hypervariable loops correspond to hypervariable loops of a non-human immunoglobulin, and all or Essentially all framework regions are human immunoglobulin sequences.
  • the human antibody optionally also includes at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin.
  • Fc immunoglobulin constant region
  • Percent amino acid sequence identity or “homology” of the polypeptide and antibody sequences identified herein is defined as follows: Sequence comparisons are made where conservative substitutions are considered part of the sequence identity, and the candidate sequence is compared with the candidate sequence to be Compares the percentage of identical amino acid residues in a polypeptide sequence. Percent amino acid sequence identity can be determined by a variety of alignment methods within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, Megalign (DNASTAR), or MUSCLE software. One skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms required to maximize alignment over the full length of the sequences being compared.
  • Fc receptor or "FcR” is used to describe a receptor that binds to the Fc region of an antibody.
  • the FcR described herein is an FcR that binds an IgG antibody, a gamma receptor, including receptors of the Fc ⁇ RI, Fc ⁇ RII, and Fc ⁇ RIII subclasses, including allelic variants and agonist variants of these receptors. Change splicing form.
  • Fc ⁇ RII receptors include Fc ⁇ RIIA (“activating receptor”) and Fc ⁇ RIIB (“inhibitory receptor”), which have similar amino acid sequences and differ mainly in the cytoplasmic domain.
  • the cytoplasmic domain of activating receptor Fc ⁇ RIIA contains an immunoreceptor tyrosine activation motif (ITAM).
  • the cytoplasmic domain of the inhibitory receptor Fc ⁇ RIIB contains an immunoreceptor tyrosine inhibitory motif (ITIM) (see M.in Annu. Rev. Immunol. 15:203-234 (1997)).
  • ITAM immunoreceptor tyrosine activation motif
  • ITIM immunoreceptor tyrosine inhibitory motif
  • FcR in this application covers other types of FcRs, including FcRs identified in the future.
  • FcR also includes the neonatal receptor FcRn, which is responsible for the transfer of maternal IgGs to the neonate (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994) )).
  • FcRn refers to the neonatal Fc receptor (FcRn).
  • FcRn is structurally similar to the major histocompatibility complex (MHC) and consists of an ⁇ chain non-covalently bound to ⁇ 2 microglobulin.
  • MHC major histocompatibility complex
  • FcRn plays an important role in the passive transport of immunoglobulin IgGs from mother to newborn and in regulating serum IgG levels.
  • FcRn serves as a rescue receptor that binds and transports endocytosed IgG in an intact form within and between cells and protects them from default degradation pathways.
  • the "CH1 domain" of the human IgG heavy chain constant region generally extends from amino acid 118 to amino acid 215 (EU numbering system).
  • the "hinge region” is generally defined as extending from position Glu 216 to position Pro 230 of human IgG1 (Burton, Molec. Immunol. 22:161-206 (1985)). By placing the first and last cysteine residues that form the inter-heavy chain disulfide bond after the same position as IgG1, the hinge regions of other IgG isotypes can be aligned with the IgG1 sequence.
  • the "CH2 domain" of the human IgG Fc region usually extends from amino acid 231 to amino acid 340.
  • the unique feature of the CH2 domain is that it does not pair closely with another region, but instead has two N-terminal linked branched sugar chains inserted between the two CH2 domains of the intact natural IgG molecule. It is speculated that sugars may serve as a surrogate for domain-to-domain pairing, helping to keep the CH2 domain stable.
  • the "CH3" domain includes the region extending from the C-terminal residues within the Fc region to the CH2 domain (from amino acid 341 to the C-terminus of the antibody sequence, usually amino acid residue 446 or 447 of IgG).
  • a “functional Fc fragment” has the "effector function” possessed by a native Fc region sequence.
  • exemplary “effector functions” include Clq binding; complement-dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors ( Such as B cell receptor; BCR), etc.
  • effector functions typically require the binding of an Fc region to a binding domain (eg, an antibody variable region) and can be assessed using a variety of experimental methods well known in the art.
  • Antibodies with IgG Fc variants that have "altered" FcR binding affinity or ADCC activity have increased or decreased FcR binding activity and/or ADCC activity compared to the parent polypeptide or a polypeptide comprising a native Fc sequence.
  • An Fc variant that exhibits "enhanced binding" to an FcR has a higher binding affinity (e.g., a lower apparent Kd or IC50 value) to at least one FcR compared to the parent polypeptide or a polypeptide comprising a native IgG Fc sequence. ).
  • the binding capacity is enhanced 3-fold, such as 5, 10, 25, 50, 60, 100, 150, 200, even up to 500-fold or a 25% to 1000% increase in binding capacity compared to the parent polypeptide.
  • An Fc variant exhibits "reduced binding" to an FcR, having a lower affinity (eg, a higher apparent Kd or IC50 value) for at least one FcR compared to the parent polypeptide. Its binding capacity is reduced by 40% or more compared to the parent polypeptide.
  • ADCC antibody-dependent cell-mediated cytotoxicity
  • cytotoxic cells e.g., natural killer cells (NK), neutrophils
  • FcRs Fc receptors
  • Antibodies “arm” cytotoxic cells and are required for this killing.
  • NK cells only express Fc ⁇ RIII
  • monocytes express Fc ⁇ RI, Fc ⁇ RII, and Fc ⁇ RIII.
  • FcR FcR on hematopoietic cells
  • Table 3 The expression of FcR on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991).
  • an in vitro ADCC assay can be performed, as described in U.S. Patent No. 5,500,362 or 5,821,337. Effector cells suitable for such experiments include peripheral blood mononuclear cells (PBMC) and natural killer cells (NK).
  • PBMC peripheral blood mononuclear cells
  • NK natural killer cells
  • the ADCC activity of the target molecule can also be assessed in vivo, for example in an animal model as disclosed in Clynes et al. PNAS (USA) 95:652-656 (1998).
  • the antibody polypeptide is essentially the same in quantity as the wild-type IgG Fc polypeptide (or parent polypeptide) during the experiment, it can more effectively mediate ADCC both in vitro and in vivo.
  • Such variants are generally identified using any in vitro ADCC assay known in the art, such as assays or methods for identifying ADCC activity, such as in animal models and the like. In some embodiments, such variants mediate ADCC 5- to 100-fold, for example, 25- to 50-fold more efficiently than wild-type Fc (or the parent polypeptide).
  • “Complement-dependent cytotoxicity” or “CDC” refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (C1q) to antibodies (subclasses with appropriate structures) that bind cognate antigens.
  • C1q the first component of the complement system
  • CDC experiments can be performed as described in Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996). Polypeptide variants with altered Fc region amino acid sequences and increased or decreased Clq binding capacity are described in US Patent No. 6,194,551 Bl and WO99/51642. The contents of these patent publications are expressly incorporated herein by reference. See also Idusogie et al. J. Immunol. 164:4178-4184 (2000).
  • nucleotide sequence encoding an amino acid sequence includes all nucleotide sequences that are degenerates of each other and encode the same amino acid sequence. Nucleotide sequences encoding proteins or RNA may also include introns, for example, nucleotide sequences encoding proteins may include introns in some forms.
  • operably linked refers to a functional linkage between a regulatory sequence and a heterologous nucleotide sequence, thereby allowing expression of the latter.
  • a first nucleotide sequence is operably linked to a second nucleotide sequence when the first nucleotide sequence is in a functional relationship with the second nucleotide sequence.
  • a promoter is operably linked to a coding sequence if it affects the transcription or expression of the coding sequence.
  • operably linked DNA sequences are contiguous and, if necessary, two protein-coding regions can be joined in the same reading frame.
  • “Homology” refers to sequence similarity or sequence identity between two polypeptides or between two nucleic acid molecules. If the same position of two compared sequences contains the same base or amino acid monomer subunit, for example, the same position of two DNA molecules both contains adenine, then the two DNA molecules are homologous at that position.
  • the percent homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the total number of positions multiplied by 100. For example, if 6 out of 10 positions in two sequences match or are homologous, the two sequences are 60% homologous. For example, the DNA sequences ATTGCC and TATGGC have 50% homology. Generally speaking, when comparing two sequences, the comparison is performed with the purpose of obtaining maximum homology.
  • an “effective amount” of an anti-C5a antibody or composition disclosed herein is an amount sufficient to achieve a specified purpose.
  • An “effective amount” can be determined empirically and by methods known to be relevant to the stated purpose.
  • a therapeutically effective amount refers to an amount of an anti-C5a antibody or composition thereof disclosed herein that is effective in treating a disease or condition in an individual.
  • a therapeutically effective amount of an anti-C5a antibody or composition thereof is one that reduces the number of cancer cells; reduces the size or weight of the tumor; inhibits (i.e., slows or preferably stops to a certain extent) the impact of tumor cells on surrounding cells. Invasion of organs; inhibition (i.e., slowing down or preferably stopping to a certain extent) tumor metastasis; inhibiting tumor growth to a certain extent, and/or alleviating one or more symptoms associated with cancer to a certain extent.
  • a therapeutically effective amount refers to an amount capable of prolonging the survival of a patient. In some embodiments, a therapeutically effective amount refers to an amount capable of improving progression-free survival of a patient.
  • pharmaceutically acceptable refers to a material that has no biological activity or other undesirable properties, e.g., the material can be incorporated into a pharmaceutical composition administered to a patient, and Does not cause significant adverse biological reactions or interact in a deleterious manner with any other components contained in the composition.
  • Pharmaceutically acceptable carriers or excipients preferably meet required standards for toxicological or manufacturing testing and/or are included in the Inactive Ingredient Guidelines prepared by the U.S. Food and Drug Administration.
  • Embodiments of the application described herein should be understood to include embodiments “consisting of” and/or “consisting essentially of.”
  • references herein to "about” a value or parameter include (and describe) variations on the value or parameter itself. For example, descriptions referring to “about X” include descriptions of "X”.
  • reference to "not" a value or parameter generally means and describes “other than” a value or parameter.
  • the method cannot be used to treat type X cancer, meaning the method is typically used to treat other types of cancer besides type X cancer.
  • the application provides anti-C5a antibodies that specifically bind C5a.
  • anti-C5a antibodies include, but are not limited to, humanized antibodies, chimeric antibodies, mouse antibodies, human antibodies, and antibody molecules comprising heavy chain and/or light chain CDRs as described herein.
  • the application provides isolated antibodies that bind C5a.
  • Contemplated anti-C5a antibodies include, for example, full-length anti-C5a antibodies (e.g., full-length IgG1 or IgG4), anti-C5a single chain antibodies, anti-C5a Fc fusion proteins, multispecific (e.g., bispecific) anti-C5a antibodies, anti-C5a Immunoconjugates, and the like.
  • the anti-C5a antibody is a full-length antibody (eg, full-length IgG1 or IgG4) or an antigen-binding fragment thereof that specifically binds C5a.
  • the anti-C5a antibody is Fab, Fab', F(ab)' 2 , Fab'-SH, single chain Fv (scFv), Fv fragment, dAb, Fd, nanobody, diabody (diabody) or linear antibody.
  • an antibody that specifically binds to C5a means that the affinity of the antibody for binding to C5a is at least 10 times greater than the binding affinity for the non-target (including, for example, 10, 10 2 , 10 3 , 10 4 , 10 5 , 10 6 , or 10 7 times).
  • non-target refers to an antigen that is not C5a.
  • Binding affinity can be determined by methods known in the art, such as ELISA, fluorescence-activated cell sorting (FACS) analysis or radioimmunoprecipitation analysis (RIA).
  • the Kd value can be determined by methods known in the art, such as surface plasmon resonance (SPR) technology or biolayer interference (BLI) technology.
  • non-human anti-C5a antibodies comprising human sequences are discussed broadly herein (eg, human heavy and light chain variable domains comprising human CDR sequences), non-human anti-C5a antibodies are also contemplated.
  • non-human anti-C5a antibodies include the human CDR sequences and non-human framework sequences of the anti-C5a antibodies described herein.
  • the non-human framework sequences include any for use as described herein.
  • non-human anti-C5a antibodies include anti-C5a antibodies generated by grafting one or more human CDR sequences described herein into non-human framework regions (eg, murine or chicken framework region sequences).
  • the complete amino acid sequence of an exemplary human C5a includes or consists of the amino acid sequence shown in SEQ ID NO: 49.
  • the anti-C5a antibodies described herein specifically recognize an epitope in human C5a. In some embodiments, the anti-C5a antibody cross-reacts with C5a of species other than human. In some embodiments, the anti-C5a antibody is fully specific for human C5a and does not cross-react with other non-human species.
  • the anti-C5a antibody cross-reacts with at least one allelic variant of the C5a protein (or fragment thereof).
  • the allelic variant has up to 30 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) compared to the naturally occurring C5a protein (or fragment thereof). , 15, 20, 25 or 30) amino acid substitutions (eg, conservative substitutions).
  • the anti-C5a antibody does not cross-react with any allelic variant of the C5a protein (or fragment thereof).
  • the anti-C5a antibody cross-reacts with at least one interspecies variant of the C5a protein.
  • the C5a protein (or fragment thereof) is human C5a
  • the interspecies variant of the C5a protein (or fragment thereof) is a variant in cynomolgus monkey.
  • the anti-C5a antibody does not cross-react with any interspecies variant of the C5a protein.
  • the anti-C5a antibody comprises an antibody heavy chain constant region and an antibody light chain constant region.
  • the anti-C5a antibody comprises an IgGl type heavy chain constant region.
  • the anti-C5a antibody comprises an IgG2-type heavy chain constant region.
  • the anti-C5a antibody comprises an IgG3 heavy chain constant region.
  • the anti-C5a antibody comprises an IgG4 type heavy chain constant region.
  • the heavy chain constant region comprises (including consists of or consists essentially of) the amino acid sequence SEQ ID NO: 45.
  • the heavy chain constant region comprises (including consists of or consists essentially of) the amino acid sequence SEQ ID NO: 46.
  • the anti-C5a antibody comprises a kappa light chain constant region.
  • the light chain constant region comprises (including consists of or consists essentially of) the amino acid sequence SEQ ID NO: 47.
  • the anti-C5a antibody comprises a lambda light chain constant region.
  • the light chain constant region comprises (including consists of or consists essentially of) the amino acid sequence SEQ ID NO: 48.
  • the anti-C5a antibody comprises an antibody heavy chain variable domain and an antibody light chain variable domain.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 6, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 20, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
  • the anti-C5a antibody comprises a VH comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:6; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:20.
  • the anti-C5a antibody comprises a V H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in the amino acid sequence of SEQ ID NO: 25; and a V L comprising VL as shown in the amino acid sequence SEQ ID NO:35 includes LC-CDR1, LC-CDR2 and LC-CDR3.
  • the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 25 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 25 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 35 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 35.
  • the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:25, and a VL comprising the amino acid sequence SEQ ID NO:35.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, HC -CDR3, which comprises the amino acid sequence SEQ ID NO:7, or a variant of said VH , whose HC-CDRs comprise up to about 5 amino acid substitutions; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 20, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
  • the anti-C5a antibody comprises a VH comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:7; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:20.
  • the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 as shown in the amino acid sequence of SEQ ID NO:26; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO:36 includes LC-CDR1, LC-CDR2 and LC-CDR3.
  • the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 26 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 26 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 36 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 36.
  • the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:26, and a VL comprising the amino acid sequence SEQ ID NO:36.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 8, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 13, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 21, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:8; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:13, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:21.
  • the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 as shown in the amino acid sequence of SEQ ID NO:27; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO:37 includes LC-CDR1, LC-CDR2 and LC-CDR3.
  • the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 27 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 27 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 37 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 37.
  • the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:27, and a VL comprising the amino acid sequence SEQ ID NO:37.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 9, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:9; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:22.
  • the anti-C5a antibody comprises a V H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as shown in the amino acid sequence of SEQ ID NO : 28; and a V L comprising VL as shown in the amino acid sequence SEQ ID NO:38 contains LC-CDR1, LC-CDR2 and LC-CDR3.
  • the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 28 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 28 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 38 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 38.
  • the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:28, and a VL comprising the amino acid sequence SEQ ID NO:38.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 5, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 10, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 5, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 10; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO: 14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:22.
  • the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 as shown in the amino acid sequence of SEQ ID NO:29; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO:39 includes LC-CDR1, LC-CDR2 and LC-CDR3.
  • the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 29 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 29 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 39 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 39.
  • the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:29, and a VL comprising the amino acid sequence SEQ ID NO:39.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
  • the anti-C5a antibody comprises a VH comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:7; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:22.
  • the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 as shown in the amino acid sequence of SEQ ID NO:30; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO:40 includes LC-CDR1, LC-CDR2 and LC-CDR3.
  • the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 30 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 30 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 40 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 40.
  • the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:30, and a VL comprising the amino acid sequence SEQ ID NO:40.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 9, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 23, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:9; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:23.
  • the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in the amino acid sequence SEQ ID NO:31 of VH ; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO: 41 includes LC-CDR1, LC-CDR2 and LC-CDR3.
  • the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 31 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 31 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 41 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 41.
  • the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:31, and a VL comprising the amino acid sequence SEQ ID NO:41.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, HC -CDR3, which comprises the amino acid sequence SEQ ID NO:7, or a variant of said VH , whose HC-CDRs comprise up to about 5 amino acid substitutions; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 15, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
  • the anti-C5a antibody comprises a VH comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:7; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:15, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:22.
  • the anti-C5a antibody comprises a V H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in the amino acid sequence of SEQ ID NO: 32; and a V L comprising VL as shown in the amino acid sequence SEQ ID NO: 42 includes LC-CDR1, LC-CDR2 and LC-CDR3.
  • the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 32 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 32 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and V L comprising the amino acid sequence SEQ ID NO: 42 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 42.
  • the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:32, and a VL comprising the amino acid sequence SEQ ID NO:42.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, HC -CDR3, which comprises the amino acid sequence SEQ ID NO:7, or a variant of said VH , whose HC-CDRs comprise up to about 5 amino acid substitutions; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:7; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:22.
  • the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in the amino acid sequence SEQ ID NO:33 of VH ; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO: 43 includes LC-CDR1, LC-CDR2 and LC-CDR3.
  • the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 33 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 33 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 43 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 43.
  • the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:33, and a VL comprising the amino acid sequence SEQ ID NO:43.
  • the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 11, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 16, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 19, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 24, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
  • the anti-C5a antibody comprises a VH comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 11; and VL, which VL comprises : LC-CDR1, which comprises the amino acid sequence SEQ ID NO: 16, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 19, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:24.
  • the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 as shown in the amino acid sequence of SEQ ID NO:34; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO: 44 includes LC-CDR1, LC-CDR2 and LC-CDR3.
  • the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 34 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 34 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 44 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 44.
  • the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:34, and a VL comprising the amino acid sequence SEQ ID NO:44.
  • amino acid substitutions are limited to the "exemplary substitutions” set forth in Table 4 herein. In some embodiments, amino acid substitutions are limited to the "preferred substitutions” set forth in Table 4 herein.
  • functional epitopes can be resolved by combined alanine scanning methods.
  • combinatorial alanine scanning techniques can be used to identify amino acids in the C5a protein that are necessary for interaction with anti-C5a antibodies.
  • the epitope is conformational and the crystal structure of an anti-C5a antibody bound to the C5a protein can be used to identify the epitope.
  • the application provides antibodies that compete for binding to C5a with any of the anti-C5a antibodies described herein.
  • antibodies are provided that are capable of binding to an epitope on C5a competitively with any of the anti-C5a antibodies described herein.
  • an anti-C5a antibody is provided that binds to the same epitope as an anti-C5a antibody molecule comprising VH and VL , wherein the VH comprises the amino acid set forth in any of SEQ ID NOs: 25-34 sequence, and the V L comprises the amino acid sequence shown in any one of SEQ ID NOs: 35-44.
  • an anti-C5a antibody that competes for binding to C5a with an anti-C5a antibody comprising VH and VL , wherein the VH comprises the amino acid sequence set forth in any one of SEQ ID NOs: 25-34 , and the V L includes the amino acid sequence shown in any one of SEQ ID NOs: 35-44.
  • competition experiments can be used to identify monoclonal antibodies that compete with the anti-C5a antibodies described herein for binding to C5a. Competition experiments can determine whether two antibodies bind the same epitope by identifying the same or spatially overlapping epitope or by one antibody competitively inhibiting the binding of another antibody to the antigen. In certain embodiments, such competing antibodies bind the same epitope as the antibodies described herein.
  • Some exemplary competition experiments include, but are not limited to, conventional experiments as mentioned in Harlow and Lane (1988) Antibodies: A Laboratory Manual ch.14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.).
  • each antibody is said to bind the same epitope if it blocks 50% or more of the binding of the other antibody.
  • the antibody that competes with an anti-C5a antibody described herein is a chimeric antibody, a humanized antibody, or a fully human antibody.
  • Exemplary anti-C5a antibody sequences are shown in Table 2 and Table 3, where CDR numbering is performed according to Kabat definition. Those skilled in the art will recognize that there are a variety of known algorithms for predicting the location of CDRs and defining antibody light and heavy chain variable regions. Antibodies containing the CDRs, VH and/or VL sequences of antibodies as described herein, but based on prediction algorithms other than those exemplified in the table below are also within the scope of the present application.
  • Complement component 5a (C5a) was first described as a cleavage product of complement factor 5 (C5) with chemotactic and anaphylatoxin properties (Shin et al., 1968).
  • the precursor of C5a, C5 protein contains 1676 amino acids, has a molecular weight of 188 kDa, and its gene is located at 9q33–9q34 (Wetsel et al., 1988).
  • Human C5a is an approximately 11 kDa glycoprotein containing 74 amino acids produced by C5 convertase cleavage of the ⁇ chain of C5, but N-linked glycosylation is not essential for its function.
  • the properties of C5a suggest that it is an important component of the innate immune response, but existing evidence suggests that C5a may also play a role in adaptive immunity (Kohl, 2006).
  • C5a is not necessarily a trigger of inflammatory diseases, excessive or uncontrolled C5a is produced in many inflammatory diseases, suggesting that C5a can promote and maintain inflammatory responses (Guo and Ward, 2005).
  • C5a has four antiparallel ⁇ -helices connected by peptide loops and is made more stable by three key disulfide bonds (Monk et al., 2007). Mutagenesis and antibody studies have identified several essential residues that provide interaction with the receptor (reviewed by Monk et al., 2007).
  • the complement cascade can be activated through four pathways: the classical pathway, the alternative pathway, the mannan-binding lectin pathway (MBL) or the exogenous protease pathway (Ricklin and Lambris, 2007).
  • the classical pathway and the lectin pathway are activated by recognition of antibody complexes formed on the surface of pathogens and mannose on bacterial surfaces, respectively. Both pathways result in cleavage of C4 by serine proteases, forming C4a and C4b. After C4b binds to C2, it leads to the generation of C2a under the action of protease. C4b and C2a form the C3 convertase (C4b2a) in the classical pathway.
  • the alternative pathway can be activated by foreign body surfaces or "slow transport", resulting in spontaneous hydrolysis of C3, which subsequently binds to factor B and forms C3 convertase (C3bBb) in the alternative pathway, which continues to maintain low levels Activation of the complement cascade, thereby ensuring a rapid response to invading pathogens (Ricklin and Lambris, 2007).
  • the above three pathways can form C3 convertase, and C3 convertase further cleaves the C3 protein to form C3a and C3b.
  • C3b can promote cell recognition of pathogen surfaces and clearance of pathogens, and can also form C5 convertase (C4b2aC3b or C3bBbC3b) with C3 convertase (C4b2a or C3bBb), and then C5 convertase further cleaves C5 to produce C5a and C5b.
  • C5b proceeds to initiate the assembly of the membrane attack complex (MAC; C5b-9).
  • the complement cascade is tightly regulated by a series of soluble membrane-bound regulatory proteins that prevent complement activation products from targeting host tissues (Ricklin and Lambris, 2007). However, this control can be circumvented through some exogenous pathways.
  • thrombin can directly cleave C3 and C5, causing activation of the complement system (Amara et al., 2008).
  • activated neutrophils and alveolar macrophages can cleave C5 to produce C5a through secreted serine proteases (Amara et al., 2008).
  • Carboxypeptidase on the plasma and cell surface can remove the arginine at the C-terminus of the protein, so the C5a produced by C5 cleavage can be rapidly metabolized by it to form C5adesArg (Bokisch and Muller-Eberhard, 1970).
  • C5adesArg Compared with C5a, C5adesArg has reduced potency, resulting in reduced binding affinity to the classical C5a receptor CD88 (Higginbottom et al., 2005). C5a and C5adesArg can be cleared rapidly in the body. About 50% of C5a and C5adesArg are cleared from the circulation within 2-3 minutes. In addition, some C5a is mediated after binding to CD88 on leukocytes and other cells (Oppermann and Gotze, 1994). However, a second receptor, C5a-like receptor 2 (C5L2), can more efficiently remove complement fragments by rapidly internalizing C5a/C5adesArg and, in particular, C5adesArg, allowing it to be retained and degraded in certain cell types (Scola (2009). In contrast, CD88-expressing cells internalize C5a and release a higher proportion of C5a in an undegraded, presumably active form. Plasma C5a can also be cleared by the liver (Chenoweth and Goodman , 1983).
  • C5a binds CD88 and C5L2 with similarly high affinity.
  • the affinity of C5adesArg for C5L2 is similar to that of C5a ( 12nM), but has a much lower affinity for CD88 ( 660 nM) (Monk et al., 2007).
  • CD88 and C5L2 share 35% sequence homology and are located in the same region of chromosome 19 (19q13.3–19q13.4). They cluster with other chemokine receptor genes such as the formyl peptide receptor family and bradykinin receptors. Both are glycosylated seven-transmembrane proteins with a molecular weight of approximately 45 kDa.
  • CD88 is a G protein-coupled receptor and a member of the rhodopsin gene family (Monk et al., 2007). Binding of C5a to CD88 is thought to occur at two distinct and physically separate sites. The first "recognition” site is located at the extracellular amino terminus (N-terminus) of the receptor, which binds to the C5a N-terminus and disulfide-linked core. The second “activation” site is formed by the transmembrane domain of the receptor, which binds to the C-terminus of C5a and leads to the generation of a specific signaling pathway mediated by the receptor-coupled G protein (Monk et al., 2007 ).
  • C5L2 Cell types that express C5L2 are broadly the same as those that express CD88, such as neutrophils, monocytes, lymphocytes, macrophages, as well as non-myeloid cells such as vascular smooth muscle cells and cells of tissue origin such as the adrenal gland. , heart, liver, lung, spleen and brain), but under non-inflammatory conditions, the expression level of C5L2 is significantly lower than that of CD88 (Gao et al., 2005). The function of C5L2 is unknown. Some experimental data suggest that C5L2 may serve as a decoy receptor without signal transduction function.
  • C5L2 has anti-inflammatory functions, possibly by reducing the amount of C5a that can bind to CD88.
  • C5L2 can act as a positive regulator that is critical for C5a and C3a signaling, at least in mice (Chen et al., 2007).
  • C5L2 was found to be critical for promoting C5a signaling in neutrophils, macrophages, and fibroblasts, whereas lack of C5L2 in vivo resulted in ovalbumin-induced airway hyperresponsiveness and inflammation (Chen et al. , 2007).
  • a mouse model of “late” sepsis (100% lethality) only simultaneous blockade of C5L2 and CD88 was protective (Rittirsch et al., 2008).
  • the anti-C5a antibody is a full-length anti-C5a antibody.
  • the full-length anti-C5a antibody is IgA, IgD, IgE, IgG, or IgM.
  • the full-length anti-C5a antibody comprises an IgG constant region, such as that of IgG1, IgG2, IgG3, IgG4, or a variant thereof.
  • the full-length anti-C5a antibody comprises a lambda light chain constant region.
  • the full-length anti-C5a antibody comprises a kappa light chain constant region.
  • the full-length anti-C5a antibody is a full-length human anti-C5a antibody. In some embodiments, the full-length anti-C5a antibody comprises mouse immunoglobulin Fc sequence. In some embodiments, the full-length anti-C5a antibody comprises an Fc sequence that has been altered or otherwise altered such that it has enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity. Effector functions of (CDC).
  • ADCC antibody-dependent cell-mediated cytotoxicity
  • CDC complement-dependent cytotoxicity
  • a full-length anti-C5a antibody comprising an IgG1 constant region that specifically binds to C5a.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG2 constant region is provided, the anti-C5a antibody specifically binding to C5a.
  • the IgG2 is human IgG2.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a full-length anti-C5a antibody comprising an IgG3 constant region is provided, the anti-C5a antibody specifically binding to C5a.
  • the IgG3 is human IgG3.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region is provided, the anti-C5a antibody specifically binding to C5a.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO: 1 or a variant thereof, which variant includes substitutions of up to about 3 (e.g., 1, 2 or 3) amino acids; HC-CDR2, which includes SEQ ID NOs: The amino acid sequence shown in any one of 2-5, or a variant thereof, the variant comprising a substitution of up to about 3 (e.g., 1, 2 or 3) amino acids; and HC-CDR3, which comprises SEQ ID NOs: 6 - the amino acid sequence shown in any one of 11 or a variant thereof, the variant comprising a substitution of up to about 3 (eg 1, 2 or 3) amino acids; and b) a light chain variable domain, the light chain variable domain
  • the chain variable domain includes: LC-CDR1, which includes the
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG2 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO: 1 or a variant thereof, which variant includes substitutions of up to about 3 (e.g., 1, 2 or 3) amino acids; HC-CDR2, which includes SEQ ID NOs: The amino acid sequence shown in any one of 2-5, or a variant thereof, the variant comprising a substitution of up to about 3 (e.g., 1, 2 or 3) amino acids; and HC-CDR3, which comprises SEQ ID NOs: 6 - the amino acid sequence shown in any one of 11 or a variant thereof, the variant comprising a substitution of up to about 3 (eg 1, 2 or 3) amino acids; and b) a light chain variable domain, the light chain variable domain
  • the chain variable domain includes: LC-CDR1, which includes the
  • the IgG2 is human IgG2.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a full-length anti-C5a antibody comprising an IgG3 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO: 1 or a variant thereof, which variant includes a substitution of up to about 3 (e.g., 1, 2 or 3) amino acids, HC-CDR2, which includes SEQ ID NOs: The amino acid sequence shown in any one of 2-5, or a variant thereof, the variant comprising a substitution of up to about 3 (e.g., 1, 2 or 3) amino acids; and HC-CDR3, which comprises SEQ ID NOs: 6 - the amino acid sequence shown in any one of 11 or a variant thereof, the variant comprising a substitution of up to about 3 (eg 1, 2 or 3) amino acids; and b) a light chain variable domain, the light chain variable domain
  • the chain variable domain includes: LC-CDR1, which includes
  • the IgG3 is human IgG3.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO: 1 or a variant thereof, which variant includes substitutions of up to about 3 (e.g., 1, 2 or 3) amino acids; HC-CDR2, which includes SEQ ID NOs: The amino acid sequence shown in any one of 2-5, or a variant thereof, the variant comprising a substitution of up to about 3 (e.g., 1, 2 or 3) amino acids; and HC-CDR3, which comprises SEQ ID NOs: 6 - the amino acid sequence shown in any one of 11 or a variant thereof, the variant comprising a substitution of up to about 3 (eg 1, 2 or 3) amino acids; and b) a light chain variable domain, the light chain variable domain
  • the chain variable domain includes: LC-CDR1, which includes the
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO:1, HC-CDR2, which includes the amino acid sequence shown in any one of SEQ ID NOs:2-5, and HC-CDR3, which includes SEQ ID NOs:6-11 The amino acid sequence shown in any one of them, or a variant of the heavy chain variable domain, the HC-CDR sequence of which contains at most about 5 (such as 1, 2, 3, 4 or 5) amino acid substitutions; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs: 12-16, LC-CDR2 comprising SEQ ID The amino acid sequence shown in any one of NOs: 17-19, and LC-CDR3, which includes the amino acid sequence shown in any one
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region wherein the anti-C5a antibody comprises a) a heavy chain variable domain comprising: HC-CDR1, It includes the amino acid sequence shown in SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence shown in any of SEQ ID NOs: 2-5, and HC-CDR3, which includes the amino acid sequence shown in any of SEQ ID NOs: 6-11 Any of the amino acid sequences shown, or a variant of the heavy chain variable domain, comprising up to about 5 (e.g., 1, 2, 3, 4 or 5) amino acid substitutions in the HC-CDR sequence; and b) Light chain variable domain, the light chain variable domain comprising: LC-CDR1, comprising the amino acid sequence shown in any one of SEQ ID NOs: 12-16, LC-CDR2, comprising SEQ ID NOs: The amino acid sequence shown in any one of 17-19, and LC-CDR3, which includes the amino acid sequence shown
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO:1, HC-CDR2, which includes the amino acid sequence shown in any one of SEQ ID NOs:2-5, and HC-CDR3, which includes SEQ ID NOs:6-11 The amino acid sequence shown in any one of; and b) a light chain variable domain, the light chain variable domain comprising: LC-CDR1, which includes the amino acid sequence shown in any one of SEQ ID NOs: 12-16 , LC-CDR2, which includes the amino acid sequence shown in any one of SEQ ID NOs: 17-19, and LC-CDR3, which includes the amino acid sequence shown in any one of SEQ ID NOs: 20-24.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO:1, HC-CDR2, which includes the amino acid sequence shown in any one of SEQ ID NOs:2-5, and HC-CDR3, which includes SEQ ID NOs:6-11 The amino acid sequence shown in any one of; and b) a light chain variable domain, the light chain variable domain comprising: LC-CDR1, which includes the amino acid sequence shown in any one of SEQ ID NOs: 12-16 , LC-CDR2, which includes the amino acid sequence shown in any one of SEQ ID NOs: 17-19, and LC-CDR3, which includes the amino acid sequence shown in any one of SEQ ID NOs: 20-24.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 2, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 6; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO:20.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO:20.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 8; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 13, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty one.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 5, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 10; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty three.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 15, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 11; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 16, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 19, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty four.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 2, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 6; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO:20.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO:20.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 8; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 13, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty one.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 5, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 10; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty three.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 15, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 11; and b) a light chain variable domain,
  • the light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 16, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 19, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty four.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region wherein the anti-C5a antibody comprises: a heavy chain variable domain ( VH ), the VH comprising SEQ ID NOs: 25-34 Any amino acid sequence shown or a variant thereof, which variant has at least about 80% (such as at least 80%, 85%, 90%, 95%) with the amino acid sequence shown in any one of SEQ ID NOs: 25-34 %, 96%, 97%, 98% or 99% ) sequence identity; and a light chain variable domain ( VL ) comprising the amino acid sequence shown in any one of SEQ ID NOs: 35-44 Or a variant thereof that shares at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%) with the amino acid sequence shown in any one of SEQ ID NOs: 35-44 , 98% or 99%) sequence identity.
  • VH heavy chain variable domain
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG2 constant region wherein the anti-C5a antibody comprises: a heavy chain variable domain ( VH ), the VH comprising SEQ ID NOs: 25-34 Any amino acid sequence shown or a variant thereof, which variant has at least about 80% (such as at least 80%, 85%, 90%, 95%) with the amino acid sequence shown in any one of SEQ ID NOs: 25-34 %, 96%, 97%, 98% or 99%) sequence identity; and a light chain variable domain ( VL ) comprising the amino acid sequence shown in any one of SEQ ID NOs: 35-44 Or a variant thereof that shares at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%) with the amino acid sequence shown in any one of SEQ ID NOs: 35-44 , 98% or 99%) sequence identity.
  • VH heavy chain variable domain
  • the IgG2 is human IgG2.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG3 constant region wherein the anti-C5a antibody comprises: a heavy chain variable domain ( VH ), the VH comprising SEQ ID NOs: 25-34 Any amino acid sequence shown or a variant thereof, which variant has at least about 80% (such as at least 80%, 85%, 90%, 95%) with the amino acid sequence shown in any one of SEQ ID NOs: 25-34 %, 96%, 97%, 98% or 99% ) sequence identity; and a light chain variable domain ( VL ) comprising the amino acid sequence shown in any one of SEQ ID NOs: 35-44 Or a variant thereof that shares at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%) with the amino acid sequence shown in any one of SEQ ID NOs: 35-44 , 98% or 99%) sequence identity.
  • VH heavy chain variable domain
  • the IgG3 is human IgG3.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region wherein the anti-C5a antibody comprises: a heavy chain variable domain ( VH ), the VH comprising SEQ ID NOs: 25-34 Any amino acid sequence shown or a variant thereof, which variant has at least about 80% (such as at least 80%, 85%, 90%, 95%) with the amino acid sequence shown in any one of SEQ ID NOs: 25-34 %, 96%, 97%, 98% or 99% ) sequence identity; and a light chain variable domain ( VL ) comprising the amino acid sequence shown in any one of SEQ ID NOs: 35-44 Or a variant thereof that shares at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%) with the amino acid sequence shown in any one of SEQ ID NOs: 35-44 , 98% or 99%) sequence identity.
  • VH heavy chain variable domain
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgGl constant region comprising an IgGl constant region
  • the anti-C5a antibody comprises: a heavy chain variable domain ( VH ) comprising SEQ ID NOs: 25- The amino acid sequence shown in any one of SEQ ID NOs: 35-44, and the light chain variable domain (V L ), said (V L ) comprising the amino acid sequence shown in any one of SEQ ID NOs: 35-44.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region wherein the anti-C5a antibody comprises: a heavy chain variable domain ( VH ), the VH comprising SEQ ID NOs: 25-34 The amino acid sequence shown in any one, and the light chain variable domain ( VL ), the VL comprising the amino acid sequence shown in any one of SEQ ID NOs: 35-44.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 25 or a variant thereof, the variant having the same amino acid sequence SEQ ID NO:25 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:35 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:35 Identity.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 26 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:26 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:36 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:36 Identity.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 27 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:27 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:37 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:37 Identity.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 28 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:28 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:38 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:38 Identity.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 29 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:29 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:39 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:39 Identity.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 30 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:30 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:40 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:40 Identity.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 31 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:31 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:41 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:41 Identity.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region comprising: V H comprising the amino acid sequence SEQ ID NO: 32 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:32 having at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:42 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:42 Identity.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 33 or a variant thereof, the variant is identical to the amino acid sequence SEQ ID NO:33 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:43 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:43 Identity.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG1 constant region wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 34 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:34 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:44 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:44 Identity.
  • the IgG1 is human IgG1.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: V H comprising the amino acid sequence SEQ ID NO: 25 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:25 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:35 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:35 Identity.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: V H comprising the amino acid sequence SEQ ID NO: 26 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:26 having at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:36 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:36 Identity.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: V H comprising the amino acid sequence SEQ ID NO: 27 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:27 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:37 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:37 Identity.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: V H comprising the amino acid sequence SEQ ID NO: 28 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:28 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:38 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:38 Identity.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: V H comprising the amino acid sequence SEQ ID NO: 29 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:29 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:39 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:39 Identity.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: V H comprising the amino acid sequence SEQ ID NO: 30 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:30 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:40 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:40 Identity.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising an IgG4 constant region, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 31 or a variant thereof, the variant having the same amino acid sequence SEQ ID NO:31 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:41 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:41 Identity.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: V H comprising the amino acid sequence SEQ ID NO: 32 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:32 having at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:42 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:42 Identity.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: V H comprising the amino acid sequence SEQ ID NO: 33 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:33 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:43 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:43 Identity.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • a full-length anti-C5a antibody comprising an IgG4 constant region comprising: V H comprising the amino acid sequence SEQ ID NO: 34 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:34 has at least about 80% sequence identity; and V L comprising the amino acid sequence SEQ ID NO:44 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:44 Identity.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
  • Binding affinity is expressed as Kd, Koff, Kon or Ka.
  • Koff refers to the rate constant for the dissociation of an antibody from an antigen/antibody complex, as measured by a kinetic selection device.
  • Kon refers to the binding rate constant at which an antibody binds to an antigen to form an antigen/antibody complex.
  • the equilibrium dissociation constant Kd used in this article refers to the dissociation constant when a specific antibody-antigen interacts. It refers to the antigen concentration required when the antigen occupies half of all antibody binding sites in the antibody molecule solution and reaches equilibrium, which is equal to Koff /Kon. The determination of Kd assumes that all bound molecules are in solution.
  • the corresponding equilibrium dissociation rate constant is expressed as EC50, which is a good approximation of Kd.
  • the affinity binding constant Ka is the reciprocal of the dissociation constant Kd.
  • the dissociation constant (Kd) can be used as an indicator of the affinity of the reactive antibody moiety to the antigen.
  • Kd dissociation constant
  • a simple analysis can be performed by the Scatchard method using antibodies labeled with various markers and a Biacore instrument (manufactured by Amersham Biosciences) to analyze interactions between biomolecules by surface plasmon resonance according to the user manual or the included kit. .
  • the Kd value obtained using these methods is expressed in the unit M.
  • Antibodies that specifically bind a target may have, for example, ⁇ 10 -7 M, ⁇ 10 -8 M, ⁇ 10 -9 M, ⁇ 10 -10 M, ⁇ 10 -11 M , ⁇ 10 -12 M, or ⁇ 10 - Kd value of 13 M.
  • the binding specificity of an antibody can be determined experimentally by methods known in the art. These methods include, but are not limited to, Western blots, ELISA-, RIA-, ECL-, IRMA-, EIA-, BIAcore testing, and peptide scanning.
  • the anti-C5a antibody specifically binds to the C5a target with a Kd value of 10 -7 M to 10 -13 M (e.g., 10 -7 M to 10 -13 M, 10 -8 M to 10 -13 M, 10 -9 M to 10 -13 M or 10 -10 M to 10 -12 M).
  • the Kd value of the binding between the anti-C5a antibody and C5a is 10 -7 M to 10 -13 M, 1 ⁇ 10 -7 M to 5 ⁇ 10 -13 M, 10 -7 M to 10 -12 M, 10 -7 M to 10 -11 M, 10 -7 M to 10 -10 M, 10 -7 M to 10 -9 M, 10 -8 M to 10 -13 M , 1 ⁇ 10 -8 M to 5 ⁇ 10 -13 M, 10 -8 M to 10 -12 M, 10 -8 M to 10 -11 M, 10 -8 M to 10 -10 M, 10 -8 M to 10 -9 M, 5 ⁇ 10 -9 M to 1 ⁇ 10 -13 M, 5 ⁇ 10 -9 M to 1 ⁇ 10 -12 M, 5 ⁇ 10 -9 M to 1 ⁇ 10 -11 M, 5 ⁇ 10 -9 M to 1 ⁇ 10 -10 M, 10 -9 M to 10 -13 M, 10 -9 M to 10 -12 M, 10 -9 M to 10 -12 M, 10 -9 M to 10 -11 M, 10 -9 M to 10 -10 M
  • the Kd value for binding between the anti-C5a antibody and the non-target is higher than the Kd value for the anti-C5a antibody binding to the target, and in some embodiments cited herein, the Kd value for the anti-C5a antibody binding to the target (e.g., C5a)
  • the binding affinity is higher than the binding affinity of the anti-C5a antibody to the non-target.
  • non-target refers to an antigen other than C5a.
  • the Kd value of the anti-C5a antibody (for C5a) binding to the non-C5a target is at least 10-fold different, such as 10-100 times, 100-1000 times, 10 3 -10 4 times, 10 4 -10 5 times, 10 5 -10 6 times, 10 6 -10 7 times, 10 7 -10 8 times, 10 8 -10 9 times , 10 9 -10 10 times, 10 10 -10 11 times, 10 11 -10 12 times .
  • the anti-C5a antibody binds to a non-target with a Kd value of 10 -1 M to 10 -6 M (e.g., 10 -1 M to 10 -6 M, 10 -1 M to 10 -5 M, 10 -2 M to 10 -4 M).
  • the non-target refers to an antigen other than C5a.
  • the Kd value for binding between the anti-C5a antibody and the non-C5a target is 10 -1 M to 10 -6 M, 1 ⁇ 10 -1 M to 5 ⁇ 10 -6 M, 10 -1 M to 10 -5 M, 1 ⁇ 10 -1 M to 5 ⁇ 10 -5 M, 10 -1 M to 10 -4 M, 1 ⁇ 10 -1 M to 5 ⁇ 10 -4 M, 10 - 1 M to 10 -3 M, 1 ⁇ 10 -1 M to 5 ⁇ 10 -3 M, 10 -1 M to 10 -2 M, 10 -2 M to 10 -6 M, 1 ⁇ 10 -2 M to 5 ⁇ 10 -6 M, 10 -2 M to 10 -5 M, 1 ⁇ 10 -2 M to 5 ⁇ 10 -5 M, 10 -2 M to 10 -4 M, 1 ⁇ 10 -2 M to 5 ⁇ 10 -4 M, 10 -2 M to 10 -3 M, 10 -3 M to 10 -6 M, 1 ⁇ 10 -3 M to 5 ⁇ 10 -6 M, 10 -3 M to 10 -6 M, 1 ⁇ 10 -3 M to 10
  • an anti-C5a antibody when it is referred to that an anti-C5a antibody specifically recognizes a C5a target with high binding affinity and binds to a non-target with low binding affinity, the anti-C5a antibody binds to the C5a target with a Kd value of 10 -7 M to 10 -13 M (for example, 10 -7 M to 10 -13 M, 10 -8 M to 10 -13 M, 10 -9 M to 10 -13 M, 10 -10 M to 10 -12 M), and with The Kd value for non-target binding is 10 -1 M to 10 -6 M (eg, 10 -1 M to 10 -6 M, 10 -1 M to 10 -5 M, 10 -2 M to 10 -4 M).
  • the binding affinity of the anti-C5a antibody is compared to the binding affinity of a control anti-C5a antibody (eg, INab308).
  • the Kd value of the binding between the control anti-C5a antibody and C5a can be at least 2 times, such as 2 times, 3 times, 4 times the Kd value of the binding between the anti-C5a antibody and C5a described herein. , 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 10-100 times, 100-1000 times, 10 3 -10 4 times.
  • nucleic acid molecules encoding anti-C5a antibodies are also considered.
  • a nucleic acid (or set of) nucleic acids encoding a full-length anti-C5a antibody is provided, including any of the full-length anti-C5a antibodies described herein.
  • the nucleic acid (or set of nucleic acids) of the anti-C5a antibody described herein may also include a nucleic acid sequence encoding a polypeptide tag (eg, protein purification tag, His tag, HA tag).
  • isolated host cells comprising an anti-C5a antibody, an isolated nucleic acid encoding an anti-C5a antibody polypeptide component, or a vector comprising a nucleic acid encoding an anti-C5a antibody polypeptide component described herein.
  • variants include nucleotide sequences that hybridize under at least moderately stringent hybridization conditions to a nucleic acid sequence encoding an anti-C5a antibody of the present application.
  • the present application also provides a vector into which the nucleic acid sequence of the present application can be inserted.
  • a natural or synthetic nucleic acid encoding an anti-C5a antibody is inserted into a suitable expression vector such that the nucleic acid is operably linked to 5' and 3' regulatory elements, such as a promoter (e.g., lymphocyte-specific promoter) and 3' untranslated region (UTR), which can express anti-C5a antibodies (such as full-length anti-C5a antibodies).
  • the vector may be suitable for replication and integration in eukaryotic host cells.
  • Typical cloning and expression vectors contain transcriptional and translational terminators, initiation sequences, and promoters that regulate expression of the nucleic acid sequence of interest.
  • nucleic acids described herein can also be used for nucleic acid immunization and gene therapy using standard gene delivery protocols.
  • Nucleic acid delivery methods are known in the art. See, for example, U.S. Pat. Nos. 5,399,346, 5,580,859, 5,589,466, the entire contents of which are incorporated herein by reference.
  • the application also provides gene therapy vectors.
  • Nucleic acids can be cloned into many types of vectors.
  • nucleic acids can be cloned into vectors including, but not limited to, plasmids, phagemids, phage derivatives, animal viruses, and cosmids.
  • Vectors of particular interest include expression vectors, replication vectors, probe generation vectors and sequencing vectors.
  • expression vectors can be provided to cells in the form of viral vectors.
  • Viral vector technology is well known in the art and is described, for example, in Green and Sambrook (2013, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York), and other virology or molecular biology manuals.
  • Viruses that can be used as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpesviruses, and lentiviruses.
  • suitable vectors include an origin of replication functional in at least one organism, promoter sequences, convenient restriction enzyme sites, and one or more selectable markers (see, e.g., WO 01/96584; WO 01/29058; and U.S. Pat. No. 6,326,193).
  • retroviruses provide a convenient platform for gene delivery systems.
  • the selected genes can be inserted into vectors and packaged in retroviral particles using techniques known in the art.
  • the recombinant virus is then isolated and delivered to the subject's cells in vivo or in vitro.
  • Many retroviral systems are known in the art.
  • adenoviral vectors are used.
  • Many adenoviral vectors are known in the art.
  • lentiviral vectors are used.
  • Vectors derived from retroviruses, such as lentiviruses are suitable tools for long-term gene transfer because they enable long-term stable integration of the transgene and propagation in progeny cells.
  • Lentiviral vectors have an additional advantage over tumor-derived retroviruses such as murine leukemia virus in that they can transduce non-dividing cells such as hepatocytes. At the same time, it has the additional advantage of low immunogenicity.
  • promoter elements such as enhancers, regulate the frequency of transcription initiation. Typically they are located 30-110 bp upstream of the start site, although recently many promoters have been found to also contain functional elements downstream of the start site.
  • the spacing between promoter elements is usually flexible, so that promoter function is maintained when elements are interchanged or moved with each other. In the thymidine kinase (tk) promoter, activity does not begin to decrease until the spacing between promoter elements increases to 50 bp.
  • tk thymidine kinase
  • a suitable promoter is the immediate early cytomegalovirus (CMV) promoter sequence. This promoter sequence is a strong constitutive promoter sequence that can drive high-level expression of any polynucleotide sequence operably linked to it.
  • CMV immediate early cytomegalovirus
  • EF-1 ⁇ elongation factor 1 ⁇
  • constitutive promoters may also be used, including, but not limited to, simian virus 40 (SV40) early promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus long terminal repeat (HIV-LTR) promoter , MoMuLV promoter, avian leukemia virus promoter, Epstein-Barr virus immediate early promoter, Rous sarcoma virus promoter and human gene promoters, such as but not limited to actin promoter, myosin promoter, Hemoglobin promoter and creatine kinase promoter.
  • SV40 simian virus 40
  • MMTV mouse mammary tumor virus
  • HV-LTR human immunodeficiency virus long terminal repeat
  • MoMuLV promoter avian leukemia virus promoter
  • Epstein-Barr virus immediate early promoter Epstein-Barr virus immediate early promoter
  • Rous sarcoma virus promoter Rous sarcoma virus promoter
  • human gene promoters such as but not
  • an inducible promoter provides a molecular switch that turns on the expression of the polynucleotide sequence to which it is operably linked when such expression is desired, and turns off expression when it is not.
  • Inducible promoters include, but are not limited to, metallothionein promoters, glucocorticoid promoters, progesterone promoters, and tetracycline promoters.
  • expression of anti-C5a antibodies is inducible.
  • a nucleic acid sequence encoding an anti-C5a antibody is operably linked to an inducible promoter, including any of the inducible promoters described herein.
  • an inducible promoter provides a molecular switch that turns on the expression of a polynucleotide sequence operably linked to it when expression is desired, and turns off expression when expression is not needed.
  • exemplary inducible promoters suitable for use in eukaryotic cells include, but are not limited to, hormone regulatory elements (see, e.g., Mader, S. and White, J.H. (1993) Proc. Natl. Acad. Sci. USA 90:5603-5607) , synthetic ligand regulatory elements (see Spencer, D.M. et al (1993) Science 262:1019-1024) and ionizing radiation regulatory elements (see Manome, Y. et al.
  • inducible promoter system used to express anti-C5a antibodies is the Tet system.
  • inducible promoter system used to express anti-C5a antibodies is an E. coli lac repressor system.
  • Tet system An exemplary inducible promoter system used in this application is the Tet system. This system is based on the Tet system described by Gossen et al. (1993).
  • the target polynucleotide is controlled by a promoter containing one or more Tet operator (TetO) sites.
  • TetO Tet operator
  • TetR Tet repressor
  • the activated state for example, in the presence of inducers such as tetracycline (Tc), anhydrotetracycline, doxycycline (Dox) or their active analogs, the inducer causes TetR to be released from TetO, resulting in transcription. .
  • Doxycycline is a member of the tetracycline antibiotic family, its chemical name is 1-dimethylamino-2,4a,5,7-pentahydroxy-11-methyl-4,6-dioxy-1,4a ,11,11a,12,12a-hexahydrotetraene-3-carboxamide.
  • TetR is codon-optimized for expression in mammalian cells, such as mouse or human cells. Due to the degeneracy of the genetic code, most amino acids are encoded by more than one codon, resulting in a large number of variants in the sequence of a given nucleic acid without any change in the sequence of its encoded amino acid. However, many organisms have differences in codon usage, also known as "codon preference" (i.e., the preference for a given amino acid to use a specific codon). Codon preference is often associated with the presence of dominant tRNA species for specific codons, which in turn increases the efficiency of mRNA translation. Coding sequences derived from a specific species (e.g., prokaryotes) can thus be tailored through codon optimization to enhance their expression in different species (e.g., eukaryotes).
  • a specific species e.g., prokaryotes
  • Tet-Off transcription is inactive in the presence of Tc or Dox.
  • tTA tetracycline-regulated transcriptional activator
  • TRE tetracycline-responsive promoter element
  • rtTA is a fusion protein composed of the TetR repressor and the VP16 transcriptional activation domain.
  • a 4-amino acid change in the DNA-binding region of TetR changes the binding properties of rtTA, causing it to only recognize the tetO sequence on the target transgene TRE in the presence of Dox. Therefore, in the Tet-On system, rtTA can activate the transcription of TRE-regulated target genes only in the presence of Dox.
  • lac repressor system of E. coli (see Brown et al., Cell 49:603-612 (1987)).
  • the Lac repressor system functions by regulating the transcription of a target polynucleotide operably linked to a promoter containing the lac operator (lacO).
  • lacO lac operator
  • LacR Lac repressor
  • lacR binds to LacO, thereby preventing the transcription of the target polynucleotide.
  • Expression of the polynucleotide of interest is induced by a suitable inducer, for example, isopropyl- ⁇ -D thiogalactopyranoside (IPTG).
  • IPTG isopropyl- ⁇ -D thiogalactopyranoside
  • the expression vector to be introduced into the cell may also contain a selectable marker gene or a reporter gene or both to facilitate the identification and selection of expressing cells from a population of cells transfected or infected with the viral vector.
  • the selectable marker can be carried on separate DNA fragments and used in co-transfection experiments.
  • Either the selectable marker gene or the reporter gene can be flanked by appropriate regulatory sequences to enable expression in the host cell.
  • Useful selectable markers include, for example, antibiotic resistance genes such as neo and similar genes.
  • Reporter genes can be used to identify potentially transfected cells and evaluate the function of regulatory sequences.
  • a reporter gene is a gene that is not present in or expressed by the recipient organism or tissue and encodes a polypeptide whose expression is manifested by some readily detectable property, such as enzymatic activity. After the DNA is introduced into the recipient cells, the expression of the reporter gene is detected at the appropriate time.
  • Suitable reporter genes may include genes encoding luciferase, ⁇ -galactosidase, chloramphenicol acetyltransferase, secreted alkaline phosphatase, or green fluorescent protein (see, Ui-Tel et al., 2000 FEBS Letters 479 :79-82).
  • Suitable expression systems are well known and can be prepared by known techniques or commercially available.
  • the construct with the smallest 5' flanking region that shows the highest expression level of the reporter gene is considered the promoter.
  • Such promoter regions can be linked to reporter genes and used to assess the ability of certain substances to regulate promoter-driven transcription.
  • nucleic acids encoding any of the full-length anti-C5a antibodies described herein are provided.
  • the nucleic acid includes one or more nucleic acid sequences encoding full-length anti-C5a antibody heavy and light chains.
  • each of the one or more nucleic acid sequences is contained in a separate vector.
  • at least some of the nucleic acid sequences are included in the same vector.
  • all nucleic acid sequences are contained in the same vector.
  • Vectors may be selected, for example, from mammalian expression vectors and viral vectors (eg, vectors derived from retroviruses, adenoviruses, adeno-associated viruses, herpesviruses and lentiviruses).
  • mammalian expression vectors and viral vectors eg, vectors derived from retroviruses, adenoviruses, adeno-associated viruses, herpesviruses and lentiviruses.
  • the vector can be readily introduced into host cells, such as mammalian cells, bacteria, yeast or insect cells, by any method in the art.
  • expression vectors can be introduced into host cells by physical, chemical or biological methods.
  • polynucleotides are introduced into the host cell by calcium phosphate transfection.
  • Biological methods for introducing polynucleotides of interest into host cells include the use of DNA and RNA vectors.
  • Viral vectors especially retroviral vectors, have become the most widely used method for inserting genes into mammalian cells, such as human cells.
  • Other viral vectors can be derived from lentivirus, poxvirus, herpes simplex virus type 1, adenovirus, adeno-associated virus, etc. See, for example, U.S. Pat. Nos. 5,350,674 and 5,585,362.
  • colloidal dispersion systems such as polymer complexes, nanocapsules, microspheres, magnetic beads, and lipid-based systems including oil-in-water emulsions, micelles, and mixed gels pellets and liposomes.
  • colloidal dispersion systems such as polymer complexes, nanocapsules, microspheres, magnetic beads, and lipid-based systems including oil-in-water emulsions, micelles, and mixed gels pellets and liposomes.
  • liposomes eg, artificial membrane vesicles.
  • an exemplary delivery vehicle is liposomes.
  • lipid formulations to introduce nucleic acids into host cells (in vitro, ex vivo, or in vivo).
  • the nucleic acid can be associated with lipids.
  • the nucleic acid bound to the lipid can be packaged into the aqueous interior of the liposome, spread within the lipid bilayer of the liposome, and connected to the liposome through a linker molecule that binds to the liposome and the oligonucleotide.
  • Lipid, lipid/DNA or lipid/expression vector related compositions are not limited to any particular structure in solution. For example, they may exist in bilayer structures, in micelles or in "collapsed" structures. They can also simply disperse in solution, possibly forming aggregates that are not uniform in size or shape. Lipids are fatty substances that can be naturally occurring or synthetic lipids.
  • lipids include lipid droplets that occur naturally in the cytoplasm, as well as a class of compounds containing long-chain aliphatic hydrocarbons and their derivatives, such as fatty acids, alcohols, amines, aminoalcohols, and aldehydes.
  • experiments can be performed in order to confirm that the recombinant DNA sequence is present in the host cell.
  • Such experiments include, for example, "molecular biology” experiments well known to those skilled in the art. For example, Southern and Northern blotting, RT-PCR and PCR; "biochemical” experiments, such as detecting the presence or absence of a specific polypeptide, such as by immunological methods (ELISAs and Western blots) or by experiments described in this article All fall within the scope of this application.
  • the anti-C5a antibody is a monoclonal antibody or is derived from a monoclonal antibody. In some embodiments, the anti-C5a antibody comprises VH and VL from a monoclonal antibody, or a variant thereof. In some embodiments, the anti-C5a antibody further includes CH1 and CL regions from a monoclonal antibody, or variants thereof.
  • Monoclonal antibodies can be prepared using, for example, methods known in the art, including hybridoma cell methods, phage display methods, or using recombinant DNA methods. Additionally, exemplary phage display methods are described herein and in the Examples below.
  • an immune agent is usually used to immunize hamsters, mice or other suitable host animals to induce lymphocytes that produce or are capable of producing antibodies that specifically bind to the immune agent.
  • lymphocytes can be immunized in vitro.
  • Immunizing agents may include polypeptides or fusion proteins of the protein of interest.
  • PBLs peripheral blood lymphocytes
  • spleen cells or lymph node cells are used.
  • Lymphocytes are fused to immortalized cell lines using an appropriate fusion agent, such as polyethylene glycol, to form hybridoma cells.
  • Immortal cell lines are typically transformed mammalian cells, particularly myeloma cells of rodent, bovine, and human origin. Typically, rat or mouse myeloma cell lines are used.
  • Hybridoma cells can be cultured in a suitable medium, which preferably contains one or more substances that inhibit the growth or survival of unfused immortal cells. For example, if the parent cells lack the enzyme hypoxanthine-guanine phosphoribosyltransferase (HGPRT or HPRT), the hybridoma cell culture medium usually includes hypoxanthine, aminopterin, and thymidine (HAT medium), which can Prevents the growth of HGPRT-deficient cells.
  • HGPRT hypoxanthine-guanine phosphoribosyltransferase
  • the immortalized cell lines fuse efficiently, ensure high-level and stable expression of the antibody by the selected antibody-producing cells, and are sensitive to certain media, such as HAT media.
  • the immortal cell line is a mouse myeloma cell line, available from, for example, the Salk Cell Collection in San Diego, California, and the American Type Culture Collection in Manassas, Virginia. Also described are human myeloma and mouse-human hybrid myeloma cell lines for the preparation of human monoclonal antibodies.
  • the culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against the polypeptide.
  • the binding specificity of monoclonal antibodies produced by hybridoma cells can be determined by immunoprecipitation or in vitro binding experiments, such as radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). Such techniques or analytical methods are known in the art.
  • the binding affinity of a monoclonal antibody can be determined by Scatchard analysis as described, for example, in Munson and Pollard, Anal. Biochem., 107:220 (1980).
  • hybridoma cells After the desired hybridoma cells are identified, the clones of interest can be subcloned by limiting dilution and cultured by standard methods. Suitable media for this purpose include, for example, modified Eagle's medium (DMEM) and RPMI-1640 medium. Alternatively, hybridoma cells can be grown in mammals in the form of ascites fluid.
  • DMEM modified Eagle's medium
  • RPMI-1640 medium RPMI-1640
  • Monoclonal antibodies secreted by the subclones can be isolated or purified from the culture medium or ascitic fluid by conventional immunoglobulin purification methods, such as protein A-Sepharose, hydroxyapatite chromatography, gel electrophoresis, dialysis, or affinity Chromatography.
  • immunoglobulin purification methods such as protein A-Sepharose, hydroxyapatite chromatography, gel electrophoresis, dialysis, or affinity Chromatography.
  • the anti-C5a antibody comprises sequences selected from clones of an antibody library (eg, a phage library displaying scFv or Fab fragments).
  • the clones can be identified by screening a combinatorial library of antibody fragments with the desired activity.
  • various methods are known in the art for generating phage display libraries and screening these libraries for antibodies with desired binding properties. These methods are reviewed, for example, in Hoogenboom et al., Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, N.J., 2001), and in, e.g., McCafferty et al.
  • phage display methods all components of the V H and V L genes are cloned separately through polymerase chain reaction (PCR), randomly recombined in a phage library, and then screened for phages that can bind the antigen, such as Winter et al. ., Ann. Rev. Immunol., 12:433-455 (1994). Phages typically display antibody fragments as scFv fragments or as Fab fragments. Immunogenically derived library phages provide high-affinity antibodies against immunogens without the need to construct hybridoma cells.
  • PCR polymerase chain reaction
  • natural libraries can be cloned to provide a single source of antibodies against multiple non-self and self-antigens without the need for any immunization, as in Griffiths et al., EMBOJ, 12:725-734 (1993) described in.
  • natural libraries can also be prepared by cloning non-rearranged V-gene fragments from stem cells and using PCR primers containing random sequences to encode the CDR3 hypervariable region and completing the rearrangement in vitro, such as Hoogenboom and Winter, J. Mol. Biol., 227:381-388 (1992).
  • Patent publications describing human antibody phage libraries include, for example, US Pat. No. 5,750,373 and US Patent Publication Nos. 2005/0079574, 2005/0119455, 2005/0266000, 2007/0117126, 2007/0160598, 2007/0237764, 2007/0 292936 and 2009/0002360.
  • the anti-C5a antibody is prepared by phage display screening library for anti-C5a antibody portions that can specifically bind to target C5a.
  • the library may be a human scFv phage display library with at least 1 ⁇ 10 9 (e.g., at least 1 ⁇ 10 9 , 2.5 ⁇ 10 9 , 5 ⁇ 10 9 , 7.5 ⁇ 10 9 , 1 ⁇ 10 10 , 2.5 ⁇ 10 10 , 5 ⁇ 10 10 , 7.5 ⁇ 10 10 or 1 ⁇ 10 11 ) diversity of unique human antibody fragments.
  • the library is a human natural library, constructed from DNA extracted from PMBCs and spleens of healthy subjects, containing all human heavy and light chain subfamilies.
  • the library is a human natural library constructed from DNA extracted from PMBCs isolated from patients with various diseases, such as patients with autoimmune diseases, cancer patients, and patients with infectious diseases.
  • the library is a semi-synthetic human library in which the heavy chain CDR3s are completely random and all amino acids (except cysteine) are present with equal probability at any given position. (See, eg, Hoet, RM et al., Nat. Biotechnol. 23(3):344-348, 2005).
  • the heavy chain CDR3 length of the semi-synthetic human library ranges from 5 to 24 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 , 19, 20, 21, 22, 23 or 24) between amino acids.
  • the library is a fully synthetic phage display library.
  • the library is a non-human phage display library.
  • Phage clones with high affinity for target C5a can be screened by iterative binding of phage to target C5a bound to a solid support (e.g., beads for solution panning or mammalian cells for cell panning) ), followed by removal of unbound phage and elution of specifically bound phage. Subsequently, bound phage clones are eluted and used to infect suitable host cells, such as E. coli XL1-Blue, for expression and purification. Phage clones that specifically bind C5a can be enriched by multiple rounds of panning (eg, 2, 3, 4, 5, 6, or more rounds), such as solution panning, cell panning, or a combination of both. Specific binding of the enriched phage clones to the target C5a can be detected by any method known in the art, including, for example, ELISA and FACS.
  • Another way to screen antibody libraries is to display proteins on the surface of yeast cells.
  • Wittrup et al. (US Patents 6,699,658 and 6,696,251) developed a method for displaying libraries in yeast cells.
  • one component includes the yeast lectin protein (Aga1), which is anchored to the yeast cell wall, and the other component includes the second subunit of the lectin protein Aga2, which can pass through disulfide bonds. Binds to the Aga1 protein and is displayed on the surface of yeast cells.
  • Aga1 protein is expressed by integrating the Aga1 gene into the yeast chromosome.
  • a single-chain variable fragment (scFv) library is fused to the Aga2 gene in a yeast display plasmid and, after transformation, is retained in yeast due to the presence of an additional nutritional marker. Both Aga1 and Aga2 proteins are expressed under the control of galactose-inducible promoters.
  • the human antibody V gene library (V H and V K fragments) was obtained by PCR using a set of degenerate primers (Sblattero, D. and Bradbury, A. Immunotechnology 3, 271-278 1998).
  • PCR templates were derived from commercially available RNA or cDNA, including PBMC, spleen, lymph nodes, bone marrow, and tonsils.
  • Independent V H and V K PCR libraries were combined and assembled into scFv formats by overlap extension PCR (Sheets, MD et al, Proc. Natl. Acad. Sci. USA 95, 6157–61621998).
  • yeast scFv display library To construct a yeast scFv display library, the resulting scFv PCR product was cloned into a yeast display plasmid in yeast via homologous recombination. (Chao, G, et al, Nat Protoc. 2006; 1(2):755-68. Miller KD, et al. Current Protocols in Cytometry 4.7.1-4.7.30, 2008).
  • Anti-C5a antibodies can be screened using a mammalian cell display system, in which the antibody moiety is displayed on the cell surface and antibodies specifically targeting C5a are isolated by antigen-directed screening methods (as described in U.S. patent No. 7,732,195B2).
  • a Chinese hamster ovary (CHO) cell library displaying a large number of human IgG antibody genes can be created and used to discover clones expressing high-affinity antibody genes.
  • An alternative display system has been developed that allows the same protein to be simultaneously displayed and secreted on the cell surface through alternative splicing, in which the phenotype of the displayed protein remains correlated with the genotype, allowing for simultaneous biophysical and cell function-based analyses. Characterize the secreted soluble antibodies.
  • This method overcomes many of the limitations of previous mammalian cell displays and enables direct screening and maturation of antibodies in the form of full-length, glycosylated IgGs (Peter M. Bowers, et al, Methods 2014, 65: 44-56).
  • Transient expression systems are suitable for a single round of antigen selection prior to antibody gene recovery and are therefore most useful for selecting antibodies from smaller libraries.
  • Stable exosome vectors offer an attractive option. Exosome vectors can be efficiently transfected and stably maintained at low copy numbers, allowing for multiple rounds of panning and elucidation of more complex antibody libraries.
  • the IgG library was constructed based on the ligation of germline sequence V gene fragments isolated from a pool of human donors and rearranged (D)J regions. RNA collected from 2000 human blood samples was reverse transcribed into cDNA, VH and VK fragments were amplified using VH and VK specific primers, and purified by gel extraction. The VH and VK fragments were subcloned into display vectors containing IgG1 or K constant regions respectively, and then electroporated or transduced 293T into cells to prepare IgG libraries.
  • V H and V K are ligated to generate scFv, which is then subcloned into a display vector and electroporated or transduced into 293T cells.
  • IgG libraries are constructed based on germline sequence V gene fragments and rearranged (D)J regions isolated from a group of donors, which can be mice, rats, rabbits or monkeys.
  • Monoclonal antibodies can also be prepared by recombinant DNA methods, such as those described in U.S. Patent No. 4,816,567.
  • DNA encoding the monoclonal antibodies described herein can be readily isolated and sequenced by conventional methods (eg, by oligonucleotide probes that specifically bind to genes encoding the light and heavy chains of murine antibodies).
  • Hybridoma cells as described above or the C5a-specific phage clones of the present application can be used as a source of such DNA.
  • the DNA can be placed into an expression vector, which can then be transfected into host cells, such as simian COS cells, Chinese hamster ovary cancer (CHO) cells, or myeloma cells that do not produce immunoglobulins, to obtain the expression in the recombinant host.
  • host cells such as simian COS cells, Chinese hamster ovary cancer (CHO) cells, or myeloma cells that do not produce immunoglobulins, to obtain the expression in the recombinant host.
  • Monoclonal antibodies synthesized in cells may also be modified, for example by substituting coding sequences for human heavy and light chain constant regions and/or substituting framework regions for homologous non-human sequences (U.S. Patent No.
  • non-immunoglobulin polypeptide is covalently linked to the coding sequence for an immunoglobulin.
  • This non-immunoglobulin polypeptide can replace the constant region of the antibody in this application, or can replace an antigen-binding site in the variable domain of the antibody in this application, forming a chimeric bivalent antibody.
  • the antibody may be a monovalent antibody.
  • Methods of preparing monovalent antibodies are known in the art. For example, one involves a recombinant expression method involving an immunoglobulin light chain and a modified heavy chain.
  • the heavy chain is usually truncated at any position in the Fc region to prevent the heavy chains from cross-linking with each other.
  • the relevant cysteine residues are substituted with other amino acid residues or deleted to prevent cross-linking.
  • In vitro methods are also suitable for preparing monovalent antibodies. Digestion of antibodies to produce antibody fragments, particularly Fab fragments, can be accomplished using any method known in the art.
  • Antibody variable domains with the desired binding specificity can be fused to immunoglobulin constant regions. Fusions to the immunoglobulin heavy chain constant region, including at least part of the hinge, CH2 and CH3 regions, are preferred. In some embodiments, the first heavy chain constant region (CH1), which contains the necessary sites for light chain binding, is present in at least one fusion. DNA encoding the immunoglobulin heavy chain fusion, and if desired, DNA encoding the immunoglobulin light chain, is inserted into a separate expression vector and co-transfected into a suitable host organism.
  • the anti-C5a antibody (eg, full-length anti-C5a antibody) can be a fully human antibody or a humanized antibody.
  • Humanized forms of non-human (e.g., mouse) antibody portions are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (e.g., Fv, Fab, Fab', F(ab') 2 , scFv, or other fragments of antibodies Antigen binder sequences), which generally include minimal sequences derived from non-human immunoglobulins.
  • Humanized antibodies include human immunoglobulins, immunoglobulin chains or fragments thereof (recipient antibodies) in which the residues of the acceptor CDRs are replaced by non-human (donor antibody) CDRs with the desired specificity, affinity and properties. Residue substitutions, such as mouse, rat or rabbit CDRs. In some embodiments, human immunoglobulin Fv framework residues are replaced with corresponding non-human residues. Humanized antibodies may also contain amino acid residues that neither belong to the recipient antibody nor are in the introduced CDR or framework sequence.
  • a humanized antibody contains at least one, and usually two variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the framework regions are common to human immunoglobulins. sequence.
  • humanized antibodies typically contain one or more amino acid residues introduced from a non-human source. Those non-human amino acid residues are often referred to as "implanted” residues, usually from the "imported” variable domain. According to some embodiments, humanization can be performed essentially as described by Winter and colleagues (Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 ( 1988); Verhoeyen et al., Science, 239:1534-1536 (1988)), by replacing the corresponding sequences of human antibodies with rodent CDRs or CDR sequences. Thus, this "humanized” antibody portion (U.S. Patent No.
  • a humanized antibody portion is a typical human antibody portion in which some CDR residues and possibly some framework region residues are replaced by residues from similar positions in rodent antibodies.
  • Fully human antibodies are an alternative to humanization. For example, it is now possible to generate transgenic animals (eg, mice) that are capable of producing a complete library of fully human antibodies upon immunization without producing endogenous immunoglobulins. For example, it has been reported that homozygous deletion of the antibody heavy chain junction region (JH) gene in chimeric and germline mutant mice completely inhibits endogenous antibody production.
  • JH antibody heavy chain junction region
  • Transferring human germline immunoglobulin gene arrays into such germline mutant mice can produce human antibodies in response to antigen stimulation, see, e.g., akobovits et al., PNAS USA, 90:2551 (1993); Jakobovits et al ., Nature, 362:255-258 (1993); Bruggemann et al., Year in Immunol., 7:33 (1993); U.S. Patent Nos. 5,545,806, 5,569,825, 5,591,669, 5,545,807; and WO 97/17852.
  • Fully human antibodies can be prepared by introducing human immunoglobulin loci into transgenic animals (eg, mice in which the endogenous immunoglobulin genes have been partially or completely silenced). After antigen stimulation, the production of fully human antibodies can be found to be very similar to their production in humans in all aspects, including gene rearrangement, assembly, and antibody libraries. This method is used in, for example, U.S. Patent Nos.
  • Fully human antibodies can also be produced by activating B cells in vitro (see U.S. Patents 5,567,610 and 5,229,275) or by using various techniques known in the art, including phage display libraries. Hoogenboom and Winter, J. Mol. Biol., 227: 381 (1991); Marks et al., J. Mol. Biol., 222: 581 (1991).
  • the technology of Cole et al. and Boerner et al. It can also be used to prepare fully human monoclonal antibodies. See Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985) and Boerner et al., J. Immunol., 147(1):86-95 (1991).
  • the amino acid sequences of anti-C5a antibody variants are also contemplated.
  • the amino acid sequences of antibody variants can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions and/or insertions and/or substitutions of residues in the antibody amino acid sequence.
  • the final construct can be accomplished by any combination of deletions, insertions, and substitutions of amino acid residues to give it the desired characteristics. For example, antigen binding.
  • anti-C5a antibody variants with one or more amino acid substitutions are provided.
  • Target sites for substitution mutations include hypervariable regions (HVRs) and framework regions (FRs).
  • Amino acid substitutions can be introduced into the antibody of interest and the product screened for the desired activity, for example, improved biological activity, maintained/improved antigen binding capacity, reduced immunogenicity, or improved ADCC or CDC.
  • Amino acids are divided into different categories based on the nature of their side chains:
  • Acidic amino acids aspartic acid Asp, glutamic acid Glu;
  • Aromatic amino acids tryptophan Trp, tyrosine Tyr, phenylalanine Phe.
  • Substitutions of non-conservative amino acids involve substitution of one category for another category.
  • An exemplary substitution variant is an affinity matured antibody, which can be conveniently produced using, for example, phage display-based affinity maturation techniques. Briefly, one or more CDR residues are mutated, variant antibody moieties are displayed on phage, and those screened for specific biological activity (e.g., biological activity based on a reactive oxygen species (ROS) release assay or binding affinity) Variants. Alterations (eg, substitutions) can be made in regions of HVRs to obtain improved biological activity or antibody affinity based on reactive oxygen species (ROS) release assays.
  • ROS reactive oxygen species
  • Alterations can occur in "hot spots" of HVR, i.e., codon-encoded residues that are highly mutated during somatic cell maturation (see, e.g., Chowdhury, Methods Mol. Biol. 207:179-196 (2008)), and/or detect the binding affinity of the resulting variants VH and VL at specific decisive residues (SDRs).
  • SDRs decisive residues
  • variable genes selected for affinity maturation are introduced into the variable genes selected for affinity maturation by any of a variety of methods, such as error-prone PCR, strand shuffling, or oligonucleotide-directed mutagenesis. .
  • Secondary libraries are then created. The library is screened to identify antibody variants with the desired affinity.
  • Another way to introduce diversity includes the HVR-mediated manner, in which several HVR residues (e.g., 4-6 residues at a time) are randomized. HVR residues involved in antigen binding are specifically recognized, for example, using alanine scanning mutagenesis or modeling. Usually the CDR-H3 and CDR-L3 regions are particularly focused targets.
  • substitutions, insertions, or deletions may occur within one or more HVRs, so long as such changes do not substantially reduce the ability of the antibody to bind the antigen.
  • conservative changes eg, conservative substitutions provided herein
  • HVRs that do not substantially reduce binding affinity. These changes may occur outside the HVR "hot zone" or SDRs area.
  • each HVR is either unchanged or contains no more than 1, 2, or 3 amino acid substitutions.
  • a useful method for identifying amino acid residues or regions of an antibody that can be targeted mutated is called "alanine scanning mutagenesis" as described in Cunningham and Wells (1989) Science, 244:1081-1085 .
  • target residues e.g., charged residues such as arginine, aspartic acid, histidine, lysine, and glutamic acid
  • neutral or negatively charged amino acids e.g., , alanine or glutamic acid
  • Further substitutions can be introduced at amino acid positions to demonstrate functional sensitivity of the position to the initial substitution.
  • the contact sites between the antibody and the antigen are identified through the crystal structure of the antigen-antibody complex. These contact site residues and adjacent residues can be targeted or eliminated as substitution candidates. Screen variants to determine if they have the desired properties.
  • Insertions of amino acid sequences including fusions at the amino and/or carboxyl terminus, ranging in length from 1 residue to polypeptides containing 100 or more residues, and including insertion of 1 or more amino acid residues within the sequence base.
  • terminal insertions include antibodies with a methionyl residue at the N-terminus.
  • Other insertional variants of antibody molecules include fusion of an enzyme (eg, ADEPT) or peptides that increase the serum half-life of the antibody to the N- or C-terminus of the antibody molecule.
  • one or more amino acid modifications are introduced into the Fc region of an antibody described herein (eg, a full-length anti-C5a antibody or an anti-C5a antibody fusion protein), thereby generating Fc region variants.
  • Fc region variants have enhanced ADCC potency, typically associated with binding to Fc receptors (FcRs).
  • Fc region variants have reduced ADCC efficacy.
  • changes or mutations in the Fc sequence affecting its potency For example, WO 00/42072 and Shields et al. J Biol. Chem. 9(2):6591-6604 (2001) describe enhanced binding to FcRs or Attenuated antibody variants. The contents of these publications are incorporated herein by reference.
  • ADCC Antibody-dependent cell-mediated cytotoxicity
  • NK cells activated by antibodies.
  • NK cells express the Fc receptor CD16. This receptor recognizes and binds to the Fc portion of the antibody molecule bound to the surface of the target cell.
  • the most common Fc receptors on the surface of NK cells are CD16 or Fc ⁇ RIII.
  • Binding of Fc receptors to the Fc region of antibodies results in activation of NK cells, release of cell lytic granules, and subsequent target cell apoptosis.
  • the killing effect of ADCC on tumor cells can be measured through specific experiments of NK-92 cells transfected with high-affinity FcR. The results were compared with wild-type NK-92, which does not express FcR.
  • the application also provides anti-C5a antibody variants (e.g., full-length anti-C5a antibody variants) that comprise an Fc region with some, but not all, of the effector functions such that they have an extended half-life in vivo, yet specifically
  • the effector function (such as CDC or ADCC) is non-essential or deleterious, making this anti-C5a antibody an ideal candidate for this application. Reduction/elimination of CDC and/or ADCC activity is confirmed by performing cytotoxicity assays in vitro and/or in vivo.
  • Fc receptor (FcR) binding assays are used to confirm that the antibody lacks Fc ⁇ R binding ability (and therefore may lack ADCC activity) but still retains FcRn binding ability.
  • NK cells only express Fc ⁇ RIII
  • monocytes express Fc ⁇ RI, Fc ⁇ RII, and Fc ⁇ RIII.
  • Table 3 The expression of FcR on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet Annu. Rev. Immunol. 9:457-492 (1991).
  • Non-limiting examples of in vitro assessment of ADCC activity of target molecules are described in US Pat. No. 5,500,362 (see, e.g., Hellstrom, I.
  • CYTOTOX 96 TM Nonradioactive Cell Toxicity assays (Promega, Madison, Wis.). Effector cells used in these assays include peripheral blood mononuclear cells (PBMC) and natural killer cells (NK).
  • PBMC peripheral blood mononuclear cells
  • NK natural killer cells
  • the ADCC activity of the target molecule is performed in vivo Detection, for example, in animal models, as described in Clynes et al. Proc. Nat'l Acad. Sci. USA 95:652-656 (1998).
  • a C1q binding assay can also be performed to confirm that the antibody cannot bind to C1q , thereby lacking CDC activity.
  • a CDC assay can be performed (see, for example, Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996); Cragg, MS et al., Blood 101:1045-1052 (2003); and Cragg, MS and MJ Glennie, Blood 103:2738-2743 (2004)). Determined using methods known in the art FcRn binding and clearance/half-life in vivo (see, eg, Petkova, SB et al., Int'l. Immunol. 18(12):1759-1769 (2006)).
  • Antibodies with reduced effector function include substitution of one or more residues at residues 238, 265, 269, 270, 297, 327, and 329 of the Fc region (U.S. Pat. No. 6,737,056). These Fc variants include Fc variants with substitutions of two or more residues at positions 265, 269, 270, 297 and 327, including the Fc variant known as "DANA" which has substitutions at residues 265 and 297. The base is substituted with alanine (U.S. Pat. No. 7,332,581).
  • an anti-C5a antibody eg, a full-length anti-C5a antibody
  • an Fc region variant with one or more amino acid substitutions capable of enhancing ADCC effects.
  • the Fc region variant contains one or more amino substitutions capable of enhancing the ADCC effect at positions 298, 333 and/or 334 (EU residue numbering) of the Fc region.
  • the anti-C5a antibody (eg, full-length anti-C5a antibody) variant includes amino acid substitutions at positions S298A, E333A, and K334A in the Fc region.
  • changes in the Fc region result in changes (i.e., enhancement or attenuation) of Clq binding and/or complement-dependent cytotoxicity (CDC), see U.S. Pat. No. 6,194,551, WO 99/51642, and Idusogie et al. al., J. Immunol. 164:4178-4184 (2000).
  • changes in the Fc region result in changes (i.e., enhancement or attenuation) of Clq binding and/or complement-dependent cytotoxicity (CDC), see U.S. Pat. No. 6,194,551, WO 99/51642, and Idusogie et al. al., J. Immunol. 164:4178-4184 (2000).
  • an anti-C5a antibody e.g., a full-length anti-C5a antibody
  • an Fc region variant with one or more amino acid substitutions capable of extending half-life or enhancing interaction with an Fc receptor (FcRn ) combination is provided that includes an Fc region variant with one or more amino acid substitutions capable of extending half-life or enhancing interaction with an Fc receptor (FcRn ) combination.
  • FcRn Fc receptor
  • Fc variants contain 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or One or more substitutions in residue 434, for example, substitution of residue 434 in the Fc region (U.S. Pat. No. 7,371,826).
  • anti-C5a antibodies eg, full-length anti-C5a antibodies
  • Fc variants described herein or combinations thereof.
  • anti-C5a antibodies provided herein are altered to increase or decrease the extent of glycosylation of the anti-NGF antibody. Adding or deleting glycosylation sites on the anti-C5a antibody can be conveniently achieved by changing the amino acid sequence of the anti-NGF antibody or its polypeptide portion to add or remove one or more glycosylation sites.
  • the anti-C5a antibody contains an Fc region that can change the sugar attached to it.
  • Natural antibodies produced by mammalian cells typically contain branched biantennary oligosaccharides that are often linked to the Fc region CH2 domain Asn297 via an N-link, see e.g. Wright et al., TIBTECH 15:26-32 (1997) .
  • the oligosaccharides can include a variety of sugars, such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as trehalose linked to GlcNAc in the "stem" portion of the biantennary oligosaccharide structure.
  • the anti-C5a antibodies of the present application can be subjected to oligosaccharide modifications, thereby producing anti-C5a antibody variants with certain improved properties.
  • N-glycans linked to the CH2 domain of the Fc region are heterogeneous.
  • Antibodies or Fc fusion proteins produced in CHO cells are fucosylated by fucosyltransferase activity, see Shoji-Hosaka et al., J. Biochem. 2006, 140:777-83.
  • a small proportion of naturally occurring afucosylated IgGs can be detected in human serum.
  • N-glycosylation of the Fc region is important for its binding to Fc ⁇ R; afucosylated N-glycan enhances the binding ability of Fc to Fc ⁇ RIIIa.
  • the enhanced binding ability to FcRIIIa results in enhanced ADCC effect, which is advantageous in certain antibody therapeutic applications that require cytotoxicity.
  • enhanced effector function may be detrimental when Fc-mediated cytotoxicity is not required.
  • the Fc fragment or CH2 domain is non-glycosylated.
  • glycosylation is prevented by mutating the N-glycosylation site in the CH2 domain.
  • anti-C5a antibody eg, full-length anti-C5a antibody
  • variants comprise an Fc region in which the carbohydrate structure linked to the Fc region has reduced fucose or lacks fucose, which may Will enhance ADCC function.
  • anti-C5a antibodies that have reduced fucose relative to the same anti-C5a antibody produced by wild-type CHO cells. That is, they are characterized by having smaller amounts of fucose than antibodies produced by native CHO cells (e.g., CHO cells that produce the native glycosylated form, CHO cells that contain the native FUT8 gene).
  • the N-linked glycan of the anti-C5a antibody has less than 50%, 40%, 30%, 20%, 10%, or 5% fucose.
  • the fucose content of the anti-C5a antibody may be 1%-80%, 1%-65%, 5%-65%, or 20%-40%.
  • the N-linked glycans of the anti-C5a antibody do not comprise fucose, i.e., wherein the anti-C5a antibody contains no fucose at all, or is free of fucose or afucosylated.
  • the fucose content is calculated by calculating the average fucose content within the sugar chain attached to Asn297 relative to the total of all sugar structures attached to Asn297 (such as complex, hybrid or mannose structures) measured by MALDI-TOF mass spectrometry. determined by quantity, as described in WO 2008/077546.
  • Asn297 refers to the asparagine residue located at position 297 in the Fc region (EU Fc region residue numbering system). However, due to minor sequence changes in the antibody, Asn297 can also be located ⁇ 3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300. These fucosylation variants may have enhanced ADCC functions. See, for example, US Patent Publication Nos.
  • Cell lines capable of producing afucosylated antibodies include Lec13 CHO cells lacking protein fucosylation function (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); US PatAppl No US 2003 /0157108 A1, Presta, L; and WO 2004/056312 A1, Adams et al., especially Example 11), and gene knockout cell lines, such as ⁇ -1,6-fucosyltransferase gene, FUT8 Gene knockout CHO cells (see Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94 (4): 680-688 (2006); and WO2003/085107).
  • Anti-C5a antibody (eg, full-length anti-C5a antibody) variants further provide bisecting oligosaccharides, eg, wherein the biantennary oligosaccharide linked to the Fc region of the anti-C5a antibody is bisected by GlcNAc.
  • Such anti-C5a antibody (eg, full-length anti-C5a antibody) variants may have reduced fucosylation and/or enhanced ADCC function. Examples of such antibody variants are in WO 2003/011878 (Jean-Mairet et al.); U.S. Pat. No. 6,602,684 (Umana et al.); US 2005/0123546 (Umana et al.), and Ferrara et al.
  • anti-C5a antibody eg, full-length anti-C5a antibody
  • anti-C5a antibody variants that have at least one galactose residue in the oligosaccharide linked to the Fc region.
  • Such anti-C5a antibody variants may have enhanced CDC function.
  • Such variants are described, for example, in WO 1997/30087 (Patel et al.); WO 1998/58964 (Raju, S.); and WO 1999/22764 (Raju, S.).
  • the anti-C5a antibody (eg, full-length anti-C5a antibody) variant comprises an Fc region capable of binding Fc ⁇ RIII.
  • the anti-C5a antibody (e.g., full-length anti-C5a antibody) variant comprising an Fc region has ADCC activity in the presence of human effector cells (e.g., T cells), or is identical to an anti-C5a antibody having a human wild-type IgG1 Fc region. Compared with other identical anti-C5a antibodies (eg, full-length anti-C5a antibodies), they have enhanced ADCC activity in the presence of human effector cells.
  • cysteine-engineered anti-C5a antibody eg, a full-length anti-C5a antibody
  • the substitution residue occurs at an accessible site of the anti-C5a antibody.
  • reactive thiol groups are located in accessible sites of the anti-C5a antibody and can be used to couple the anti-C5a antibody to other moieties, such as a drug moiety or a linker-drug moiety, to prepare anti-C5a immunoconjugates as further described herein.
  • Cysteine-engineered anti-C5a antibodies eg, full-length anti-C5a antibodies
  • anti-C5a antibodies eg, full-length anti-C5a antibodies
  • anti-C5a antibodies can be further modified to include other non-protein moieties known in the art and readily available.
  • Suitable moieties for derivatizing anti-C5a antibodies include, but are not limited to, water-soluble polymers.
  • Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol/propylene glycol copolymer, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1 , 3-dioxopentane, poly-1,3,6-trioxane, ethylene/maleic anhydride copolymer, polyamino acid (homopolymer or random copolymer), dextran or poly(n- vinylpyrrolidone) polyethylene glycol, propylene glycol homopolymer, propylene oxide/ethylene oxide copolymer, polyoxyethylated polyols (such as glycerol), polyvinyl alcohol, and mixtures thereof.
  • PEG polyethylene glycol
  • ethylene glycol/propylene glycol copolymer carboxymethylcellulose
  • dextran polyvinyl alcohol
  • polyvinylpyrrolidone poly-1 , 3-diox
  • Polyethylene glycol propionaldehyde offers advantages in manufacturing due to its stability in water.
  • the polymers can be of any molecular weight and can be branched or unbranched.
  • the number of polymers attached to the anti-C5a antibody can vary, and if more than one polymer is attached, they can be the same or different molecules.
  • the amount and/or type of polymer used for derivatization can be determined based on considerations including, but not limited to, the need to improve the properties or functionality of the anti-C5a antibody, whether the anti-C5a antibody derivative is useful under specific conditions. Treatment etc.
  • compositions comprising any of the anti-C5a antibodies (eg, full-length anti-C5a antibodies), nucleic acids encoding the antibodies, vectors comprising nucleic acids encoding the antibodies, or host cells comprising the nucleic acids or vectors described herein. substances, also referred to here as preparations).
  • a pharmaceutical composition comprising any of the anti-C5a antibodies described herein and a pharmaceutically acceptable carrier.
  • a suitable anti-C5a antibody can be obtained by mixing an anti-C5a antibody of the desired purity with an optional pharmaceutically acceptable carrier, excipient or stabilizer (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980))
  • Antibody preparations are prepared in the form of lyophilized preparations or liquid preparations.
  • Acceptable carriers, excipients or stabilizers are non-toxic to the recipient at the doses and concentrations used and include buffers such as phosphates, citric acid and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (e.g.
  • Octadecyldimethylbenzyl ammonium chloride hexamethylammonium chloride; benzalkonium chloride; benzethonium chloride; phenol; butanol or benzyl alcohol; alkyl parabens, such as p-hydroxybenzoate Methyl formate or propyl parahydroxybenzoate; catechol; resorcinol; cyclohexanol; 3-pentanol and m-cresol); low molecular weight (less than 10 residues) peptides; proteins, e.g.
  • hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal complexes such as zinc-protein complex); and/or non-ionic surfactants such as TWEEN TM , PLURONICS TM or polyethylene glycol (PEG); exemplary formulations are as described in WO98/56418, expressly incorporated herein by reference.
  • Lyophilized formulations suitable for subcutaneous administration are described in WO97/04801. Such lyophilized formulations can be reconstituted with appropriate diluents into high protein concentration formulations, and the reconstituted formulations can be administered subcutaneously to the individuals to be treated herein. Cationic liposomes or liposomes can be used to deliver the anti-C5a antibodies of the present application to cells.
  • the formulations described herein may also contain one or more other active substances necessary for the treatment of specific conditions, preferably substances with complementary activities and no adverse reactions to each other.
  • other active substances necessary for the treatment of specific conditions, preferably substances with complementary activities and no adverse reactions to each other.
  • it may be desirable to further include antineoplastic agents, growth inhibitory agents, cytotoxic agents, or chemotherapeutic agents. These molecules are present in combination and in amounts effective for the intended purpose. Effective amounts of other substances will depend on the amount of anti-C5a antibody in the formulation, the type of disease or condition or treatment, and other factors as discussed above.
  • These drugs are typically used at the same dosages and routes of administration as described herein, or at 1% to 99% of currently used dosages.
  • the anti-C5a antibodies can also be embedded in microcapsules prepared, for example, by coacervation techniques and interfacial polymerization, such as in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, respectively). , microemulsions, nanoparticles and nanocapsules) or in macroemulsions - microcapsules and poly(methyl methacrylate) microcapsules. Sustained release formulations can be prepared.
  • sustained release formulations of anti-C5a antibodies can be prepared.
  • suitable examples of sustained release formulations include solid hydrophobic polymeric semipermeable matrices containing the antibodies (or fragments thereof) in the form of shaped articles, for example, films or microcapsules.
  • sustained-release matrices include polyester, hydrogel (eg, poly(2-hydroxyethyl methacrylate) or poly(vinyl alcohol)), polylactic acid (U.S. Pat. No.
  • L-glutamine Acid and L-glutamic acid ethyl ester copolymer non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymer such as LUPRON DEPOTTM (a biodegradable product composed of lactic acid-glycolic acid copolymer and leuprolide acetate) injection microspheres) and poly-D(-)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic-glycolic acid can release molecules over 100 days, certain hydrogels can release proteins in much shorter time periods.
  • encapsulated antibodies When encapsulated antibodies stay in the body for a long time, they will denature or aggregate due to exposure to a humid environment at 37°C, which may lead to loss of biological activity or changes in immunogenicity.
  • Reasonable strategies can be designed to stabilize anti-C5a antibodies based on the corresponding mechanisms. For example, if the aggregation mechanism is found to be through thiodisulfide exchange to form intermolecular S-S bonds, this can be achieved by modifying sulfhydryl residues, lyophilizing in acidic solutions, controlling water content, using appropriate additives, and developing specific polymers. matrix composition to achieve stabilization.
  • the anti-C5a antibody (eg, full-length anti-C5a antibody) is formulated with citrate, sodium chloride, acetate, succinate, glycine, polysorbate 80 (Tween 80), or in any combination of the above buffers.
  • Preparations for in vivo administration must be sterile. This can be easily accomplished, for example, by applying sterile filtration membrane filtration.
  • Anti-C5a antibodies may be administered to an individual (e.g., a mammal, such as a human) to treat diseases and/or conditions resulting from dysregulation of the C5a signaling pathway (e.g., autoimmune and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune dysregulation diseases), including but not limited to inflammatory response syndrome (SIRS), sepsis , severe sepsis, septic shock, ischemia/reperfusion-related injury, acute lung injury, pneumonia, acute and chronic graft rejection in transplant patients, graft-versus-host reaction, glomerular disease, glomerulonephritis, renal failure of entities, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth and solid organ cancers.
  • SIRS inflammatory response syndrome
  • sepsis severe sepsis
  • septic shock ischemia/reper
  • the present application provides a method for treating diseases and/or conditions caused by dysregulation of the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplantation for immune dysregulation diseases), comprising administering to an individual an effective amount of a composition (e.g., a pharmaceutical composition) comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), such as any of the methods described herein.
  • a composition e.g., a pharmaceutical composition
  • an anti-C5a antibody e.g., a full-length anti-C5a antibody
  • the subject is a human.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a pharmaceutical composition comprising a C5a antibody (e.g., a full-length anti-C5a antibody) that specifically binds an epitope on human C5a, wherein the epitope Contains the amino acid residues of human C5a.
  • a pharmaceutical composition comprising a C5a antibody (e.g., a full-length anti-C5a antibody) that specifically binds an epitope on human C5a, wherein the epitope Contains the amino acid residues of human C5a.
  • the anti-C5a antibody is a full-length antibody.
  • the full-length anti-C5a antibody is an IgGl or IgG4 antibody.
  • the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, In the group consisting of Crohn's disease, tumor growth, and solid organ cancers.
  • the individual is a human.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 2, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 6, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 12.
  • a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1,
  • the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody.
  • the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
  • the individual is a human.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 25 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:25; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:35 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:35.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • disease comprising administering to the individual an effective amount of a composition
  • an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region.
  • the IgG1 is human IgG1.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 12.
  • a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1,
  • the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody.
  • the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic transplant rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
  • the individual is a human.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 26 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:26; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:36 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:36.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • disease comprising administering to the individual an effective amount of a composition
  • an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region.
  • the IgG1 is human IgG1.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 8, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 13.
  • a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1,
  • the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody.
  • the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
  • the individual is a human.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 27 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:27; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:37 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:37.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • disease comprising administering to the individual an effective amount of a composition
  • an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region.
  • the IgG1 is human IgG1.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 14.
  • a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1,
  • the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody.
  • the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
  • the individual is a human.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 28 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:28; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:38 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:38.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • disease comprising administering to the individual an effective amount of a composition
  • an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region.
  • the IgG1 is human IgG1.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 5, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 10, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 14.
  • a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1,
  • the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody.
  • the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic transplant rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
  • the individual is a human.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 29 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:29; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:39 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:39.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • disease comprising administering to the individual an effective amount of a composition
  • an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region.
  • the IgG1 is human IgG1.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 14.
  • a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1,
  • the anti-C5a antibody is a full-length antibody.
  • the full-length anti-C5a antibody is an IgGl or IgG4 antibody.
  • the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
  • the individual is a human.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 30 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:30; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:40 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:40.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • disease comprising administering to the individual an effective amount of a composition
  • an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region.
  • the IgG1 is human IgG1.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 12.
  • a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1,
  • the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody.
  • the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
  • the individual is a human.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 31 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:31; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:41 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:41.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • disease comprising administering to the individual an effective amount of a composition
  • an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region.
  • the IgG1 is human IgG1.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 15.
  • a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1,
  • the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody.
  • the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
  • the individual is a human.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 32 or Variants thereof, which variants have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:32; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:42 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:42.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • disease comprising administering to the individual an effective amount of
  • an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region.
  • the IgG1 is human IgG1.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 14.
  • a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1,
  • the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody.
  • the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic transplant rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
  • the individual is a human.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 33 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:33; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:43 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:43.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • disease comprising administering to the individual an effective amount of a composition
  • an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region.
  • the IgG1 is human IgG1.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 11, or the Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 16.
  • a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1,
  • the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody.
  • the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
  • the individual is a human.
  • a method for treating diseases associated with the C5a signaling pathway comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 34 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:34; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:44 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:44.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • diseases associated with the C5a signaling pathway e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • disease comprising administering to the individual an effective amount of a composition
  • an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region.
  • the IgG1 is human IgG1.
  • the IgG4 is human IgG4.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45.
  • the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47.
  • the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
  • the subject is a mammal (eg, human, non-human primate, rat, mouse, cow, horse, porcine, sheep, goat, dog, cat, etc.).
  • the individual is a human.
  • the individual is a clinical patient, clinical trial volunteer, experimental animal, etc.
  • the individual is less than 60 years old (including, for example, less than 50, 40, 30, 25, 20, 15, or 10 years old).
  • the individual is older than 60 years old (including, for example, older than 70, 80, 90, or 100 years old).
  • the individual is diagnosed with or genetically predisposed to one or more diseases or conditions described herein (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or or pain and/or respiratory and/or transplant immune disorders).
  • the individual has one or more risk factors associated with one or more diseases or conditions described herein.
  • the application provides a method of delivering an anti-C5a antibody (e.g., any of the anti-C5a antibodies described herein, e.g., an isolated anti-C5a antibody) to cells in an individual that express C5a on their surface, said The method includes administering to the individual a composition comprising an anti-C5a antibody.
  • an anti-C5a antibody e.g., any of the anti-C5a antibodies described herein, e.g., an isolated anti-C5a antibody
  • autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune dysregulation diseases or any other diseases showing aberrant expression of C5a and clinical description of these diseases are known in the art. Such methods include, but are not limited to, immunohistochemistry, PCR, and fluorescence in situ hybridization (FISH), for example.
  • FISH fluorescence in situ hybridization
  • the anti-C5a antibodies e.g., full-length anti-C5a antibodies
  • compositions described herein are combined with a second, third, or fourth agent (including, for example, an anti-tumor agent, a growth inhibitory agent, a cytotoxic agent or chemotherapeutic agents) to treat diseases related to the C5a signaling pathway.
  • a second, third, or fourth agent including, for example, an anti-tumor agent, a growth inhibitory agent, a cytotoxic agent or chemotherapeutic agents
  • Cancer treatments are evaluated using, for example, tumor regression, reduction in tumor weight or size, time to progression, survival, progression-free survival, overall response rate, duration of response, quality of life, protein expression levels and/or activity.
  • Methods of determining the effectiveness of treatment may be employed, including, for example, detection of response by radiographic imaging.
  • the effect of treatment is evaluated as percent tumor growth inhibition (%TGI), calculated using the equation 100 - (T/C ⁇ 100), where T is the relative mean tumor volume of the treated tumor and C is Relative mean tumor volume of untreated tumors.
  • %TGI percent tumor growth inhibition
  • the %TGI is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95% or more than 95%.
  • the therapeutic effect is assessed by changes in granulocyte morphology and/or an increase in the number of viable granulocytes.
  • therapeutic efficacy is assessed by increased secretion of cytokines by monocytes.
  • an anti-C5a antibody eg, isolated anti-C5a antibody
  • the dosage of an anti-C5a antibody composition administered to an individual may vary depending on the particular composition, the mode of administration, and the type of disease being treated.
  • the amount of the composition e.g., a composition comprising an anti-C5a antibody
  • the amount of anti-C5a antibody composition is sufficient to produce a complete response in an individual.
  • the amount of anti-C5a antibody composition is sufficient to produce a partial response in an individual.
  • the anti-C5a antibody composition is administered at a dose (e.g., when administered alone) sufficient to produce greater than 20%, 25%, 30%, 35%, or more in a population of individuals treated with the anti-C5a antibody composition.
  • An individual's response to the treatment methods described herein can be determined, for example, by the level of RECIST.
  • the amount of the composition is sufficient to prolong progression-free survival in an individual. In some embodiments, the amount of the composition is sufficient to prolong the overall survival of the subject. In some embodiments, the amount of the composition (eg, when administered alone) is sufficient to produce greater than 50%, 60%, 70%, or 77% of clinical benefit in a population of individuals treated with an anti-C5a antibody composition.
  • the amount of a composition is before treatment or with An amount sufficient to control symptoms and reduce the risk of exacerbation compared to the corresponding activity in other subjects not receiving treatment.
  • the magnitude of this effect can be measured using standard methods, such as in vitro assays of purified enzymes, cell-based assays, animal models, or human trials.
  • the amount of anti-C5a antibody (e.g., full-length anti-C5a antibody) in the composition is less than the amount that causes a toxic effect (i.e., an amount that is greater than a clinically acceptable level of toxicity) when the composition is administered to an individual. effect), or at a level at which potential side effects can be controlled or tolerated.
  • a toxic effect i.e., an amount that is greater than a clinically acceptable level of toxicity
  • the amount of the composition approximates the maximum tolerated dose (MTD) of the composition. In some embodiments, the amount of the composition is greater than 80%, 90%, 95%, or 98% of the MTD.
  • the amount of anti-C5a antibody (eg, full-length anti-C5a antibody) in the composition ranges from 0.001 ⁇ g to 1000 ⁇ g.
  • the effective amount of anti-C5a antibody eg, full-length anti-C5a antibody in the composition ranges from 0.1 ⁇ g/kg to 100 mg/kg based on body weight.
  • Anti-C5a antibody compositions can be administered to an individual (e.g., a human) by a variety of routes, including, for example, intravenous injection, intraarterial administration, intraperitoneal injection, intrapulmonary administration, oral administration, inhalation administration, intravascular administration, Intramuscular, intratracheal, subcutaneous, intraocular, intrathecal, mucosal or transdermal administration.
  • a sustained release formulation of the composition is used.
  • the composition is administered intravenously.
  • the composition is administered intraarterially.
  • the composition is administered intraperitoneally.
  • the composition is administered intrahepatically.
  • the composition is administered by hepatic artery infusion.
  • the composition is administered to a site distal to the first lesion.
  • an article of manufacture comprising a substance that can be used to treat diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and or cancer and/or pain and/or respiratory and/or transplant immune disorders), or for delivering an anti-C5a antibody (e.g., a full-length anti-C5a antibody) to cells expressing C5a on their surface.
  • the article of manufacture may include a container and a label or package insert on or accompanying the container. Suitable containers include, for example, bottles, vials, syringes, and the like. Containers can be made from a variety of materials, such as glass or plastic.
  • the container contains a composition effective for treating the disease or condition described herein and has a sterile port (eg, the container may be an IV bag or a vial with a hypodermic needle-pierceable cap). At least one active substance in the composition is the anti-C5a antibody described in this application.
  • the label or package insert identifies the specific condition for which the composition is used to treat.
  • the label or package insert further contains instructions for administering the anti-C5a antibody composition to a patient. Articles and kits including combination therapies are considered within the scope of this article.
  • Package insert means the instructions usually included in the commercial packaging of therapeutic products that contain information on indications, usage, dosage, administration, contraindications and/or warnings relevant to the use of these therapeutic products.
  • the package insert indicates that the composition can be used to treat diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or Transplant immune disorders).
  • the package insert states that the composition can be used to treat conditions including inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, Pneumonia, acute and chronic graft rejection in transplant patients, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases , inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
  • SIRS inflammatory response syndrome
  • the article of manufacture may further include a second container containing a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffer, Green's solution, or dextrose solution.
  • a pharmaceutically acceptable buffer such as bacteriostatic water for injection (BWFI), phosphate buffer, Green's solution, or dextrose solution.
  • kits that can be used for various purposes, for example for the treatment of diseases associated with the C5a signaling pathway (e.g. autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or immune disorders), or for delivering anti-C5a antibodies (eg, full-length anti-C5a antibodies) to cells expressing C5a on their surface, optionally in combination with an article of manufacture.
  • Kits of the present application include one or more containers comprising an anti-C5a antibody composition (or single dose form and/or article of manufacture) and, in some embodiments, further comprising another agent (e.g., an agent described herein ) and/or instructions for use consistent with any method described herein.
  • the kit may further include instructions for selecting individuals suitable for treatment. Instructions for use accompanying the kit in this application are usually written instructions on the label or package insert (e.g., a sheet of paper included in the kit), machine-readable instructions (e.g., instructions on a magnetic or optical storage disc) Also acceptable.
  • the kit includes a composition comprising an anti-C5a antibody (eg, a full-length anti-C5a antibody).
  • a kit includes: a) a composition comprising any one of the anti-C5a antibodies described herein, and b) an effective amount of at least one other agent capable of enhancing the effects of the anti-C5a antibody (e.g., treating effect, detection effect).
  • a kit includes: a) a composition comprising any one of the anti-C5a antibodies described herein, and b) administering the anti-C5a antibody composition to an individual for treating a disease associated with the C5a signaling pathway (e.g., Instructions for use in autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders).
  • a disease associated with the C5a signaling pathway e.g., Instructions for use in autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • a kit includes: a) a composition comprising any one of the anti-C5a antibodies described herein, and b) an effective amount of at least one other agent capable of enhancing the effects of the anti-C5a antibody (e.g., treating effect, detecting effect) and c) administering anti-C5a antibody compositions and other substances to an individual for the treatment of diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and /or respiratory and/or transplant immune disorders) instructions for use.
  • the anti-C5a antibody and other substances may be present in separate containers or in the same container.
  • the kit may include one specific composition or two or more compositions, one of which includes an anti-C5a antibody and another of which includes another agent.
  • the kit includes a nucleic acid (or a set of nucleic acids) encoding an anti-C5a antibody (eg, a full-length anti-C5a antibody).
  • the kit comprises: a) a nucleic acid (or set of nucleic acids) encoding an anti-C5a antibody (eg, a full-length anti-C5a antibody), and b) a host expressing the nucleic acid (or set of nucleic acids) cell.
  • the kit comprises: a) a nucleic acid (or a set of) nucleic acids encoding an anti-C5a antibody (e.g., a full-length anti-C5a antibody), and b) instructions for: i) in a host cell Expressing an anti-C5a antibody, ii) preparing a composition comprising the anti-C5a antibody, and iii) administering the composition comprising the anti-C5a antibody to an individual to treat a disease associated with the C5a signaling pathway (e.g., autoimmune disease and/or inflammatory disorder and/or cancer and/or pain and/or respiratory and/or transplant immune disorders).
  • a disease associated with the C5a signaling pathway e.g., autoimmune disease and/or inflammatory disorder and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • the kit includes: a) a nucleic acid (or a set of nucleic acids) encoding an anti-C5a antibody (eg, a full-length anti-C5a antibody), b) a host cell expressing the nucleic acid (or a set of nucleic acids) , and c) instructions for use in: i) expressing an anti-C5a antibody in a host cell, ii) preparing a composition comprising an anti-C5a antibody, and iii) administering a composition comprising an anti-C5a antibody to an individual to treat patients with C5a signaling Pathway related diseases (eg autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplant immune disorders).
  • C5a signaling Pathway related diseases eg autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplant immune disorders.
  • kits for detection and/or diagnosis comprising any of the anti-C5a antibodies described herein.
  • the kit includes a conjugate comprising any anti-C5a antibody described herein and a coupling moiety, wherein the coupling moiety is a radioactive isotope, a fluorescent substance, a luminescent substance , colored substances or enzymes.
  • it relates to the use of any of the anti-C5a antibodies described in this application or the conjugate of the invention in preparing a kit for detecting the presence or level of C5a in a sample.
  • an in vitro method of detecting the presence or level of C5a in a sample comprising: a) contacting the sample with any of the methods described herein under conditions that allow the formation of a complex between the antibody and C5a. contacting an anti-C5a antibody or conjugate; and b) detecting the formation of a complex; and c) determining the presence or level of C5a in the sample.
  • a method for diagnosing abnormalities in C5a levels comprising: a) obtaining a sample to be tested, and b) combining the sample to be tested with the method described herein under conditions that allow the formation of a complex between the antibody and C5a. Contact any of the anti-C5a antibodies or conjugates; c) detect the formation of the complex; d) determine the level of C5a in the sample to be tested; and e) determine whether the level of C5a in the sample to be tested is higher than normal Levels of C5a in group samples.
  • the sample is a serum sample or plasma sample.
  • the sample to be tested is derived from a patient in need.
  • the patient has or has not received C5a antibody treatment.
  • kits described in this application are packaged in a suitable form.
  • suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (eg, sealed mylar or plastic bags), and the like. Kits may optionally provide other components such as buffers and instructions. Accordingly, the present application also provides articles including vials, bottles, jars, flexible packaging (eg, sealed Mylar or plastic bags), and the like.
  • Containers may be unit dose, bulk (eg, multi-dose packaging) or subunit dose.
  • a kit is provided that contains a sufficient dose of an anti-C5a antibody as described herein (e.g., a full-length anti-C5a antibody) to effectively treat an individual for a long period of time, such as one week, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months or longer time.
  • the kit may also contain multiple unit doses of the anti-C5a antibody, the pharmaceutical composition and instructions for use, and be packaged in quantities sufficient for storage and use in pharmacies, such as hospital pharmacies and compounding pharmacies.
  • complement component 5a complement component 5a, C5a
  • Bavih-C5a Biotin-Avi-10his-C5a
  • Example 1 Preparation of recombinant human C5a and screening of anti-C5a single chain antibodies (scFv)
  • the cDNA encoding human C5a (synthesized by Shanghai Jierui Bioengineering Co., Ltd.) was constructed into the prokaryotic expression vector pTWIN1 or the eukaryotic expression vector pTT5 by subcloning. Add His tag or other tags commonly used by those skilled in the art at the end of the cDNA encoding the above protein.
  • the pTWIN1-C5a and pTT5-Avi-10his-C5a expression vectors were constructed and produced. Among them, “his” or “His” represents the His tag, and "Avi” represents the avidin tag.
  • the eukaryotic expression vector pTT5-Avi-10his-C5a was transfected into 293F cells, and the above cells were cultured at 37°C, 8% CO 2 , and 120 rpm for 5 days. Collect cell culture fluid. According to the operating instructions, a nickel column (Ni) was used to purify the protein expressing His tag. The specific operation is as follows: Immobilized metal affinity chromatography (IMAC) is performed using Ni-NTA from QIAGEN Company.
  • IMAC Immobilized metal affinity chromatography
  • buffer A1 50mM Na 3 PO 4 , 0.15M NaCl, pH 7.2
  • the pH of the culture supernatant was adjusted to 7.2, and the sample was loaded at room temperature with a flow rate of 150cm/h.
  • the column was equilibrated again with 6 column volumes of A1 buffer at a flow rate of 150 cm/h.
  • 10 times the column volume of 50mM PB solution (containing 0.15M NaCl and 0.2M imidazole, pH 7.2) was used for elution, and the eluent was collected.
  • the recombinant human C5a prokaryotic expression vector pTWIN1-C5a was transformed into E. coli and expanded cultured. After collecting the bacterial cells, add 300ml of 50mM Tris solution, stir to dissolve, and place in a -40°C refrigerator overnight. The next day, the above samples were taken out and thawed, and the cells were disrupted using an ultrasonic disruptor. After centrifugation, take the supernatant and filter the supernatant with a 0.8um filter membrane.
  • Avi-10His-C5a was biotinylated using biotinylated ligase B0101A (GeneCopoeia). Briefly, BufferA/B and BirA ligase were added to Avi-10His-C5a and incubated at 30°C for 2 hours. Biotinylated Avi-10His-C5a was named Bavih-C5a. Biotinylation efficiency was detected by ELISA method. Briefly, the starting concentration of Bavih-C5a was set to 500ng/ml, diluted at a ratio of 1:2, and then coated on the ELISA plate after dilution. SA-HRP was used to detect the signal, and biotinylated standards were used as controls. The Bavih-C5a biotinylation labeling efficiency was determined to be 70%.
  • Construct scFv antibody phage display library use recombinant human C5a as the antigen, immunize rabbits together with adjuvant, collect the animal serum after immunization, and detect the total IgG titer in the rabbit serum after immunization by ELISA. After several rounds of immunization, peripheral blood, lymph nodes, and spleen were used to establish a phage display library. Briefly, peripheral blood, lymph nodes and spleens of immunized rabbits were collected, RNA was extracted, and cDNA was obtained by reverse transcription. VH and VK fragments were amplified using VH and VK specific primers.
  • VH and VK were ligated to construct scFv and cloned into the phage display plasmid pDAN5. Subsequently, the plasmid was electroporated into E. coli TG1, and the phage was used to infect E. coli TG1 to obtain a scFv antibody phage display library.
  • scFv single chain antibodies
  • 2x10 11 PFU of the phage scFv library was added to Bavih-C5a and incubated at 37°C for 2 hours. Phage that bind to human C5a are captured by streptavidin-coated magnetic beads, while unbound phage are washed away. After washing 8-15 times with TBST solution (as the number of screening rounds increases, the number of washes increases), Glycine-HCl solution (pH 2.2) is used to elute the phage that specifically binds human C5a.
  • the obtained positive scFv antibody is reconstituted into a human IgG1 or IgG4 antibody molecule having a heavy chain constant region of human IgG1 or IgG4 and a human kappa or lambda light chain constant region.
  • V L and V H were amplified from the phage expression vector and constructed into the eukaryotic expression vector pTT5-K (containing the kappa constant region) or pTT5-L (containing the lambda constant region) and pTT5-H1 (containing the IgG1 heavy chain constant region) respectively. ) or pTT5-H4 (containing the IgG4 heavy chain constant region).
  • the plasmid expressing the light chain and the plasmid expressing the heavy chain were co-transfected into 293F cells. Cultivate for 5 days at 37°C, 8% CO 2 , 120 rpm, and purify the culture solution using a Protein A affinity chromatography column. Briefly, the Protein A column was first equilibrated with 6 column volumes of PBS buffer (containing 50 mM PBS and 0.15 M NaCl, pH 7.2) at a flow rate of 150 cm/h. The pH of the culture supernatant was adjusted to 7.2, and the sample was loaded at room temperature with a flow rate of 150cm/h. After complete equilibrium, 50 mM sodium citrate buffer (pH 3.5) was added for elution, and the eluate was collected.
  • PBS buffer containing 50 mM PBS and 0.15 M NaCl, pH 7.2
  • Each antibody sample was first diluted to 10 ⁇ g/mL, followed by gradient dilution in a 1:3 ratio. Add the gradient diluted samples to a 96-well plate, 100 ⁇ L per well, and incubate at 37°C for 1 hour. Then wash 6 times with TBST solution, add goat anti-human IgG Fc-AP antibody (SouthernBiotech, 2048-04; 1:2500), 100 ⁇ L per well, and incubate at 37°C for 1 hour. After washing with TBST plate washer 6 times, add 50 ⁇ L of pNPP to each well, incubate at 37°C for 10 min, and terminate the reaction with 3M NaOH. The absorbance value at 410 nm was read and a binding curve was generated by PRISM.
  • each antibody sample was first diluted to 10 ⁇ g/mL, followed by gradient dilution in a 1:3 ratio. Add the gradient diluted samples to a 96-well plate, 100 ⁇ L per well, and incubate at 37°C for 1 hour. Then wash with TBST solution 6 times, add anti-human Fc-AP antibody (1:2500), 100 ⁇ L per well, and incubate at 37°C for 1 hour.
  • ROS Reactive oxygen species
  • C5a can stimulate neutrophils to release reactive oxygen species (ROS), thereby promoting neutrophils to participate in a wide range of inflammatory responses. Based on this mechanism, induced neutrophils were used to detect the blocking effect of anti-C5a antibodies.
  • the HL-60 cell line is human promyelocytic leukemia cells. Briefly, HL60 cells were treated with 1mM dibutyryl cAMP sodium salt (sigma, D0260) for 48 hours to induce their differentiation. The cells shrank to a spindle shape and became neutral. Granulocytic differentiation. C5a stimulates ROS production in differentiated HL-60 cells in a dose-dependent manner.
  • ROS reactive oxygen species
  • the inhibition rates with only C5a stimulation and without C5a stimulation were calculated as 0% and 100%, respectively.
  • the experimental data were normalized to calculate the IC 50 value of the C5a antibody.
  • Anti-C5a antibodies RH23, RH25, RH32, RH35, RH36, RH58, RH85, RH86, RH92, and RH95 (reconstituted in the form of adult IgG4) can all inhibit ROS release well and are comparable to the control antibody INab308. .
  • Example 3 In vivo effect of anti-C5a antibody in treating ARDS caused by coronavirus
  • ARDS animal disease model and normal control group A total of 46 C5a humanized mice (purchased from Shanghai Southern Model Biotechnology Co., Ltd.) were used. 40 of them were administered 4, 3 and 2 days before the experiment.
  • mice were injected with adenovirus carrying and expressing SARS-CoV-2 N protein (see Ting Gao et al., https://doi.org/10.1101/2020.03.29.20041962), 7.5 ⁇ 10 8 PFU/100 ⁇ L/mouse/time/ day, the disease model groups were divided into model control group, high-dose group (10mg/kg), medium-dose group (3mg/kg) and low-dose group (1mg/kg). On day 0 (the day of the experiment), mice were injected with the corresponding doses of antibodies in parentheses. The remaining 6 mice were injected with 100 ⁇ L of 0.9% sodium chloride solution as a normal control group.
  • mice in the disease model control group and each experimental group were injected with LPS-K235 (Sigma-Aldrich) at a concentration of 1 mg/mL, 100 ⁇ L/mouse. All reagents in this study were administered by tail vein injection. The survival of mice in each group was observed and analyzed at 12h, 24h, 36h, 48h, 60h and 72h after administration. 72 hours after antibody administration, the mice were anesthetized, and blood was collected from the orbits. use A series of blood analyzers perform whole blood white blood cell counting and classification, including white blood cell count (WBC), neutrophils (Neut), lymphocytes (Lymph), and monocytes (Mono). ELISA method was used to detect the levels of GM-CSF, IL-1 ⁇ , IL-6, TNF- ⁇ , MCP-1 and other cytokines.
  • the anti-C5a antibody RH86 of the present application can effectively reduce or prevent the death of mice caused by coronavirus and improve the survival rate of mice.
  • the levels of WBC, Lymph and Mono of mice in the model control group were reduced, and the levels of GM-CSF, IL-1 ⁇ , IL-6, TNF- ⁇ , and MCP-1 were significantly increased. There is a statistically significant difference between them (P ⁇ 0.05).
  • the three dose experimental groups administered anti-C5a antibody (RH86) all showed an increase in the number of WBC and Lymph, GM-CSF, IL-1 ⁇ , IL-6, TNF- ⁇ , MCP- 1.
  • C5a levels decreased in a dose-dependent manner.

Abstract

The present invention relates to an antibody specifically recognizing C5a. The antibody has high binding activity to human C5a or human C5a-desArg. Moreover, the present invention further relates to a nucleotide sequence encoding the antibody, a vector and a cell comprising the nucleotide sequence, a pharmaceutical composition comprising the antibody, a use of the antibody in the preparation of a drug for treating autoimmune diseases and inflammation, cancer, pain, respiration and/or transplantation-related diseases, and a use of the antibody or a conjugate thereof in the preparation of a kit for detecting the presence or level of C5a.

Description

特异性识别C5A的抗体及其应用Antibodies that specifically recognize C5A and their applications
相关申请的交叉引用Cross-references to related applications
[根据细则91更正 11.07.2023]
本申请要求申请号为202210350226.9，申请日为2022.04.02，发明名称为“特异性识别C5A的抗体及其应用”的中国专利申请的优先权，且该申请的全部内容以引用方式并入本文中。[Correction 11.07.2023 under Rule 91]
This application claims priority to the Chinese patent application with application number 202210350226.9, filing date 2022.04.02, and the invention title is "Antibody that specifically recognizes C5A and its application", and the entire content of this application is incorporated herein by reference. .
对电子序列表的引用Reference to electronic sequence listing
电子序列表(文本名称：CN_202203244652.xml，记录日期：2022.03.24，大小：141KB)的内容通过整体引用并入本文中。The contents of the electronic sequence listing (text name: CN_202203244652.xml, recording date: 2022.03.24, size: 141KB) are incorporated into this article by reference in its entirety.
技术领域Technical field
本申请涉及特异性识别补体组分5a(C5a)的抗体及其制备方法和用途，包括用于治疗自身免疫性疾病、炎症、癌症、疼痛、呼吸和/或移植相关疾病的用途。The present application relates to antibodies that specifically recognize complement component 5a (C5a) and their preparation methods and uses, including uses for the treatment of autoimmune diseases, inflammation, cancer, pain, respiratory and/or transplantation-related diseases.
背景技术Background technique
C5a是过敏性反应和炎症过程中的一种活性肽，其在补体级联反应中由C5转化酶切割补体组分C5而形成。C5a刺激肥大细胞脱粒、肿瘤坏死因子-α(TNF-α)和组胺的释放，还通过增加内皮细胞表面粘附分子的表达从而将吞噬细胞募集至感染和炎症部位(Mollnes,T.E.et al.Blood 2002,100,1869–1877；Riedemann,N.C.et al.Immunity 2003,19,193–202)。在一些病理刺激的情况下，如移植后异体移植排斥和哮喘，C5a还导致血管通透性增加(Gueler,F.et al.J.Am.Soc.Nephrol.2008,19,2302–2312；Krug,N.et al.Am.J.Respir.Crit.Care Med.2001,164,1841-1843；Khan,M.A.et al.Proc.Natl.Acad.Sci.USA 2013,110,6061–6066)。已有多项研究讨论过血清中C5a水平。在Lechner等人的一项研究中，对照组中C5a的水平为8.34+2.05(ng/mL)(Lechner,J.et al.Immun.Ageing2016,13,4)。另有研究表明，在正常情况下，由于过敏毒素的快速清除，血浆中C5a水平非常低(Oppermann,M.et al.Immunology 1994,82,516–521)。使用C5a(25nM)处理后，小鼠皮质肾小管细胞中转化生长因子-β(TGF-β)的水平升高，这表明C5a可导致肾纤维化和肾疤痕形成(Boor,P.et al.J.Am.Soc.Nephrol.2007,18,1508–1515)。C5a is an active peptide in allergic reactions and inflammation, which is formed during the complement cascade by cleavage of complement component C5 by C5 convertase. C5a stimulates mast cell degranulation, tumor necrosis factor-α (TNF-α) and histamine release, and also recruits phagocytes to sites of infection and inflammation by increasing the expression of adhesion molecules on the surface of endothelial cells (Mollnes, T.E. et al. Blood 2002, 100, 1869–1877; Riedemann, N.C. et al. Immunity 2003, 19, 193–202). In some cases of pathological stimulation, such as post-transplant allograft rejection and asthma, C5a also leads to increased vascular permeability (Gueler, F. et al. J. Am. Soc. Nephrol. 2008, 19, 2302–2312; Krug ,N.et al.Am.J.Respir.Crit.Care Med.2001,164,1841-1843; Khan,M.A.et al.Proc.Natl.Acad.Sci.USA 2013,110,6061–6066). Several studies have discussed C5a levels in serum. In a study by Lechner et al., the level of C5a in the control group was 8.34+2.05 (ng/mL) (Lechner, J. et al. Immun. Ageing 2016, 13, 4). Other studies have shown that under normal circumstances, C5a levels in plasma are very low due to the rapid clearance of anaphylatoxins (Oppermann, M. et al. Immunology 1994, 82, 516–521). After treatment with C5a (25nM), the levels of transforming growth factor-β (TGF-β) in mouse cortical tubular cells increased, indicating that C5a can cause renal fibrosis and renal scarring (Boor, P. et al. J. Am. Soc. Nephrol. 2007, 18, 1508–1515).
C5a是一种有效的促炎分子，与一种经典的G蛋白偶联受体(GPCR)C5aRI(CD88)结合并引发促炎信号通路激活(Li,R.et al.FASEB J.2013,27,855–864)。C5aR在非髓样细胞上广泛表达，如脐血管内皮细胞(HUVEC)，鼠类真皮、肝、肺和肾近端小管(Monsinjon,T.et al.FASEB J.2003,17,1003–1014；Gerard,C.et al.Annu.Rev.Immunol.1994,12,775–808；Haviland,D.L.et al.J.Immunol.1995,154,1861–1869)。此外，研究证明C5aR在肾小球内皮细胞而非足细胞中表达，这表明C5a可能主要在肾内皮细胞中引起蛋白尿(Tsai,I.J.et al.Cell.Mol.Life Sci.2015,72,3157–3171)。C5a is a potent pro-inflammatory molecule that binds to a classic G protein-coupled receptor (GPCR) C5aRI (CD88) and triggers the activation of pro-inflammatory signaling pathways (Li, R. et al. FASEB J. 2013, 27, 855 –864). C5aR is widely expressed on non-myeloid cells, such as umbilical vascular endothelial cells (HUVEC), murine dermis, liver, lung and renal proximal tubules (Monsinjon, T. et al. FASEB J. 2003, 17, 1003–1014; Gerard, C. et al. Annu. Rev. Immunol. 1994, 12, 775–808; Haviland, D. L. et al. J. Immunol. 1995, 154, 1861–1869). In addition, studies have demonstrated that C5aR is expressed in glomerular endothelial cells rather than podocytes, suggesting that C5a may mainly cause proteinuria in renal endothelial cells (Tsai, I.J. et al. Cell. Mol. Life Sci. 2015, 72, 3157 –3171).
因此，通过中和C5a阻断其与C5aR的结合，成为一种治疗C5a介导的疾病和病症的方法。专利申请WO2011063980中披露了针对人C5a的抗体INab308(InflaRx)，WO2012088247中披露了C5a抗体MEDI-7814(MedImmune)，US10450370中披露了C5a抗体BNJ383(Alexion)。Therefore, blocking its binding to the C5aR by neutralizing C5a emerges as an approach to treat C5a-mediated diseases and conditions. Patent application WO2011063980 discloses the antibody INab308 (InflaRx) against human C5a, WO2012088247 discloses the C5a antibody MEDI-7814 (MedImmune), and US10450370 discloses the C5a antibody BNJ383 (Alexion).
本文提及的所有出版物、专利、专利申请和已公开的专利申请中披露的内容，以引用方式全部并入本文中。The disclosures in all publications, patents, patent applications, and published patent applications mentioned herein are hereby incorporated by reference in their entirety.
申请概述Application Overview
在一些实施例中，提供一种分离的抗C5a抗体，其包含：(i)VH，其包含如氨基酸序列SEQ ID NO:25所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:35所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；(ii)VH，其包含如氨基酸序列SEQ ID NO:26所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:36所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；(iii)VH，其包含如氨基酸序列SEQ ID NO:27所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:37所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；(iv)VH，其包含如氨基酸序列SEQ ID NO:28所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:38所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；(v)VH，其包含如氨基酸序列SEQ ID NO:29所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:39所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；(vi)VH，其包含如氨基酸序列SEQ ID NO:30所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:40所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；(vii)VH，其包含如氨基酸序列SEQ ID NO:31所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:41所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；(viii)VH，其包含如氨基酸序列SEQ ID NO:32所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:42所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；(ix)VH，其包含如氨基酸序列SEQ ID NO:33所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:43所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；或(x)VH，其包含如氨基酸序列SEQ ID NO:34所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:44所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3。 In some embodiments, an isolated anti-C5a antibody is provided, comprising: (i) a V H comprising HC-CDR1, HC-CDR2 and HC as shown in the amino acid sequence of SEQ ID NO:25 -CDR3; and VL , which comprises LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO:35; (ii) VH , which comprises the amino acid sequence SEQ ID NO V H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as shown in: 26; and V L comprising LC-CDR1, LC-CDR2 and V L as shown in amino acid sequence SEQ ID NO: 36 LC-CDR3; (iii) VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO:27; and VL comprising the amino acid sequence SEQ ID LC-CDR1, LC-CDR2 and LC-CDR3 included in the V L shown in NO: 37; (iv) V H including HC-CDR1, HC included in the V H shown in the amino acid sequence SEQ ID NO: 28 -CDR2 and HC-CDR3; and VL comprising LC-CDR1, LC - CDR2 and LC-CDR3 as shown in the amino acid sequence SEQ ID NO:38; (v) VH comprising amino acids such as HC-CDR1, HC-CDR2 and HC-CDR3 included in VH shown in the sequence SEQ ID NO:29; and VL including LC-CDR1, LC-CDR2 and LC-CDR3; (vi) VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO:30; and VL comprising as LC-CDR1, LC-CDR2 and LC-CDR3 included in the V L shown in the amino acid sequence SEQ ID NO: 40; (vii) V H including HC included in the V H shown in the amino acid sequence SEQ ID NO: 31 -CDR1, HC-CDR2 and HC-CDR3; and VL comprising LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO:41; (viii) VH , It contains HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO:32; and VL , which comprises VL comprised as shown in the amino acid sequence SEQ ID NO:42 LC-CDR1, LC-CDR2 and LC-CDR3; (ix) VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO:33; and VL , which includes LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO:43; or (x) VH , which includes the LC-CDR1, LC-CDR2 and LC-CDR3 as shown in the amino acid sequence SEQ ID NO:34 VH comprises HC-CDR1, HC-CDR2 and HC-CDR3; and VL comprises VL comprising LC-CDR1, LC-CDR2 and LC-CDR3 as shown in the amino acid sequence of SEQ ID NO:44.
在一些实施例中，提供一种分离的抗C5a抗体，其包含：(i)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:2，和HC-CDR3，其包含氨基酸序列SEQ ID NO:6，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(ii)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(iii)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:8，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:13，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:21，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(iv)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(v)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:5，和HC-CDR3，其包含氨基酸序列SEQ ID NO:10，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(vi)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(vii)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:23，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(viii)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:15，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(ix)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；或(x)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:11，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:16，LC-CDR2，其包含氨基酸序列SEQ ID NO:19，和LC-CDR3，其包含氨基酸序列SEQ ID NO:24，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。In some embodiments, an isolated anti-C5a antibody is provided, comprising: (i) V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising and V L , The V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 20, or Variants of the V L , including substitutions of up to about 5 amino acids in its LC-CDRs; (ii) V H , the V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC- CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H whose HC-CDRs comprise substitutions of up to about 5 amino acids; and VL , said VL comprising: LC-CDR1, which comprises the amino acid sequence SEQ ID NO: 12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which comprises the amino acid sequence SEQ ID NO :20, or a variant of the VL , comprising at most about 5 amino acid substitutions in its LC-CDRs; (iii) VH , the VH comprising: HC-CDR1, comprising the amino acid sequence SEQ ID NO: 1. HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 8, or a variant of the V H , which HC-CDRs include up to about 5 Substitutions of amino acids; and V L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO: 13, LC-CDR2 comprising the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO: 17 Sequence SEQ ID NO: 21, or a variant of the V L containing up to about 5 amino acid substitutions in its LC-CDRs; (iv) V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9, or a variant of the V H , whose HC-CDRs include Substitutions of up to about 5 amino acids; and V L comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 14, LC-CDR2 comprising the amino acid sequence SEQ ID NO: 17, and LC-CDR3 , which includes the amino acid sequence SEQ ID NO: 22, or a variant of the V L , whose LC-CDRs include substitutions of up to about 5 amino acids; (v) V H , the V H includes: HC-CDR1, It comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 5, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 10, or a variant of said V H whose HC - CDRs comprising substitutions of up to about 5 amino acids; and V L comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 14, LC-CDR2 comprising the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said VL , whose LC-CDRs comprise up to about 5 amino acid substitutions; (vi) VH , said VH comprising: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7, or a variant of the V H A body comprising up to about 5 amino acid substitutions in its HC-CDRs; and a VL comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO: 14, LC-CDR2 comprising the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , which contains up to about 5 amino acid substitutions in its LC-CDRs; (vii) V H , said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and VL comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 12, LC-CDR2 comprising The amino acid sequence SEQ ID NO: 17, and LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 23, or a variant of the V L containing up to about 5 amino acid substitutions in its LC-CDRs; (viii) V H , the V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7 , or a variant of the V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 15, LC- CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of the V L whose LC-CDRs comprise substitutions of up to about 5 amino acids; (ix) V H comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO: 14. LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , which contains up to about 5 LC-CDRs Substitution of amino acids; or (x) V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, and HC-CDR3, It comprises the amino acid sequence SEQ ID NO: 11, or a variant of said V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC-CDR1 comprising amino acids Sequence SEQ ID NO: 16, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 19, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 24, or a variant of the V L in which LC-CDRs Contains substitutions of up to about 5 amino acids.
在一些实施例中，如上所述任一种分离的抗C5a抗体，其包含：(i)VH，其包含SEQ ID NO:25所示的氨基酸序列或其变体，所述变体与SEQ ID NO:25所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:35所示的氨基酸序列或其变体，所述变体与SEQ ID NO:35所示的氨基酸序列具有至少约80％序列同一性；(ii)VH，其包含SEQ ID NO:26所示的氨基酸序列或其变体，所述变体与SEQ ID NO:26所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:36所示的氨基酸序列或其变体，所述变体与SEQ ID NO:36所示的氨基酸序列具有至少约80％序列同一性；(iii)VH，其包含SEQ ID NO:27所示的氨基酸序列或其变体，所述变体与SEQ ID NO:27所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:37所示的氨基酸序列或其变体，所述变体与SEQ ID NO:37所示的氨基酸序列具有至少约80％序列同一性；(iv)VH，其包含SEQ ID NO:28所示的氨基酸序列或其变体，所述变体与SEQ ID NO:28所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:38所示的氨基酸序列或其变体，所述变体与SEQ ID NO:38所示的氨基酸序列具有至少约80％序列同一性；(v)VH，其包含SEQ ID NO:29所示的氨基酸序列或其变体，所述变体与SEQ ID NO:29所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:39所示的氨基酸序列或其变体，所述变体与SEQ ID NO:39所示的氨基酸序列具有至少约80％序列同一性；(vi)VH，其包含SEQ ID NO:30所示的氨基酸序列或其变体，所述变体与SEQ ID NO:30所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:40所示的氨基酸序列或其变体，所述变体与SEQ ID NO:40所示的氨基酸序列具有至少约80％序列同一性；(vii)VH，其包含SEQ ID NO:31所示的氨基酸序列或其变体，所述变体与SEQ ID NO:31所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:41所示的氨基酸序列或其变体，所述变体与SEQ ID NO:41所示的氨基酸序列具有至少约80％序列同一性；(viii)VH，其包含SEQ ID NO:32所示的氨基酸序列或其变体，所述变体与SEQ ID NO:32所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:42所示的氨基酸序列或其变体，所述变体与SEQ ID NO:42所示的氨基酸序列具有至少约80％序列同一性；(ix)VH，其包含SEQ ID NO:33所示的氨基酸序列或其变体，所述变体与SEQ ID NO:33所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:43所示的氨基酸序列或其变体，所述变体与SEQ ID NO:43所示的氨基酸序列具有至少约80％序列同一性；或(x)VH，其包含SEQ ID NO:34所示的氨基酸序列或其变体，所述变体与SEQ ID NO:34所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:44所示的氨基酸序列或其变体，所述变体与SEQ ID NO:44所示的氨基酸序列具有至少约80％序列同一性。In some embodiments, any of the isolated anti-C5a antibodies as described above, which includes: (i) VH , which includes the amino acid sequence shown in SEQ ID NO: 25 or a variant thereof, which variant is identical to SEQ The amino acid sequence shown in ID NO:25 has at least about 80% sequence identity; and VL , which includes the amino acid sequence shown in SEQ ID NO:35 or a variant thereof, which variant is identical to the amino acid sequence shown in SEQ ID NO:35 The amino acid sequence shown has at least about 80% sequence identity; (ii) V H , which includes the amino acid sequence shown in SEQ ID NO: 26 or a variant thereof, which variant is identical to the amino acid sequence shown in SEQ ID NO: 26 Sequences having at least about 80% sequence identity; and V L comprising the amino acid sequence set forth in SEQ ID NO: 36 or a variant thereof that has at least about 80% identity to the amino acid sequence set forth in SEQ ID NO: 36 % sequence identity; (iii) V H comprising the amino acid sequence set forth in SEQ ID NO: 27 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 27 and VL , which comprises the amino acid sequence shown in SEQ ID NO:37 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:37; (iv) VH , which comprises the amino acid sequence set forth in SEQ ID NO:28 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO:28; and VL , which comprises The amino acid sequence shown in SEQ ID NO: 38 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO: 38; (v) V H comprising SEQ ID NO : The amino acid sequence shown in SEQ ID NO:29 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:29; and VL , which includes the amino acid sequence shown in SEQ ID NO:39 An amino acid sequence or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO: 39; (vi) V H comprising the amino acid sequence shown in SEQ ID NO: 30 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:30; and VL , which includes the amino acid sequence shown in SEQ ID NO:40 or a variant thereof, where The variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO: 40; (vii) V H , which includes the amino acid sequence shown in SEQ ID NO: 31 or a variant thereof, the variant Having at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO:31; and V L comprising the amino acid sequence set forth in SEQ ID NO:41 or a variant thereof that is identical to SEQ ID NO: The amino acid sequence shown in 41 has at least about 80% sequence identity; (viii) V H , which includes the amino acid sequence shown in SEQ ID NO: 32 or a variant thereof, which variant is identical to the amino acid sequence shown in SEQ ID NO: 32 The amino acid sequence has at least about 80% sequence identity; and V L includes the amino acid sequence shown in SEQ ID NO: 42 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO: 42 About 80% sequence identity; (ix) V H comprising the amino acid sequence shown in SEQ ID NO: 33 or a variant thereof that has at least about 80% identity with the amino acid sequence shown in SEQ ID NO: 33 Sequence identity; and V L comprising the amino acid sequence set forth in SEQ ID NO: 43 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 43; or (x) V H comprising the amino acid sequence set forth in SEQ ID NO: 34 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 34; and V L , which comprises the amino acid sequence shown in SEQ ID NO:44 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:44.
在一些实施例中，分离的抗C5a抗体与人C5a的结合的Kd值为0.1pM至1nM。In some embodiments, the isolated anti-C5a antibody binds to human C5a with a Kd value of 0.1 pM to 1 nM.
在一些实施例中，提供一种分离的抗C5a抗体，其与上述任一种分离的抗C5a抗体竞争与C5a的特异性结合。在一些实施例中，提供一种分离的抗C5a抗体，其与上述任一种分离的抗C5a抗体特异性地结合相同的表位。In some embodiments, an isolated anti-C5a antibody is provided that competes with any of the isolated anti-C5a antibodies described above for specific binding to C5a. In some embodiments, an isolated anti-C5a antibody is provided that specifically binds to the same epitope as any of the isolated anti-C5a antibodies described above.
在一些实施例中，如上所述任一种分离的抗C5a抗体，所述分离的抗C5a抗体包含Fc片段。在一些实施例中，所述分离的抗C5a抗体是全长的IgG抗体。在一些实施例中，所述分离的抗C5a抗体是全长的IgG1或IgG4抗体。在一些实施例中，所述分离的抗C5a抗体是嵌合的、全人的或人源化的抗体。在一些实施例中，所述分离的抗C5a抗体是抗原结合片段，所述抗原结合片段选自Fab、Fab’、F(ab)’2、Fab’-SH、单链Fv(scFv)、Fv片段、dAb、Fd、纳米抗体(nanobody)、双链抗体(diabody)和线性抗体组成的组中。In some embodiments, any of the isolated anti-C5a antibodies described above, the isolated anti-C5a antibody comprises an Fc fragment. In some embodiments, the isolated anti-C5a antibody is a full-length IgG antibody. In some embodiments, the isolated anti-C5a antibody is a full-length IgG1 or IgG4 antibody. In some embodiments, the isolated anti-C5a antibody is a chimeric, fully human, or humanized antibody. In some embodiments, the isolated anti-C5a antibody is an antigen-binding fragment selected from the group consisting of Fab, Fab', F(ab)' 2 , Fab'-SH, single chain Fv (scFv), Fv In the group consisting of fragments, dAbs, Fds, nanobodies, diabodies and linear antibodies.
在一些实施例中，提供一种分离的核酸分子，所述核酸分子编码如上所述任一种抗C5a抗体。在一些实施例中，提供一种载体，所述载体包含如上所述任一种核酸分子。在一些实施例中，提供一种宿主细胞，所述宿主细胞包含如上所述任一种抗C5a抗体、如上所述任一种核酸分子或如上所述任一种载体。在一些实施例中，提供一种制备抗C5a抗体的方法，其包含：a)在能有效表达抗C5a抗体的条件下培养上述任一种宿主细胞；和b)从宿主细胞中获得表达的抗C5a抗体。In some embodiments, an isolated nucleic acid molecule encoding any of the anti-C5a antibodies described above is provided. In some embodiments, a vector is provided comprising any of the nucleic acid molecules described above. In some embodiments, a host cell is provided, the host cell comprising any one of the anti-C5a antibodies described above, any one of the nucleic acid molecules described above, or any one of the vectors described above. In some embodiments, a method for preparing an anti-C5a antibody is provided, which includes: a) culturing any of the above host cells under conditions that can effectively express the anti-C5a antibody; and b) obtaining the expressed anti-C5a antibody from the host cell. C5a antibody.
在一些实施例中，提供一种治疗所需个体疾病或病症的方法，包括向所述个体施用有效量的如上所述的任一种抗C5a抗体。在一些实施例中，提供如上所述的任一种抗C5a抗体在制备用于治疗所需个体疾病或病症的药物组合物中的用途。在一些实施例中，提供如上所述的任一种抗C5a抗体或包含抗C5a抗体的药物组合物在制备用于治疗疾病或病症的药物中的用途。在一些实施例中，所述疾病或病症与C5a信号通路有关，包括自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病或病症。在一些实施例中，所述疾病或病症选自例如，炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。In some embodiments, a method of treating a disease or condition in a desired individual is provided, comprising administering to the individual an effective amount of any of the anti-C5a antibodies described above. In some embodiments, provided is the use of any of the anti-C5a antibodies described above in the preparation of a pharmaceutical composition for treating a disease or condition in a desired individual. In some embodiments, the use of any of the anti-C5a antibodies described above or a pharmaceutical composition comprising an anti-C5a antibody in the preparation of a medicament for treating a disease or condition is provided. In some embodiments, the disease or disorder is related to the C5a signaling pathway, including autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplant immune dysregulation diseases or disorders . In some embodiments, the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
在一些实施例中，提供一种偶联物，其包含单克隆抗体以及偶联部分，其中所述单克隆抗体为本文中所述的任一种抗C5a的抗体，所述偶联部分为放射性同位素、荧光物质、发光物质、有色物质或酶。In some embodiments, a conjugate is provided, which includes a monoclonal antibody and a coupling moiety, wherein the monoclonal antibody is any anti-C5a antibody described herein, and the coupling moiety is a radioactive Isotopes, fluorescent substances, luminescent substances, colored substances or enzymes.
同时还提供包含如上所述的任一种抗C5a抗体的药物组合物、试剂盒以及生产制品。Pharmaceutical compositions, kits and manufactured products containing any of the anti-C5a antibodies mentioned above are also provided.
在一些实施例中，试剂盒或制品包含本文中所述的任一种抗C5a抗体或偶联物。In some embodiments, a kit or article of manufacture includes any one of the anti-C5a antibodies or conjugates described herein.
在一些实施例中，涉及本申请中所述的任一种抗C5a抗体或偶联物在制备试剂盒或制品中的用途，所述试剂盒用于检测样品中C5a的存在或其水平。In some embodiments, the use of any of the anti-C5a antibodies or conjugates described in this application is directed to the preparation of a kit or article of manufacture for detecting the presence or level of C5a in a sample.
在一些实施例中，提供一种检测样品中C5a存在或水平的体外方法，所述方法包括：a)在允许抗体和C5a之间形成复合物的条件下将样品与本文中所述的任一种抗C5a抗体或偶联物相接触；b)检测复合物的形成；和c)确定样品中C5a的存在或其水平。In some embodiments, an in vitro method of detecting the presence or level of C5a in a sample is provided, the method comprising: a) contacting the sample with any of the methods described herein under conditions that allow the formation of a complex between the antibody and C5a. contact an anti-C5a antibody or conjugate; b) detect the formation of a complex; and c) determine the presence or level of C5a in the sample.
在一些实施例中，提供一种诊断C5a水平异常的方法，所述方法包括：a)获得待测样品，b)在允许抗体和C5a之间形成复合物的条件下将待测样品与本文中所述的任一种抗C5a抗体或偶联物相接触；c)检测复合物的形成；d)确定待测样品中C5a的水平；和e)判断待测样品中C5a的水平是否高于正常样品中C5a的水平。In some embodiments, a method for diagnosing abnormalities in C5a levels is provided, the method comprising: a) obtaining a sample to be tested, and b) combining the sample to be tested with the method described herein under conditions that allow the formation of a complex between the antibody and C5a. Contact any of the anti-C5a antibodies or conjugates; c) detect the formation of the complex; d) determine the level of C5a in the sample to be tested; and e) determine whether the level of C5a in the sample to be tested is higher than normal Levels of C5a in the sample.
在一些实施例中，所述样品为血清样品或血浆样品。在另外一些实施例中，所述待测样品来源于有需要的患者。在进一步的优选实施例中，所述患者接受过或未接受过C5a抗体治疗。In some embodiments, the sample is a serum sample or plasma sample. In other embodiments, the sample to be tested is derived from a patient in need. In a further preferred embodiment, the patient has or has not received C5a antibody treatment.
附图说明Description of drawings
图1A-1C所示结果为抗C5a抗体RH23、RH25、RH32、RH35、RH36、RH58、RH85、RH86、RH92、RH95与人C5a的结合曲线。The results shown in Figures 1A-1C are the binding curves of anti-C5a antibodies RH23, RH25, RH32, RH35, RH36, RH58, RH85, RH86, RH92, RH95 and human C5a.
图2所示结果为抗C5a抗体RH23、RH25、RH32、RH35、RH36、RH58、RH85、RH86、RH92、RH95与人C5a-desArg的结合曲线。The results shown in Figure 2 are the binding curves of anti-C5a antibodies RH23, RH25, RH32, RH35, RH36, RH58, RH85, RH86, RH92, RH95 and human C5a-desArg.
本申请的详细描述Detailed description of this application
本申请一方面提供抗C5a抗体分子。通过杂交瘤技术、scFv噬菌体库筛选以及适当设计的生物化学及生物学实验的组合，已经鉴定出能够结合人C5a并抑制人C5a与其受体作用的高效抗体分子。本文给出的结果表明，与已知抗C5a抗体INab308相比，本申请中的抗体以高亲和力结合人C5a或人C5a-desArg。In one aspect the application provides anti-C5a antibody molecules. Through a combination of hybridoma technology, scFv phage library screening, and appropriately designed biochemical and biological experiments, highly efficient antibody molecules capable of binding human C5a and inhibiting the interaction of human C5a with its receptor have been identified. The results presented here demonstrate that the antibodies in this application bind human C5a or human C5a-desArg with high affinity compared to the known anti-C5a antibody INab308.
本申请所提供的抗C5a抗体包括，例如，全长抗C5a抗体、抗C5a单链抗体(scFvs)、抗C5a Fc融合蛋白、多特异性(如双特异性)抗C5a抗体、抗C5a免疫偶联物以及诸如此类的。Anti-C5a antibodies provided by this application include, for example, full-length anti-C5a antibodies, anti-C5a single chain antibodies (scFvs), anti-C5a Fc fusion proteins, multispecific (such as bispecific) anti-C5a antibodies, anti-C5a immunoconjugates Connected objects and the like.
另一方面，本申请提供抗C5a抗体，所述抗C5a抗体包含：VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:2，和HC-CDR3，其包含氨基酸序列SEQ ID NO:6，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。On the other hand, the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 2, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 6, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 20, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
另一方面，本申请提供抗C5a抗体，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。On the other hand, the application provides an anti-C5a antibody, the anti-C5a antibody comprising VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L Comprising: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 20, or the V L Variants containing up to about 5 amino acid substitutions in their LC-CDRs.
另一方面，本申请提供抗C5a抗体，所述抗C5a抗体包含：VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:8，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:13，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:21，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。On the other hand, the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 8, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which contains the amino acid sequence SEQ ID NO: 13, LC-CDR2, which contains the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which contains the amino acid sequence SEQ ID NO: 21, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
另一方面，本申请提供抗C5a抗体，所述抗C5a抗体包含：VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。On the other hand, the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 9, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
另一方面，本申请提供抗C5a抗体，所述抗C5a抗体包含：VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:5，和HC-CDR3，其包含氨基酸序列SEQ ID NO:10，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。On the other hand, the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 5, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 10, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
另一方面，本申请提供抗C5a抗体，所述抗C5a抗体包含：VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。On the other hand, the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
另一方面，本申请提供抗C5a抗体，所述抗C5a抗体包含：VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:23，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。On the other hand, the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 9, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 23, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
另一方面，本申请提供抗C5a抗体，所述抗C5a抗体包含：VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:15，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。On the other hand, the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which contains the amino acid sequence SEQ ID NO: 15, LC-CDR2, which contains the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which contains the amino acid sequence SEQ ID NO: 22, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
另一方面，本申请提供抗C5a抗体，所述抗C5a抗体包含：VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。On the other hand, the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
另一方面，本申请提供抗C5a抗体，所述抗C5a抗体包含：VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:11，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:16，LC-CDR2，其包含氨基酸序列SEQ ID NO:19，和LC-CDR3，其包含氨基酸序列SEQ ID NO:24，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。On the other hand, the application provides an anti-C5a antibody, the anti-C5a antibody comprising: VH , the VH comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 11, or a variant of the V H containing up to about 5 amino acid substitutions in its HC-CDRs; and V L , the V L includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 16, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 19, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 24, or the V Variants of L containing up to about 5 amino acid substitutions in their LC-CDRs.
同时还提供编码抗C5a抗体的核酸，包含抗C5a抗体的组合物，以及制备和使用抗C5a抗体的方法。Also provided are nucleic acids encoding anti-C5a antibodies, compositions containing anti-C5a antibodies, and methods of preparing and using anti-C5a antibodies.
定义definition
如本文所述，“治疗(treatment)”或“治疗(treating)是一种获得有益的或期望的结果的方法，包括临床结果。鉴于本申请的目的，所述有益的或期望的临床结果，包括但不限于以下一种或多种：缓解由疾病引起的一种或多种症状，减轻疾病程度，稳定疾病(例如，预防或延迟疾病恶化)，预防或延迟疾病的扩散(例如，转移)，预防或延迟疾病复发，延迟或减缓疾病进展，改善疾病状态，缓解疾病(部分或全部)，减少治疗疾病所需的一种或多种其他药物的剂量，延迟疾病进展，改善或提高生活质量，增加体重，和/或延长生存期。同时，“治疗”还包括疾病病理结果的减少(例如，对癌症而言，肿瘤体积)。本申请的方法考虑了这些治疗的任何一个或多个方面。As used herein, "treatment" or "treating" is a method of obtaining beneficial or desired results, including clinical results. For the purposes of this application, said beneficial or desired clinical results, Including but not limited to one or more of the following: alleviating one or more symptoms caused by the disease, reducing the severity of the disease, stabilizing the disease (e.g., preventing or delaying the progression of the disease), preventing or delaying the spread of the disease (e.g., metastasis) , prevent or delay disease recurrence, delay or slow disease progression, improve disease status, alleviate disease (partial or complete), reduce the dose of one or more other drugs required to treat the disease, delay disease progression, improve or improve quality of life , increase body weight, and/or prolong survival. Also, "treatment" includes reduction of pathological consequences of disease (e.g., for cancer, tumor volume). The methods of the present application contemplate any one or more aspects of these treatments .
术语“抗体”包括全长抗体及其抗原结合片段。全长抗体包括两条重链和两条轻链。轻链和重链的可变区负责抗原的结合。两条链中的可变区通常包括3个高变的环，被称为互补决定区(CDRs)(轻链(LC)CDRs包括LC-CDR1、LC-CDR2和LC-CDR3，重链(HC)CDRs包括HC-CDR1、HC-CDR2和HC-CDR3)。本文所披露的抗体或抗原结合片段的CDR边界可以通过Kabat，Chothia或Al-Lazikani惯例来定义或识别(Al-Lazikani 1997；Chothia 1985；Chothia 1987；Chothia 1989；Kabat 1987；Kabat 1991)。重链或轻链的3个CDR区插入到被称为框架区(FRs)的侧翼区段之间，所述框架区比CDR区具有更高的保守性，并形成支撑高变环的支架。重链和轻链的恒定区并不参与抗原结合，但展示出多种效应功能。抗体是基于它们重链恒定区的氨基酸序列进行分类的。抗体的五种主要类别或同种型是IgA、IgD、IgE、IgG和IgM，其特征在于分别具有α、δ、ε、γ和μ型重链。几种主要的抗体类别被分为亚类，如IgG1(γ1重链)、IgG2(γ2重链)、IgG3(γ3重链)、IgG4(γ4重链)、IgA1(α1重链)或IgA2(α2重链)。The term "antibody" includes full-length antibodies and antigen-binding fragments thereof. Full-length antibodies include two heavy chains and two light chains. The variable regions of the light and heavy chains are responsible for antigen binding. The variable regions in the two chains usually include three hypervariable loops, called complementarity determining regions (CDRs) (light chain (LC) CDRs include LC-CDR1, LC-CDR2 and LC-CDR3, heavy chain (HC) )CDRs include HC-CDR1, HC-CDR2 and HC-CDR3). The CDR boundaries of the antibodies or antigen-binding fragments disclosed herein may be defined or identified by the Kabat, Chothia or Al-Lazikani conventions (Al-Lazikani 1997; Chothia 1985; Chothia 1987; Chothia 1989; Kabat 1987; Kabat 1991). The three CDR regions of the heavy or light chain are inserted between flanking segments called framework regions (FRs), which are more conserved than the CDR regions and form a scaffold supporting the hypervariable loops. The constant regions of the heavy and light chains are not involved in antigen binding but exhibit a variety of effector functions. Antibodies are classified based on the amino acid sequence of their heavy chain constant region. The five major classes or isotypes of antibodies are IgA, IgD, IgE, IgG, and IgM, characterized by heavy chains of the alpha, delta, epsilon, gamma, and mu types, respectively. Several major antibody classes are divided into subclasses, such as IgG1 (γ1 heavy chain), IgG2 (γ2 heavy chain), IgG3 (γ3 heavy chain), IgG4 (γ4 heavy chain), IgA1 (α1 heavy chain), or IgA2 ( α2 heavy chain).
如本文所述，术语“抗原结合片段”包括抗体片段，例如，双链抗体(diabody)、Fab、Fab’、F(ab’)2、Fv片段、二硫键稳定的Fv片段(dsFv)、(dsFv)2、双特异性dsFv(dsFv-dsFv’)、二硫键稳定的双链抗体(ds双链抗体)、单链Fv(scFv)、scFv二聚体(二价双链抗体)，由包含一个或多个CDRs的抗体片段组成的多特异性抗体、单域抗体、纳米抗体(nanobody)、域抗体、二价域抗体或者能够与抗原结合但不包含完整抗体结构的任何其他抗体片段。抗原结合片段能够与亲本抗体或亲本抗体片段(如亲本scFv)结合相同的抗原。抗原结合片段还包括包含上述抗体片段的融合蛋白。在一些实施例中，抗原结合片段可能包括来自特定人抗体的一个或多个CDRs，该CDRs被移植到来自一个或多个不同人抗体的框架区。As used herein, the term "antigen-binding fragment" includes antibody fragments, e.g., diabodies, Fab, Fab', F(ab') 2 , Fv fragments, disulfide-stabilized Fv fragments (dsFv), (dsFv) 2. Bispecific dsFv (dsFv-dsFv'), disulfide bond-stabilized diabody (ds diabody), single-chain Fv (scFv), scFv dimer (bivalent diabody), Multispecific antibodies consisting of antibody fragments containing one or more CDRs, single domain antibodies, nanobodies, domain antibodies, bivalent domain antibodies, or any other antibody fragment capable of binding to an antigen but not containing a complete antibody structure . The antigen-binding fragment is capable of binding to the same antigen as the parent antibody or parent antibody fragment (eg, parent scFv). Antigen-binding fragments also include fusion proteins containing the above-described antibody fragments. In some embodiments, the antigen-binding fragment may include one or more CDRs from a specific human antibody grafted to framework regions from one or more different human antibodies.
如本文所述，术语“表位”是指抗体或抗体部分结合的抗原上特定的原子或氨基酸组。如果两种抗体或抗体部分表现出与某抗原竞争性结合，则它们可能结合抗原上的相同表位。As used herein, the term "epitope" refers to a specific group of atoms or amino acids on an antigen to which an antibody or antibody portion binds. If two antibodies or antibody portions exhibit competitive binding to an antigen, they may bind to the same epitope on the antigen.
如本文所用，当第一抗体在等摩尔浓度下抑制第二抗体与C5a靶标结合至少50％(例如至少55％、60％、65％、70％、75％、80％、85％、90％、95％、98％或99％)时，第一抗体与第二抗体“竞争”结合C5a靶标，反之亦然。PCT出版物WO 03/48731描述了基于交叉竞争的高通量抗体“表位归类”方法。As used herein, a first antibody inhibits binding of a second antibody to a C5a target by at least 50% (e.g., at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%) at equimolar concentrations , 95%, 98% or 99%), the first antibody "competes" with the second antibody for binding to the C5a target, and vice versa. PCT publication WO 03/48731 describes a cross-competition based high-throughput antibody "epitope classification" approach.
如本文所述，术语“特异性地结合”、“特异性地识别”或“对..来说是特异性的”是指可测量的和可再现的相互作用，例如抗体与靶标的结合可以确定在异质分子群，包括生物分子中存在该靶标。例如，抗体能够特异性地识别某靶标(可以是表位)是指，与其它靶标结合相比，该抗体与该靶标的结合具有更高的亲和力，亲合力，更容易和/或更持久。在一些实施例中，特异性地识别抗原的抗体与抗原的一个或多个抗原决定簇反应，其结合亲和力是其与其它靶标结合亲和力的至少10倍。As used herein, the terms "specifically bind," "specifically recognize," or "specific for" refer to a measurable and reproducible interaction, e.g., binding of an antibody to a target can The presence of the target in a heterogeneous population of molecules, including biomolecules, is determined. For example, the ability of an antibody to specifically recognize a target (which may be an epitope) means that the antibody binds to the target with higher affinity, avidity, easier and/or longer duration than to other targets. In some embodiments, an antibody that specifically recognizes an antigen reacts with one or more epitopes of the antigen with a binding affinity that is at least 10 times greater than its binding affinity to other targets.
如本文所述，一种“分离的”抗C5a抗体是指一种抗C5a抗体，其(1)与天然存在的蛋白无关，(2)不含相同来源的其他蛋白，(3)由不同种属的细胞所表达，或(4)自然界中不存在。As used herein, an "isolated" anti-C5a antibody is an anti-C5a antibody that (1) is not related to a naturally occurring protein, (2) does not contain other proteins from the same source, and (3) is produced from a different species Expressed by cells of the genus, or (4) does not exist in nature.
如本文所述，术语“分离的核酸”，是指基因组、cDNA或合成来源的核酸或其组合。根据其来源，所述“分离的核酸”是指(1)与自然界中发现的“分离的核酸”中的全部或部分多核苷酸无关，(2)可与自然状态下不与之相连的多核苷酸可操作性地连接，或(3)在自然界中不作为较长序列的一部分而存在。As used herein, the term "isolated nucleic acid" refers to nucleic acids of genomic, cDNA or synthetic origin, or combinations thereof. Depending on its source, the "isolated nucleic acid" refers to (1) not related to all or part of the polynucleotides in the "isolated nucleic acid" found in nature, (2) may not be related to polynucleotides that are not associated with it in the natural state. The nucleotides are operably linked, or (3) do not occur in nature as part of a longer sequence.
如本文所用，术语“CDR”或“互补决定区”意指重链和轻链多肽的可变结构域内发现的非连续抗原结合位点。在文献Kabat et al.,J.Biol.Chem.252:6609-6616(1977)；Kabat et al.,U.S.Dept.of Health and Human Services,“Sequences ofproteins ofimmunological interest”(1991)；Chothia et al.,J.Mol.Biol.196:901-917(1987)；Al-Lazikani B.et al.,J.Mol.Biol.,273:927-948(1997)；MacCallum et al.,J.Mol.Biol.262:732-745(1996)；Abhinandan and Martin,Mol.Immunol.,45:3832-3839 (2008)；Lefranc M.P.et al.,Dev.Comp.Immunol.,27:55-77(2003)；和Honegger and Plückthun,J.Mol.Biol.,309:657-670(2001)中已经描述这些特殊的区域，其中当彼此之间互相比较时，这些定义包括氨基酸残基的重合或子集。然而，采用任何一种定义方式来指示抗体或移植抗体或其变体的CDR，均包括在本文所定义和使用的术语范围之内。表1中列了由上述引用的各篇参考文献所定义的CDR所包括的氨基酸残基的位置，以示比较。CDR预测的算法和结合界面在本领域是已知的，包括，例如Abhinandan andMartin,Mol.Immunol.,45:3832-3839(2008)；Ehrenmann F.et al.,NucleicAcids Res.,38:D301-D307(2010)；和Adolf-Bryfogle J.et al.,NucleicAcids Res.,43:D432-D438(2015)中均有描述。本段中所引用的参考文献的内容以其整体引用并入本文中，以用于本申请和可能包含在本文中的一个或多个权利要求中。As used herein, the term "CDR" or "complementarity determining region" means the non-contiguous antigen binding sites found within the variable domains of heavy and light chain polypeptides. In the literature Kabat et al., J. Biol. Chem. 252:6609-6616 (1977); Kabat et al., U.S. Dept. of Health and Human Services, "Sequences of proteins of immunological interest" (1991); Chothia et al. ,J.Mol.Biol.196:901-917(1987);Al-Lazikani B.et al.,J.Mol.Biol.,273:927-948(1997);MacCallum et al.,J.Mol. Biol.262:732-745(1996);Abhinandan and Martin, Mol.Immunol.,45:3832-3839 (2008);Lefranc M.P.et al.,Dev.Comp.Immunol.,27:55-77(2003) ; and Honegger and Plückthun, J. Mol. Biol., 309:657-670 (2001), where these definitions include overlapping or subsets of amino acid residues when compared to each other. However, any definition that refers to the CDRs of an antibody or grafted antibody or a variant thereof is included within the scope of the term as defined and used herein. The positions of the amino acid residues included in the CDRs defined by the above-cited references are listed in Table 1 for comparison. Algorithms and binding interfaces for CDR prediction are known in the art, including, for example, Abhinandan and Martin, Mol. Immunol., 45:3832-3839 (2008); Ehrenmann F. et al., Nucleic Acids Res., 38:D301- D307(2010); and Adolf-Bryfogle J.et al., NucleicAcids Res., 43:D432-D438(2015). The contents of the references cited in this paragraph are incorporated by reference in their entirety for purposes of this application and one or more claims that may be included herein.
表1:CDR定义 1氨基酸残基编号参照上述Kabat et al.中的命名方法2氨基酸残基编号参照上述Chothia et al.中的命名方法3氨基酸残基编号参照上述MacCallum et al.中的命名方法4氨基酸残基编号参照上述Lefranc et al.中的命名方法5氨基酸残基编号参照上述Honegger and Plückthun中的命名方法Table 1: CDR definition 1. The numbering of amino acid residues refers to the nomenclature method in Kabat et al. above. 2. The numbering of amino acid residues refers to the nomenclature method in Chothia et al. above. 3. The numbering of amino acid residues refers to the nomenclature method in MacCallum et al. above. 4. The numbering of amino acid residues Refer to the nomenclature method in Lefranc et al. above . 5 Amino acid residue numbers refer to the nomenclature method in Honegger and Plückthun above.
术语“嵌合抗体”是指，重链和/或轻链的一部分与来自特定种属或属于特定抗体种类或亚类的抗体中的相应序列一致或具有同源性，而这个(些)链的剩余部分与来自另一种属或属于其它抗体种类或亚类的抗体中的相应序列一致或具有同源性的抗体，以及此类抗体的片段，只要其具有本申请中的生物学活性(见U.S.PatentNo.4,816,567；andMorrison et al.,Proc.Natl.Acad.Sci.USA,81:6851-6855(1984))。The term "chimeric antibody" means that a portion of the heavy chain and/or light chain is identical or homologous to the corresponding sequence in an antibody from a specific species or belonging to a specific antibody class or subclass, and this chain(s) Antibodies whose remainder are identical to or homologous to corresponding sequences in antibodies from another genus or belonging to other antibody classes or subclasses, as well as fragments of such antibodies, as long as they have the biological activity in this application ( See U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)).
“Fv”是包含完整抗原识别及结合位点的最小抗体片段。该片段是由一个重链可变结构域和一个轻链可变结构域紧密非共价连接形成的二聚体。通过这两个域的折叠衍生出6个高变环(轻链和重链中各3个环)，所述高变环为抗体提供了用于结合抗原的氨基酸残基，并且赋予抗体与抗原结合的特异性。然而，即使单个可变结构域(或Fv片段的一半，其仅包含对抗原具有特异性的3个CDRs)也具有识别和结合抗原的能力，尽管其亲和力低于完整的结合位点。"Fv" is the smallest antibody fragment that contains complete antigen recognition and binding sites. This fragment is a dimer formed by a heavy chain variable domain and a light chain variable domain tightly non-covalently linked. The folding of these two domains results in 6 hypervariable loops (3 loops each in the light chain and heavy chain), which provide the antibody with the amino acid residues for binding to the antigen and confer a binding relationship between the antibody and the antigen. Binding specificity. However, even a single variable domain (or half of an Fv fragment, which contains only the 3 CDRs specific for the antigen) has the ability to recognize and bind the antigen, albeit with lower affinity than the complete binding site.
“单链Fv”，也可简写成“sFv”或“scFv”，是包含被连接成单一多肽链的VH和VL抗体域的抗体片段。在一些实施例中，scFv多肽进一步包括VH和VL域之间的连接多肽，该连接多肽使得scFv形成抗原结合的理想结构。关于scFv的概述，见Pluckthun in The Pharmacology of Monoclonal Antibodies,vol.113,Rosenburg andMoore eds.,Springer-Verlag,New York,pp.269-315(1994)。A "single chain Fv", which may also be abbreviated as "sFv" or "scFv", is an antibody fragment that contains the V H and V L antibody domains linked into a single polypeptide chain. In some embodiments, the scFv polypeptide further includes a linker polypeptide between the VH and VL domains that allows the scFv to form an ideal structure for antigen binding. For an overview of scFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).
术语“双链抗体(diabody)”是在VH和VL域之间采用短接头(例如5～10个残基)，构建scFv片段(见上段内容)制备而成的一种小抗体片段，这样就使得可变结构域在链间而不是链内进行配对，产生一个双价片段，即具有两个抗原结合位点的片段。双特异性的双链抗体是两个“交叉”scFv片段的异二聚体，其中两个抗体的VH和VL域位于不同的多肽链上。在EP 404,097；WO 93/11161；Hollinger et al.,Proc.Natl.Acad.Sci.USA,90:6444-6448(1993)中全面描述了双链抗体。The term "diabody" is a small antibody fragment prepared by using a short linker (for example, 5 to 10 residues) between the V H and V L domains to construct an scFv fragment (see the previous paragraph). This allows the variable domains to pair inter-chain rather than intra-chain, producing a bivalent fragment, that is, a fragment with two antigen-binding sites. Bispecific diabodies are heterodimers of two "cross-over" scFv fragments, in which the VH and VL domains of the two antibodies are located on different polypeptide chains. Diabodies are fully described in EP 404,097; WO 93/11161; Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993).
非人源(如啮齿类)抗体的“人源化”形式是嵌合抗体，其包括最少的来自非人源抗体的序列。大多数情况下，人源化抗体是人源免疫球蛋白(受体抗体)，其中受体抗体的高变区(HVR)残基被来自非人源种属例如小鼠、大鼠、兔或非人类哺乳动物的且具有理想的抗体特异性，亲和力和性能的高变区残基所取代(供体抗体)。在某些情况下，人源免疫球蛋白框架区中的残基被相应的非人源残基所取代。另外，人源化抗体可以包括在受体抗体或供体抗体中均不存在的残基。这些修饰能够进一步改善抗体的性能。通常，人源化抗体会包含基本上所有，至少一个，通常两个可变结构域，其中所有或基本上所有的高变环均与非人免疫球蛋白的高变环相对应，以及所有或基本上所有的框架区均是人免疫球蛋白序列。人源抗体任选地也还包括免疫球蛋白恒定区(Fc)的至少一部分，通常是人免疫球蛋白的恒定区。具体细节可以参考Jones et al.,Nature 321:522-525(1986)；Riechmann et al.,Nature 332:323-329(1988)；和Presta,Curr.Op.Struct.Biol.2:593-596(1992)。"Humanized" forms of non-human (eg, rodent) antibodies are chimeric antibodies, which include minimal sequence from the non-human antibody. In most cases, humanized antibodies are human immunoglobulins (recipient antibodies) in which the hypervariable region (HVR) residues of the receptor antibody are modified from a non-human species such as mouse, rat, rabbit or Replacement of hypervariable region residues that are non-human mammalian and possess the desired antibody specificity, affinity and performance (donor antibody). In some cases, residues in the human immunoglobulin framework region are replaced with corresponding non-human residues. Additionally, humanized antibodies may include residues that are not present in either the recipient antibody or the donor antibody. These modifications can further improve antibody performance. Typically, a humanized antibody will contain substantially all, at least one, and usually two variable domains in which all or substantially all of the hypervariable loops correspond to hypervariable loops of a non-human immunoglobulin, and all or Essentially all framework regions are human immunoglobulin sequences. The human antibody optionally also includes at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For specific details, please refer to Jones et al., Nature 321:522-525(1986); Riechmann et al., Nature 332:323-329(1988); and Presta, Curr.Op.Struct.Biol.2:593-596 (1992).
本文所鉴定的多肽和抗体序列的“氨基酸序列同一性百分比(％)”或“同源性”被定义：在认为保守性取代属于序列同一性的一部分的情况下进行序列对比，候选序列与待比较多肽序列中相同氨基酸残基所占的百分比。可以通过本领域技术范围内的多种比对方式来确定氨基酸序列同一性百分比，例如，使用如BLAST、BLAST-2、ALIGN、Megalign(DNASTAR)、或MUSCLE软件等可公开获得的计算机软件。本领域技术人员可以确定用于测量比对的合适的参数，包括在所比较序列的全长上实现最大化比对所需的任何算法。然而，为了本文的目的，氨基酸序列同一性百分比数值是使用序列比对电脑程序MUSCLE(Edgar,R.C.,Nucleic Acids Research 32(5):1792-1797,2004；Edgar,R.C.,BMC Bioinformatics 5(1):113,2004)生成的。The "percent amino acid sequence identity (%)" or "homology" of the polypeptide and antibody sequences identified herein is defined as follows: Sequence comparisons are made where conservative substitutions are considered part of the sequence identity, and the candidate sequence is compared with the candidate sequence to be Compares the percentage of identical amino acid residues in a polypeptide sequence. Percent amino acid sequence identity can be determined by a variety of alignment methods within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, Megalign (DNASTAR), or MUSCLE software. One skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms required to maximize alignment over the full length of the sequences being compared. However, for the purposes of this article, percent amino acid sequence identity values were determined using the sequence alignment computer program MUSCLE (Edgar, R.C., Nucleic Acids Research 32(5):1792-1797, 2004; Edgar, R.C., BMC Bioinformatics 5(1) :113,2004).
术语“Fc受体”或“FcR”用于描述结合抗体Fc区的受体。在一些实施例中，本申请所述的FcR是结合IgG抗体(一种γ受体)的FcR，包括FcγRI、FcγRII和FcγRIII亚类的受体，包括这些受体的等位基因变体和可变剪接形式。FcγRII受体包括FcγRIIA(“激活受体”)和FcγRIIB(“抑制受体”)，它们具有相似的氨基酸序列，主要在细胞质结构域有所不同。激活受体FcγRIIA的胞质结构域中含有免疫受体酪氨酸活化基序(ITAM)。抑制受体FcγRIIB的胞质结构域中含有免疫受体酪氨酸抑制基序(ITIM)(见M.inAnnu.Rev.Immunol.15:203-234(1997))。所述术语还包括同种异型，例如FcγRIIIA同种异型：FcγRIIIA-Phe158、FcγRIIIA-Val158、FcγRIIA-R131和/或FcγRIIA-H131。在Ravetch and Kinet,Annu.Rev.Immunol 9:457-92(1991)和Capel et al.,Immunomethods 4:25-34(1994)；以及de Haas et al.,J.Lab.Clin.Med.126:330-41(1995)中对FcRs进行了描述。本申请中术语FcR涵盖其他类型的FcRs，包括将来鉴定的FcRs。术语FcR同时还包括新生儿受体FcRn，其负责向新生儿转移母体IgGs(Guyer et al.,J.Immunol.117:587(1976)以及Kim et al.,J.Immunol.24:249(1994))。The term "Fc receptor" or "FcR" is used to describe a receptor that binds to the Fc region of an antibody. In some embodiments, the FcR described herein is an FcR that binds an IgG antibody, a gamma receptor, including receptors of the FcγRI, FcγRII, and FcγRIII subclasses, including allelic variants and agonist variants of these receptors. Change splicing form. FcγRII receptors include FcγRIIA (“activating receptor”) and FcγRIIB (“inhibitory receptor”), which have similar amino acid sequences and differ mainly in the cytoplasmic domain. The cytoplasmic domain of activating receptor FcγRIIA contains an immunoreceptor tyrosine activation motif (ITAM). The cytoplasmic domain of the inhibitory receptor FcγRIIB contains an immunoreceptor tyrosine inhibitory motif (ITIM) (see M.in Annu. Rev. Immunol. 15:203-234 (1997)). The term also includes allotypes, such as FcγRIIIA allotypes: FcγRIIIA-Phel58, FcγRIIIA-Val158, FcγRIIA-R131 and/or FcγRIIA-H131. In Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991) and Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J. Lab. Clin. Med. 126 FcRs are described in :330-41 (1995). The term FcR in this application covers other types of FcRs, including FcRs identified in the future. The term FcR also includes the neonatal receptor FcRn, which is responsible for the transfer of maternal IgGs to the neonate (Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol. 24:249 (1994) )).
术语“FcRn”指新生儿Fc受体(FcRn)。FcRn与主要组织相容性复合体(MHC)在结构上相似，由α链非共价结合到β2微球蛋白上组成。新生儿Fc受体FcRn的多种功能在Ghetie and Ward(2000)Annu.Rev.Immunol.18,739-766.中进行了综述。FcRn在免疫球蛋白IgGs从母体向新生儿的被动转运和调控血清IgG水平中起到重要作用。FcRn作为一种救助受体，可以在细胞内和细胞间以完整的形式结合和运输胞吞化的IgG，并使它们免于经受默认的降解途径。The term "FcRn" refers to the neonatal Fc receptor (FcRn). FcRn is structurally similar to the major histocompatibility complex (MHC) and consists of an α chain non-covalently bound to β2 microglobulin. The diverse functions of the neonatal Fc receptor FcRn are reviewed in Ghetie and Ward (2000) Annu. Rev. Immunol. 18, 739-766. FcRn plays an important role in the passive transport of immunoglobulin IgGs from mother to newborn and in regulating serum IgG levels. FcRn serves as a rescue receptor that binds and transports endocytosed IgG in an intact form within and between cells and protects them from default degradation pathways.
人IgG重链恒定区的“CH1结构域”通常从118位氨基酸延伸到215位氨基酸(EU编号系统)。The "CH1 domain" of the human IgG heavy chain constant region generally extends from amino acid 118 to amino acid 215 (EU numbering system).
“铰链区”通常被定义为从人IgG1的216位Glu延伸到230位Pro(Burton,Molec.Immunol.22:161-206(1985))。通过将形成重链间二硫键的第一个和最后一个半胱氨酸残基置于与IgG1相同位置后，可以使得其他IgG同种型的铰链区与IgG1序列比对。The "hinge region" is generally defined as extending from position Glu 216 to position Pro 230 of human IgG1 (Burton, Molec. Immunol. 22:161-206 (1985)). By placing the first and last cysteine residues that form the inter-heavy chain disulfide bond after the same position as IgG1, the hinge regions of other IgG isotypes can be aligned with the IgG1 sequence.
人IgG Fc区的“CH2结构域”通常从231位氨基酸延伸到340位氨基酸。CH2结构域的独特之处在于，它不会与另一个区域紧密配对，而是在完整的天然IgG分子的两个CH2结构域之间插入了两条N端连接的支链糖链。据推测，糖类可能作为域与域间配对的替代，有助于保持CH2结构域稳定。Burton,Molec.Immunol.22:161-206(1985)。The "CH2 domain" of the human IgG Fc region usually extends from amino acid 231 to amino acid 340. The unique feature of the CH2 domain is that it does not pair closely with another region, but instead has two N-terminal linked branched sugar chains inserted between the two CH2 domains of the intact natural IgG molecule. It is speculated that sugars may serve as a surrogate for domain-to-domain pairing, helping to keep the CH2 domain stable. Burton, Molec. Immunol. 22:161-206 (1985).
“CH3”结构域包括在Fc区内从C末端残基延伸到CH2结构域(从341位氨基酸到抗体序列的C末端，通常为IgG的第446或447位氨基酸残基)。The "CH3" domain includes the region extending from the C-terminal residues within the Fc region to the CH2 domain (from amino acid 341 to the C-terminus of the antibody sequence, usually amino acid residue 446 or 447 of IgG).
“功能性Fc片段”具有天然Fc区序列所具有的“效应功能”。示例性的“效应功能”包括C1q结合；补体依赖的细胞毒作用(CDC)；Fc受体结合；抗体依赖的细胞介导的细胞毒作用(ADCC)；吞噬作用；细胞表面受体的下调(如B细胞受体；BCR)等。这类效应功能通常需要Fc区与结合结构域(如抗体可变区)结合，并且可以使用本领域公知的多种实验方法进行评估。A "functional Fc fragment" has the "effector function" possessed by a native Fc region sequence. Exemplary "effector functions" include Clq binding; complement-dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors ( Such as B cell receptor; BCR), etc. Such effector functions typically require the binding of an Fc region to a binding domain (eg, an antibody variable region) and can be assessed using a variety of experimental methods well known in the art.
具有“改变的”FcR结合亲和力或ADCC活性的IgG Fc变体的抗体，与亲本多肽或包含天然Fc序列的多肽相比，其FcR结合活性和/或ADCC活性增强或减弱。表现出与FcR“结合增强”的Fc变体与亲本多肽或包含天然IgG Fc序列的多肽相比，其与至少一种FcR具有更高的结合亲和力(例如更低的表观Kd或IC50值)。在一些实施例中，与亲本多肽相比，结合能力增强3倍，例如5、10、25、50、60、100、150、200，甚至高达500倍或结合力提高25％到1000％。表现出与FcR“结合降低”的Fc变体，与亲本多肽相比，其与至少一种FcR具有更低的亲和力(例如更高的表观Kd或IC50值)。与亲本多肽相比，其结合能力下降40％或更多。Antibodies with IgG Fc variants that have "altered" FcR binding affinity or ADCC activity have increased or decreased FcR binding activity and/or ADCC activity compared to the parent polypeptide or a polypeptide comprising a native Fc sequence. An Fc variant that exhibits "enhanced binding" to an FcR has a higher binding affinity (e.g., a lower apparent Kd or IC50 value) to at least one FcR compared to the parent polypeptide or a polypeptide comprising a native IgG Fc sequence. ). In some embodiments, the binding capacity is enhanced 3-fold, such as 5, 10, 25, 50, 60, 100, 150, 200, even up to 500-fold or a 25% to 1000% increase in binding capacity compared to the parent polypeptide. An Fc variant exhibits "reduced binding" to an FcR, having a lower affinity (eg, a higher apparent Kd or IC50 value) for at least one FcR compared to the parent polypeptide. Its binding capacity is reduced by 40% or more compared to the parent polypeptide.
“抗体依赖的细胞介导的细胞毒作用”或“ADCC”是一种细胞毒性形式，指分泌型的Ig与存在于某些细胞毒性细胞(例如自然杀伤细胞(NK)、中性粒细胞、和巨噬细胞)上的Fc受体(FcRs)结合，使这些细胞毒性效应细胞能够特异性结合携带抗原的靶细胞，随后使用细胞毒素杀死靶细胞。抗体“武装”细胞毒性细胞并且是这种杀伤所必需的。介导ADCC的主要细胞类型中，NK细胞只表达FcγRIII，而单核细胞表达FcγRI、FcγRII和FcγRIII。在Ravetch and Kinet,Annu.Rev.Immunol 9:457-92(1991)第464页的Table 3中总结了在造血细胞上FcR的表达。评估目标分子的ADCC活性，可以进行体外ADCC实验，在美国专利No.5,500,362或5,821,337中进行了描述。适用于此类实验的效应细胞包括外周血单核细胞(PBMC)和自然杀伤性细胞(NK)。可选地，或者此外，目标分子的ADCC活性也可以在体内进行评估，例如在如Clynes et al.PNAS(USA)95:652-656(1998)中所公开的动物模型中进行了描述。"Antibody-dependent cell-mediated cytotoxicity" or "ADCC" is a form of cytotoxicity that refers to the interaction of secreted Ig cells present on certain cytotoxic cells (e.g., natural killer cells (NK), neutrophils, Binding to Fc receptors (FcRs) on macrophages allows these cytotoxic effector cells to specifically bind to target cells carrying antigens and subsequently kill the target cells using cytotoxins. Antibodies "arm" cytotoxic cells and are required for this killing. Among the main cell types that mediate ADCC, NK cells only express FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. The expression of FcR on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol 9:457-92 (1991). To assess the ADCC activity of a target molecule, an in vitro ADCC assay can be performed, as described in U.S. Patent No. 5,500,362 or 5,821,337. Effector cells suitable for such experiments include peripheral blood mononuclear cells (PBMC) and natural killer cells (NK). Alternatively, or in addition, the ADCC activity of the target molecule can also be assessed in vivo, for example in an animal model as disclosed in Clynes et al. PNAS (USA) 95:652-656 (1998).
包含Fc区变体的多肽与包含野生型IgG Fc多肽或亲本多肽相比，在人体效应细胞存在下表现出“增强的ADCC活性”或能够更有效的介导ADCC效应，所述包含Fc区变体的多肽在实验时与包含野生型IgG Fc多肽(或亲本多肽)数量上基本相同时，无论在体外或体内均能更有效的介导ADCC。通常采用本领域已知的任何体外ADCC实验方法来鉴定此类变体，例如用于鉴定ADCC活性的实验或方法，例如在动物模型中等。在一些实施例中，此类变体与野生型Fc(或亲代多肽)相比，介导ADCC的效率提高5到100倍，例如25到50倍。A polypeptide comprising an Fc region variant that exhibits "enhanced ADCC activity" or is able to mediate ADCC effects more efficiently in the presence of human effector cells than a wild-type IgG Fc polypeptide or a parent polypeptide that contains an Fc region variant When the antibody polypeptide is essentially the same in quantity as the wild-type IgG Fc polypeptide (or parent polypeptide) during the experiment, it can more effectively mediate ADCC both in vitro and in vivo. Such variants are generally identified using any in vitro ADCC assay known in the art, such as assays or methods for identifying ADCC activity, such as in animal models and the like. In some embodiments, such variants mediate ADCC 5- to 100-fold, for example, 25- to 50-fold more efficiently than wild-type Fc (or the parent polypeptide).
“补体依赖的细胞毒作用”或“CDC”是指在补体存在的情况下裂解靶细胞。经典的补体途径的激活是由补体系统第一组分(C1q)与结合同源抗原的抗体(具有适宜结构的亚类)相结合而启动的。为了评估补体激活，可以进行CDC实验，如Gazzano-Santoro et al.,J.Immunol.Methods 202:163(1996)中所描述的。在美国专利No.6,194,551B1和WO99/51642中描述了具有改变的Fc区氨基酸序列并增加或降低的C1q结合能力的多肽变体。这些专利出版物的内容通过引用明确地并入本文中。另见Idusogie et al.J.Immunol.164:4178-4184(2000)。"Complement-dependent cytotoxicity" or "CDC" refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway is initiated by the binding of the first component of the complement system (C1q) to antibodies (subclasses with appropriate structures) that bind cognate antigens. To assess complement activation, CDC experiments can be performed as described in Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996). Polypeptide variants with altered Fc region amino acid sequences and increased or decreased Clq binding capacity are described in US Patent No. 6,194,551 Bl and WO99/51642. The contents of these patent publications are expressly incorporated herein by reference. See also Idusogie et al. J. Immunol. 164:4178-4184 (2000).
除非另有说明，一种“编码氨基酸序列的核苷酸序列”包括相互之间互为简并形式且编码相同氨基酸序列的所有核苷酸序列。编码蛋白质或RNA的核苷酸序列也可包括内含子，例如编码蛋白质的核苷酸序列在某些形式中包含内含子。Unless otherwise stated, a "nucleotide sequence encoding an amino acid sequence" includes all nucleotide sequences that are degenerates of each other and encode the same amino acid sequence. Nucleotide sequences encoding proteins or RNA may also include introns, for example, nucleotide sequences encoding proteins may include introns in some forms.
术语“可操作性地连接”是指调控序列与异源核苷酸序列之间的功能性连接，从而使后者表达。例如，当第一个核苷酸序列与第二个核苷酸序列处于功能性关系时，第一个核苷酸序列与第二个核苷酸序列为可操作性地连接。例如，如果启动子影响编码序列的转录或表达，该启动子与编码序列为可操作性地连接。通常，可操作性连接的DNA序列是连续的，并且在必要时，可以在同一个阅读框中连接两个蛋白质编码区。The term "operably linked" refers to a functional linkage between a regulatory sequence and a heterologous nucleotide sequence, thereby allowing expression of the latter. For example, a first nucleotide sequence is operably linked to a second nucleotide sequence when the first nucleotide sequence is in a functional relationship with the second nucleotide sequence. For example, a promoter is operably linked to a coding sequence if it affects the transcription or expression of the coding sequence. Typically, operably linked DNA sequences are contiguous and, if necessary, two protein-coding regions can be joined in the same reading frame.
“同源”是指两个多肽之间或两个核酸分子之间的序列相似性或序列同一性。如果两个比较序列的同一位置为相同的碱基或氨基酸单体亚基时，例如两个DNA分子的同一位置均为腺嘌呤，则这两个DNA分子在该位置是同源的。两个序列间的同源百分比是指两个序列中共有的匹配或同源位置的数量与位置总数之比再乘以100所得函数。例如，两个序列中如果10个位置中有6个位置是相匹配或同源的，则这两个序列的同源性为60％。举例来说，DNA序列ATTGCC和TATGGC具有50％的同源性。通常来说，在比对两个序列时，以得到最大同源性为目的来进行对比。"Homology" refers to sequence similarity or sequence identity between two polypeptides or between two nucleic acid molecules. If the same position of two compared sequences contains the same base or amino acid monomer subunit, for example, the same position of two DNA molecules both contains adenine, then the two DNA molecules are homologous at that position. The percent homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the total number of positions multiplied by 100. For example, if 6 out of 10 positions in two sequences match or are homologous, the two sequences are 60% homologous. For example, the DNA sequences ATTGCC and TATGGC have 50% homology. Generally speaking, when comparing two sequences, the comparison is performed with the purpose of obtaining maximum homology.
本文所公开的抗C5a抗体或组合物的“有效量”是指足以实现特定目的的量。“有效量”可以凭经验和通过已知的与所述目的相关的方法确定。An "effective amount" of an anti-C5a antibody or composition disclosed herein is an amount sufficient to achieve a specified purpose. An "effective amount" can be determined empirically and by methods known to be relevant to the stated purpose.
术语“治疗有效量”是指本文所公开的抗C5a抗体或其组合物能够有效治疗个体的疾病或者症状的用量。例如在癌症的情况中，抗C5a抗体或其组合物的治疗有效量是指能够减少癌细胞数量；减小肿瘤的大小或重量；抑制(即在一定程度上减缓或优选停止)肿瘤细胞对周边器官的浸润；抑制(即在一定程度上减缓或优选停止)肿瘤转移；在一定程度上抑制肿瘤的生长，和/或在一定程度上缓解与癌症相关的一种或多种症状。本文所公开的抗C5a抗体或其组合物在某种程度上能够阻止和/或杀死现有的肿瘤细胞，它可以是细胞抑制性的或细胞毒性的。在一些实施例中，治疗有效量是指能够延长患者生存期的用量。在一些实施例中，治疗有效量是指能够改善患者无进展生存期的用量。The term "therapeutically effective amount" refers to an amount of an anti-C5a antibody or composition thereof disclosed herein that is effective in treating a disease or condition in an individual. For example, in the context of cancer, a therapeutically effective amount of an anti-C5a antibody or composition thereof is one that reduces the number of cancer cells; reduces the size or weight of the tumor; inhibits (i.e., slows or preferably stops to a certain extent) the impact of tumor cells on surrounding cells. Invasion of organs; inhibition (i.e., slowing down or preferably stopping to a certain extent) tumor metastasis; inhibiting tumor growth to a certain extent, and/or alleviating one or more symptoms associated with cancer to a certain extent. The anti-C5a antibodies or compositions thereof disclosed herein are capable of preventing and/or killing existing tumor cells to the extent that they may be cytostatic or cytotoxic. In some embodiments, a therapeutically effective amount refers to an amount capable of prolonging the survival of a patient. In some embodiments, a therapeutically effective amount refers to an amount capable of improving progression-free survival of a patient.
如本文所用的，“药学上可接受的”或“药理学上相容的”是指无生物学活性或者其它不期望性质的材料，例如该材料能够加入到给予患者的药物组合物中，而不会引起显著的不良生物反应，或者，不与组合物中包含的任何其它组分以有害的方式相互作用。药学上可接受的载体或赋形剂优选满足毒理学或制造检测的所需标准和/或包含在美国食品和药品管理局编制的非活性成分指南中。As used herein, "pharmaceutically acceptable" or "pharmacologically compatible" refers to a material that has no biological activity or other undesirable properties, e.g., the material can be incorporated into a pharmaceutical composition administered to a patient, and Does not cause significant adverse biological reactions or interact in a deleterious manner with any other components contained in the composition. Pharmaceutically acceptable carriers or excipients preferably meet required standards for toxicological or manufacturing testing and/or are included in the Inactive Ingredient Guidelines prepared by the U.S. Food and Drug Administration.
本文中描述的本申请的实施例应理解为包含“由……组成”和/或“基本上由……组成”的实施例。Embodiments of the application described herein should be understood to include embodiments "consisting of" and/or "consisting essentially of."
本文中提及“约”为一个数值或参数，包含(和描述)针对该值或参数本身的变体。例如，涉及“约X”的描述，包括“X”的描述。References herein to "about" a value or parameter include (and describe) variations on the value or parameter itself. For example, descriptions referring to "about X" include descriptions of "X".
如本文所用的，提及“不是(not)”一个数值或参数，通常表示并描述“除了(other than)”某一数值或参数之外。例如，该方法不能用于治疗X型癌症，意味着该方法通常用于治疗除X型癌症之外的其他类型的癌症。As used herein, reference to "not" a value or parameter generally means and describes "other than" a value or parameter. For example, the method cannot be used to treat type X cancer, meaning the method is typically used to treat other types of cancer besides type X cancer.
除非上下文另有明确说明，本文和所述权利要求中所采用的单数形式“一”，“一个”和“该”包括复数对象。As used herein and in the claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.
抗C5a抗体anti-C5a antibody
一方面，本申请提供特异性结合C5a的抗C5a抗体。所述抗C5a抗体包括，但不限于，人源化抗体，嵌合抗体，小鼠抗体，人抗体，以及本文所述的包含重链和/或轻链CDRs的抗体分子。一方面，本申请提供与C5a结合的分离的抗体。预期的抗C5a抗体包括，例如，全长抗C5a抗体(如全长IgG1或IgG4)，抗C5a单链抗体，抗C5a Fc融合蛋白，多特异性(如双特异性)抗C5a抗体，抗C5a免疫偶联物，以及诸如此类的。在一些实施例中，抗C5a抗体是全长抗体(如全长IgG1或IgG4)或其抗原结合片段，其特异性结合C5a。在一些实施例中，抗C5a抗体是Fab、Fab’、F(ab)’2、Fab’-SH、单链Fv(scFv)、Fv片段、dAb、Fd、纳米抗体(nanobody)、双链抗体(diabody)或线性抗体。在一些实施例中，特异性结合C5a的抗体是指抗体与C5a结合的亲和力至少是与非靶标结合亲和力的10倍以上(包括例如10、102、103、104、105、106、或107倍)。在一些实施例中，非靶标是指不是C5a的抗原。结合亲和力可通过本领域已知的方法来测定，如ELISA，荧光激活细胞分选(FACS)分析或放射免疫沉淀分析(RIA)。Kd值可以通过本领域已知的方法来测定，如表面等离子共振(SPR)技术或生物层干涉技术(BLI)。In one aspect, the application provides anti-C5a antibodies that specifically bind C5a. Such anti-C5a antibodies include, but are not limited to, humanized antibodies, chimeric antibodies, mouse antibodies, human antibodies, and antibody molecules comprising heavy chain and/or light chain CDRs as described herein. In one aspect, the application provides isolated antibodies that bind C5a. Contemplated anti-C5a antibodies include, for example, full-length anti-C5a antibodies (e.g., full-length IgG1 or IgG4), anti-C5a single chain antibodies, anti-C5a Fc fusion proteins, multispecific (e.g., bispecific) anti-C5a antibodies, anti-C5a Immunoconjugates, and the like. In some embodiments, the anti-C5a antibody is a full-length antibody (eg, full-length IgG1 or IgG4) or an antigen-binding fragment thereof that specifically binds C5a. In some embodiments, the anti-C5a antibody is Fab, Fab', F(ab)' 2 , Fab'-SH, single chain Fv (scFv), Fv fragment, dAb, Fd, nanobody, diabody (diabody) or linear antibody. In some embodiments, an antibody that specifically binds to C5a means that the affinity of the antibody for binding to C5a is at least 10 times greater than the binding affinity for the non-target (including, for example, 10, 10 2 , 10 3 , 10 4 , 10 5 , 10 6 , or 10 7 times). In some embodiments, non-target refers to an antigen that is not C5a. Binding affinity can be determined by methods known in the art, such as ELISA, fluorescence-activated cell sorting (FACS) analysis or radioimmunoprecipitation analysis (RIA). The Kd value can be determined by methods known in the art, such as surface plasmon resonance (SPR) technology or biolayer interference (BLI) technology.
尽管本文广泛地讨论了包含人序列的抗C5a抗体(例如，包含人CDR序列的人重链和轻链可变结构域)，但同时也考虑了非人抗C5a抗体。在一些实施例中，非人抗C5a抗体包括本文所述的抗C5a抗体的人CDR序列和非人框架区序列，在一些实施例中，非人框架区序列包括任何的用于使用如本文所述的一种或多种人CDR序列产生重链和/或轻链可变结构域的序列，包括例如哺乳动物，例如小鼠、大鼠、兔子、猪、牛(例如，牛、公牛、水牛)、鹿、绵羊、山羊、鸡、猫、狗、雪貂、灵长类(例如，小猿，猕猴)等。在一些实施例中，非人抗C5a抗体包括将一种或多种本文所述的人CDR序列移植到非人框架区中(例如，鼠或鸡的框架区序列)所产生的抗C5a抗体。Although anti-C5a antibodies comprising human sequences are discussed broadly herein (eg, human heavy and light chain variable domains comprising human CDR sequences), non-human anti-C5a antibodies are also contemplated. In some embodiments, non-human anti-C5a antibodies include the human CDR sequences and non-human framework sequences of the anti-C5a antibodies described herein. In some embodiments, the non-human framework sequences include any for use as described herein. One or more of the human CDR sequences described above generate sequences of heavy and/or light chain variable domains, including, for example, mammals, such as mice, rats, rabbits, pigs, cattle (e.g., cattle, bulls, buffalo ), deer, sheep, goats, chickens, cats, dogs, ferrets, primates (e.g., small apes, macaques), etc. In some embodiments, non-human anti-C5a antibodies include anti-C5a antibodies generated by grafting one or more human CDR sequences described herein into non-human framework regions (eg, murine or chicken framework region sequences).
示例性人C5a的完整氨基酸序列包含SEQ ID NO：49所示的氨基酸序列或由SEQ ID NO：49所示的氨基酸序列组成。The complete amino acid sequence of an exemplary human C5a includes or consists of the amino acid sequence shown in SEQ ID NO: 49.
在一些实施例中，本文所述抗C5a抗体特异性识别人C5a中的表位。在一些实施例中，所述抗C5a抗体与除人之外其它物种的C5a发生交叉反应。在一些实施例中，所述抗C5a抗体对人C5a是完全特异性的，并且不与其它非人物种发生交叉反应。In some embodiments, the anti-C5a antibodies described herein specifically recognize an epitope in human C5a. In some embodiments, the anti-C5a antibody cross-reacts with C5a of species other than human. In some embodiments, the anti-C5a antibody is fully specific for human C5a and does not cross-react with other non-human species.
在一些实施例中，所述抗C5a抗体与C5a蛋白(或其片段)的至少一种等位基因变体交叉反应。在一些实施例中，等位基因变体与天然存在的C5a蛋白(或其片段)相比，具有至多30个(如1、2、3、4、5、6、7、8、9、10、15、20、25或30个)的氨基酸取代(例如保守取代)。在一些实施例中，所述抗C5a抗体不与C5a蛋白(或其片段)的任何等位基因变体发生交叉反应。In some embodiments, the anti-C5a antibody cross-reacts with at least one allelic variant of the C5a protein (or fragment thereof). In some embodiments, the allelic variant has up to 30 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) compared to the naturally occurring C5a protein (or fragment thereof). , 15, 20, 25 or 30) amino acid substitutions (eg, conservative substitutions). In some embodiments, the anti-C5a antibody does not cross-react with any allelic variant of the C5a protein (or fragment thereof).
在一些实施例中，所述抗C5a抗体与C5a蛋白的至少一种种间变体发生交叉反应。在一些实施例中，例如，C5a蛋白(或其片段)是人C5a，并且C5a蛋白(或其片段)的种间变体是食蟹猴中的变体。在一些实施例中，所述抗C5a抗体不与C5a蛋白的任何种间变体发生交叉反应。In some embodiments, the anti-C5a antibody cross-reacts with at least one interspecies variant of the C5a protein. In some embodiments, for example, the C5a protein (or fragment thereof) is human C5a, and the interspecies variant of the C5a protein (or fragment thereof) is a variant in cynomolgus monkey. In some embodiments, the anti-C5a antibody does not cross-react with any interspecies variant of the C5a protein.
在一些实施例中，如本文所述的任一抗C5a抗体，所述抗C5a抗体包含抗体重链恒定区和抗体轻链恒定区。在一些实施例中，所述抗C5a抗体包含IgG1型重链恒定区。在一些实施例中，所述抗C5a抗体包含IgG2型重链恒定区。在一些实施例中，所述抗C5a抗体包含IgG3型重链恒定区。在一些实施例中，所述抗C5a抗体包含IgG4型重链恒定区。在一些实施例中，所述重链恒定区包含(包括由…组成或基本上由…组成)氨基酸序列SEQ ID NO:45。在一些实施例中，所述重链恒定区包含(包括由…组成或基本上由…组成)氨基酸序列SEQ ID NO:46。在一些实施例中，所述抗C5a抗体包含κ轻链恒定区。在一些实施例中，所述轻链恒定区包含(包括由…组成或基本上由…组成)氨基酸序列SEQ ID NO:47。在一些实施例中，所述抗C5a抗体包含λ轻链恒定区。在一些实施例中，所述轻链恒定区包含(包括由…组成或基本上由…组成)氨基酸序列SEQ ID NO:48。在一些实施例中，所述抗C5a抗体包含抗体重链可变结构域和抗体轻链可变结构域。In some embodiments, as any anti-C5a antibody described herein, the anti-C5a antibody comprises an antibody heavy chain constant region and an antibody light chain constant region. In some embodiments, the anti-C5a antibody comprises an IgGl type heavy chain constant region. In some embodiments, the anti-C5a antibody comprises an IgG2-type heavy chain constant region. In some embodiments, the anti-C5a antibody comprises an IgG3 heavy chain constant region. In some embodiments, the anti-C5a antibody comprises an IgG4 type heavy chain constant region. In some embodiments, the heavy chain constant region comprises (including consists of or consists essentially of) the amino acid sequence SEQ ID NO: 45. In some embodiments, the heavy chain constant region comprises (including consists of or consists essentially of) the amino acid sequence SEQ ID NO: 46. In some embodiments, the anti-C5a antibody comprises a kappa light chain constant region. In some embodiments, the light chain constant region comprises (including consists of or consists essentially of) the amino acid sequence SEQ ID NO: 47. In some embodiments, the anti-C5a antibody comprises a lambda light chain constant region. In some embodiments, the light chain constant region comprises (including consists of or consists essentially of) the amino acid sequence SEQ ID NO: 48. In some embodiments, the anti-C5a antibody comprises an antibody heavy chain variable domain and an antibody light chain variable domain.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:2，HC-CDR3，其包含氨基酸序列SEQ ID NO:6，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，LC-CDR3，其包含氨基酸序列SEQ ID NO:20，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 6, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 20, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:2，和HC-CDR3，其包含氨基酸序列SEQ ID NO:6；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20。In some embodiments, the anti-C5a antibody comprises a VH comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 2, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:6; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:20.
在一些实施例中，所述抗C5a抗体包含VH，其包含如氨基酸序列SEQ ID NO:25所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:35所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3。In some embodiments, the anti-C5a antibody comprises a V H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in the amino acid sequence of SEQ ID NO: 25; and a V L comprising VL as shown in the amino acid sequence SEQ ID NO:35 includes LC-CDR1, LC-CDR2 and LC-CDR3.
在一些实施例中，所述抗C5a抗体包含：VH，所述VH包含氨基酸序列SEQ ID NO:25或其变体，所述变体与氨基酸序列SEQ ID NO:25具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及VL，所述VL包含氨基酸序列SEQ ID NO:35或其变体，所述变体与氨基酸序列SEQ ID NO:35具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述抗C5a抗体包含VH，所述VH包含氨基酸序列SEQ ID NO:25，以及VL，所述VL包含氨基酸序列SEQ ID NO:35。 In some embodiments, the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 25 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 25 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 35 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 35. In some embodiments, the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:25, and a VL comprising the amino acid sequence SEQ ID NO:35.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，LC-CDR3，其包含氨基酸序列SEQ ID NO:20，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, HC -CDR3, which comprises the amino acid sequence SEQ ID NO:7, or a variant of said VH , whose HC-CDRs comprise up to about 5 amino acid substitutions; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 20, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20。In some embodiments, the anti-C5a antibody comprises a VH comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:7; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:20.
在一些实施例中，所述抗C5a抗体包含VH，其包含如氨基酸序列SEQ ID NO:26所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:36所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3。In some embodiments, the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 as shown in the amino acid sequence of SEQ ID NO:26; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO:36 includes LC-CDR1, LC-CDR2 and LC-CDR3.
在一些实施例中，所述抗C5a抗体包含：VH，所述VH包含氨基酸序列SEQ ID NO:26或其变体，所述变体与氨基酸序列SEQ ID NO:26具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及VL，所述VL包含氨基酸序列SEQ ID NO:36或其变体，所述变体与氨基酸序列SEQ ID NO:36具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述抗C5a抗体包含VH，所述VH包含氨基酸序列SEQ ID NO:26，以及VL，所述VL包含氨基酸序列SEQ ID NO:36。 In some embodiments, the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 26 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 26 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 36 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 36. In some embodiments, the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:26, and a VL comprising the amino acid sequence SEQ ID NO:36.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，HC-CDR3，其包含氨基酸序列SEQ ID NO:8，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:13，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，LC-CDR3，其包含氨基酸序列SEQ ID NO:21，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 8, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 13, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 21, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:8；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:13，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:21。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:8; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:13, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:21.
在一些实施例中，所述抗C5a抗体包含VH，其包含如氨基酸序列SEQ ID NO:27所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:37所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3。In some embodiments, the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 as shown in the amino acid sequence of SEQ ID NO:27; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO:37 includes LC-CDR1, LC-CDR2 and LC-CDR3.
在一些实施例中，所述抗C5a抗体包含：VH，所述VH包含氨基酸序列SEQ ID NO:27或其变体，所述变体与氨基酸序列SEQ ID NO:27具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及VL，所述VL包含氨基酸序列SEQ ID NO:37或其变体，所述变体与氨基酸序列SEQ ID NO:37具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述抗C5a抗体包含VH，所述VH包含氨基酸序列SEQ ID NO:27，以及VL，所述VL包含氨基酸序列SEQ ID NO:37。 In some embodiments, the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 27 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 27 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 37 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 37. In some embodiments, the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:27, and a VL comprising the amino acid sequence SEQ ID NO:37.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，HC-CDR3，其包含氨基酸序列SEQ ID NO:9，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 9, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:9; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:22.
在一些实施例中，所述抗C5a抗体包含VH，其包含如氨基酸序列SEQ ID NO:28所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:38所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3。In some embodiments, the anti-C5a antibody comprises a V H comprising HC-CDR1, HC-CDR2 and HC-CDR3 as shown in the amino acid sequence of SEQ ID NO : 28; and a V L comprising VL as shown in the amino acid sequence SEQ ID NO:38 contains LC-CDR1, LC-CDR2 and LC-CDR3.
在一些实施例中，所述抗C5a抗体包含：VH，所述VH包含氨基酸序列SEQ ID NO:28或其变体，所述变体与氨基酸序列SEQ ID NO:28具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及VL，所述VL包含氨基酸序列SEQ ID NO:38或其变体，所述变体与氨基酸序列SEQ ID NO:38具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述抗C5a抗体包含VH，所述VH包含氨基酸序列SEQ ID NO:28，以及VL，所述VL包含氨基酸序列SEQ ID NO:38。 In some embodiments, the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 28 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 28 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 38 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 38. In some embodiments, the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:28, and a VL comprising the amino acid sequence SEQ ID NO:38.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:5，HC-CDR3，其包含氨基酸序列SEQ ID NO:10，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 5, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 10, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:5，和HC-CDR3，其包含氨基酸序列SEQ ID NO:10；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 5, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 10; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO: 14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:22.
在一些实施例中，所述抗C5a抗体包含VH，其包含如氨基酸序列SEQ ID NO:29所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:39所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3。In some embodiments, the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 as shown in the amino acid sequence of SEQ ID NO:29; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO:39 includes LC-CDR1, LC-CDR2 and LC-CDR3.
在一些实施例中，所述抗C5a抗体包含：VH，所述VH包含氨基酸序列SEQ ID NO:29或其变体，所述变体与氨基酸序列SEQ ID NO:29具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及VL，所述VL包含氨基酸序列SEQ ID NO:39或其变体，所述变体与氨基酸序列SEQ ID NO:39具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述抗C5a抗体包含VH，所述VH包含氨基酸序列SEQ ID NO:29，以及VL，所述VL包含氨基酸序列SEQ ID NO:39。 In some embodiments, the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 29 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 29 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 39 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 39. In some embodiments, the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:29, and a VL comprising the amino acid sequence SEQ ID NO:39.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。In some embodiments, the anti-C5a antibody comprises a VH comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:7; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:22.
在一些实施例中，所述抗C5a抗体包含VH，其包含如氨基酸序列SEQ ID NO:30所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:40所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3。In some embodiments, the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 as shown in the amino acid sequence of SEQ ID NO:30; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO:40 includes LC-CDR1, LC-CDR2 and LC-CDR3.
在一些实施例中，所述抗C5a抗体包含：VH，所述VH包含氨基酸序列SEQ ID NO:30或其变体，所述变体与氨基酸序列SEQ ID NO:30具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及VL，所述VL包含氨基酸序列SEQ ID NO:40或其变体，所述变体与氨基酸序列SEQ ID NO:40具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述抗C5a抗体包含VH，所述VH包含氨基酸序列SEQ ID NO:30，以及VL，所述VL包含氨基酸序列SEQ ID NO:40。In some embodiments, the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 30 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 30 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 40 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 40. In some embodiments, the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:30, and a VL comprising the amino acid sequence SEQ ID NO:40.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，HC-CDR3，其包含氨基酸序列SEQ ID NO:9，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，LC-CDR3，其包含氨基酸序列SEQ ID NO:23，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 9, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 23, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:23。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:9; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:12, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:23.
在一些实施例中，所述抗C5a抗体包含VH，其包含如氨基酸序列SEQ ID NO:31所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:41所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3。In some embodiments, the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in the amino acid sequence SEQ ID NO:31 of VH ; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO: 41 includes LC-CDR1, LC-CDR2 and LC-CDR3.
在一些实施例中，所述抗C5a抗体包含：VH，所述VH包含氨基酸序列SEQ ID NO:31或其变体，所述变体与氨基酸序列SEQ ID NO:31具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及VL，所述VL包含氨基酸序列SEQ ID NO:41或其变体，所述变体与氨基酸序列SEQ ID NO:41具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述抗C5a抗体包含VH，所述VH包含氨基酸序列SEQ ID NO:31，以及VL，所述VL包含氨基酸序列SEQ ID NO:41。 In some embodiments, the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 31 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 31 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 41 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 41. In some embodiments, the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:31, and a VL comprising the amino acid sequence SEQ ID NO:41.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:15，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, HC -CDR3, which comprises the amino acid sequence SEQ ID NO:7, or a variant of said VH , whose HC-CDRs comprise up to about 5 amino acid substitutions; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 15, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:15，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。In some embodiments, the anti-C5a antibody comprises a VH comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:7; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:15, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 17, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:22.
在一些实施例中，所述抗C5a抗体包含VH，其包含如氨基酸序列SEQ ID NO:32所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:42所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3。In some embodiments, the anti-C5a antibody comprises a V H comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in the amino acid sequence of SEQ ID NO: 32; and a V L comprising VL as shown in the amino acid sequence SEQ ID NO: 42 includes LC-CDR1, LC-CDR2 and LC-CDR3.
在一些实施例中，所述抗C5a抗体包含：VH，所述VH包含氨基酸序列SEQ ID NO:32或其变体，所述变体与氨基酸序列SEQ ID NO:32具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及VL，所述VL包含氨基酸序列SEQ ID NO:42或其变体，所述变体与氨基酸序列SEQ ID NO:42具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述抗C5a抗体包含VH，所述VH包含氨基酸序列SEQ ID NO:32，以及VL，所述VL包含氨基酸序列SEQ ID NO:42。 In some embodiments, the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 32 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 32 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and V L comprising the amino acid sequence SEQ ID NO: 42 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 42. In some embodiments, the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:32, and a VL comprising the amino acid sequence SEQ ID NO:42.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, HC -CDR3, which comprises the amino acid sequence SEQ ID NO:7, or a variant of said VH , whose HC-CDRs comprise up to about 5 amino acid substitutions; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO:7; and VL , which VL comprises: LC-CDR1, which comprises the amino acid sequence SEQ ID NO:14, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:22.
在一些实施例中，所述抗C5a抗体包含VH，其包含如氨基酸序列SEQ ID NO:33所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:43所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3。In some embodiments, the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in the amino acid sequence SEQ ID NO:33 of VH ; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO: 43 includes LC-CDR1, LC-CDR2 and LC-CDR3.
在一些实施例中，所述抗C5a抗体包含：VH，所述VH包含氨基酸序列SEQ ID NO:33或其变体，所述变体与氨基酸序列SEQ ID NO:33具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及VL，所述VL包含氨基酸序列SEQ ID NO:43或其变体，所述变体与氨基酸序列SEQ ID NO:43具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述抗C5a抗体包含VH，所述VH包含氨基酸序列SEQ ID NO:33，以及VL，所述VL包含氨基酸序列SEQ ID NO:43。 In some embodiments, the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 33 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 33 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 43 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 43. In some embodiments, the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:33, and a VL comprising the amino acid sequence SEQ ID NO:43.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，HC-CDR3，其包含氨基酸序列SEQ ID NO:11，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:16，LC-CDR2，其包含氨基酸序列SEQ ID NO:19，LC-CDR3，其包含氨基酸序列SEQ ID NO:24，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。In some embodiments, the anti-C5a antibody comprises a V H comprising: HC- CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, HC -CDR3, which comprises the amino acid sequence SEQ ID NO: 11, or a variant of said VH , comprising up to about 5 amino acid substitutions in its HC-CDRs; and VL , said VL comprising: LC-CDR1, It comprises the amino acid sequence SEQ ID NO: 16, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 19, LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 24, or a variant of said V L , whose LC- The CDRs contain substitutions of up to about 5 amino acids.
在一些实施例中，所述抗C5a抗体包含VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:11；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:16，LC-CDR2，其包含氨基酸序列SEQ ID NO:19，和LC-CDR3，其包含氨基酸序列SEQ ID NO:24。In some embodiments, the anti-C5a antibody comprises a VH comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 11; and VL, which VL comprises : LC-CDR1, which comprises the amino acid sequence SEQ ID NO: 16, LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 19, and LC-CDR3 comprising the amino acid sequence SEQ ID NO:24.
在一些实施例中，所述抗C5a抗体包含VH，其包含如氨基酸序列SEQ ID NO:34所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:44所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3。In some embodiments, the anti-C5a antibody comprises a VH comprising HC-CDR1, HC-CDR2, and HC-CDR3 as shown in the amino acid sequence of SEQ ID NO:34; and a VL comprising VL as shown in the amino acid sequence SEQ ID NO: 44 includes LC-CDR1, LC-CDR2 and LC-CDR3.
在一些实施例中，所述抗C5a抗体包含：VH，所述VH包含氨基酸序列SEQ ID NO:34或其变体，所述变体与氨基酸序列SEQ ID NO:34具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及VL，所述VL包含氨基酸序列SEQ ID NO:44或其变体，所述变体与氨基酸序列SEQ ID NO:44具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述抗C5a抗体包含VH，所述VH包含氨基酸序列SEQ ID NO:34，以及VL，所述VL包含氨基酸序列SEQ ID NO:44。 In some embodiments, the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 34 or a variant thereof that is at least about 80% identical to the amino acid sequence SEQ ID NO: 34 (e.g., at least 80%, 85%, 90%, 95%, 96% , 97%, 98%, or 99%) sequence identity; and VL comprising the amino acid sequence SEQ ID NO: 44 or a variant thereof A variant having at least about 80% (eg, at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to the amino acid sequence SEQ ID NO: 44. In some embodiments, the anti-C5a antibody comprises a VH comprising the amino acid sequence SEQ ID NO:34, and a VL comprising the amino acid sequence SEQ ID NO:44.
在一些实施例中，上述氨基酸取代限于本文表4中所示的“示例性取代”。在一些实施例中，氨基酸取代限于本文表4中所示的“优选取代”。In some embodiments, the above-described amino acid substitutions are limited to the "exemplary substitutions" set forth in Table 4 herein. In some embodiments, amino acid substitutions are limited to the "preferred substitutions" set forth in Table 4 herein.
在一些实施例中，功能性表位可通过组合丙氨酸扫描法来解析。在此过程中，组合丙氨酸扫描技术可用于鉴定C5a蛋白中与抗C5a抗体相互作用所必需的氨基酸。在一些实施例中，该表位是构象的，同时可以采用与C5a蛋白结合的抗C5a抗体的晶体结构来鉴定表位。In some embodiments, functional epitopes can be resolved by combined alanine scanning methods. In this process, combinatorial alanine scanning techniques can be used to identify amino acids in the C5a protein that are necessary for interaction with anti-C5a antibodies. In some embodiments, the epitope is conformational and the crystal structure of an anti-C5a antibody bound to the C5a protein can be used to identify the epitope.
在一些实施例中，本申请提供与本文所述的任一种抗C5a抗体竞争性结合C5a的抗体。在一些实施例中，提供能够与本文所述的任一种抗C5a抗体竞争性地结合C5a上的表位的抗体。在一些实施例中，提供抗C5a抗体，其与包含VH和VL的抗C5a抗体分子结合相同的表位，其中所述VH包含SEQ ID NOs:25-34中任一所示的氨基酸序列，以及所述VL包含SEQ ID NOs:35-44中任一所示的氨基酸序列。在一些实施例中，提供抗C5a抗体，其与包含VH和VL的抗C5a抗体竞争性地结合C5a，其中所述VH包含SEQ ID NOs:25-34中任一所示的氨基酸序列，以及所述VL包含SEQ ID NOs:35-44中任一所示的氨基酸序列。In some embodiments, the application provides antibodies that compete for binding to C5a with any of the anti-C5a antibodies described herein. In some embodiments, antibodies are provided that are capable of binding to an epitope on C5a competitively with any of the anti-C5a antibodies described herein. In some embodiments, an anti-C5a antibody is provided that binds to the same epitope as an anti-C5a antibody molecule comprising VH and VL , wherein the VH comprises the amino acid set forth in any of SEQ ID NOs: 25-34 sequence, and the V L comprises the amino acid sequence shown in any one of SEQ ID NOs: 35-44. In some embodiments, an anti-C5a antibody is provided that competes for binding to C5a with an anti-C5a antibody comprising VH and VL , wherein the VH comprises the amino acid sequence set forth in any one of SEQ ID NOs: 25-34 , and the V L includes the amino acid sequence shown in any one of SEQ ID NOs: 35-44.
在一些实施例中，可以利用竞争实验来鉴定与本文所述的抗C5a抗体竞争性结合C5a的单克隆抗体。竞争实验可以通过识别相同的或空间上重叠的表位或者通过一个抗体竞争性抑制另一抗体与抗原结合来确定两个抗体是否结合相同的表位。在某些实施例中，这种竞争性抗体与本文所述的抗体结合相同的表位。一些示例性的竞争实验包括，但不限于如Harlow and Lane(1988)Antibodies:A Laboratory Manual ch.14(Cold Spring Harbor Laboratory，Cold Spring Harbor,N.Y.)中所提到的常规实验。用于解析抗体结合的表位的详细示例性方法如Morris(1996)"Epitope Mapping Protocols,"in Methods in Molecular Biology vol.66(Humana Press,Totowa,N.J.)中所述。在一些实施例中，如果每种抗体阻断另一种抗体结合的50％或更多，则称其结合相同的表位。在一些实施例中，与本文所述的抗C5a抗体竞争的抗体是嵌合抗体、人源化抗体或全人抗体。In some embodiments, competition experiments can be used to identify monoclonal antibodies that compete with the anti-C5a antibodies described herein for binding to C5a. Competition experiments can determine whether two antibodies bind the same epitope by identifying the same or spatially overlapping epitope or by one antibody competitively inhibiting the binding of another antibody to the antigen. In certain embodiments, such competing antibodies bind the same epitope as the antibodies described herein. Some exemplary competition experiments include, but are not limited to, conventional experiments as mentioned in Harlow and Lane (1988) Antibodies: A Laboratory Manual ch.14 (Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.). Detailed exemplary methods for resolving epitopes bound by antibodies are described in Morris (1996) "Epitope Mapping Protocols," in Methods in Molecular Biology vol. 66 (Humana Press, Totowa, N.J.). In some embodiments, each antibody is said to bind the same epitope if it blocks 50% or more of the binding of the other antibody. In some embodiments, the antibody that competes with an anti-C5a antibody described herein is a chimeric antibody, a humanized antibody, or a fully human antibody.
示例性抗C5a抗体序列如表2、表3所示，其中根据Kabat定义方式进行CDR编号。本领域技术人员将认识到有多种已知算法来预测CDR的位置以及界定抗体轻、重链可变区。包含如本文所述抗体的CDRs、VH和/或VL序列，但基于预测算法而非下表中所示例的抗体也在本申请的范围内。Exemplary anti-C5a antibody sequences are shown in Table 2 and Table 3, where CDR numbering is performed according to Kabat definition. Those skilled in the art will recognize that there are a variety of known algorithms for predicting the location of CDRs and defining antibody light and heavy chain variable regions. Antibodies containing the CDRs, VH and/or VL sequences of antibodies as described herein, but based on prediction algorithms other than those exemplified in the table below are also within the scope of the present application.
表2示例性抗C5a抗体CDR序列



Table 2 Exemplary anti-C5a antibody CDR sequences



表3示例性序列




Table 3 Exemplary sequences




C5aC5a
补体系统由30多种蛋白质组成，其响应组织损伤、病原体或其他异物入侵而激活。补体成分5a(C5a)最早被描述为具有趋化和过敏毒素性质的补体因子5(C5)的裂解产物(Shin等，1968)。C5a的前体C5蛋白含有1676个氨基酸，分子量为188kDa，其基因位于9q33–9q34(Wetsel等，1988)。人的C5a是由C5转化酶切割C5的α链所产生的一个约11kDa，含有74个氨基酸的糖蛋白，但N-连接的糖基化对其功能不是必需的。C5a的性质表明其是先天免疫反应的重要组成部分，但现有证据表明，C5a也可能在适应性免疫中起作用(Kohl，2006)。尽管C5a不一定是炎症性疾病的引发因素，但在许多炎症性疾病中会产生过量或不可控的C5a，这表明C5a可促进并维持炎症反应(Guo和Ward，2005年)。C5a具有四个采用肽环连接的反平行α螺旋，并通过三个关键的二硫键使其更稳定(Monk等，2007)。诱变和抗体研究已经确定了几个提供与受体相互作用的基本残基(Monk等，2007年综述)。The complement system is composed of more than 30 proteins that are activated in response to tissue damage, invasion by pathogens or other foreign bodies. Complement component 5a (C5a) was first described as a cleavage product of complement factor 5 (C5) with chemotactic and anaphylatoxin properties (Shin et al., 1968). The precursor of C5a, C5 protein, contains 1676 amino acids, has a molecular weight of 188 kDa, and its gene is located at 9q33–9q34 (Wetsel et al., 1988). Human C5a is an approximately 11 kDa glycoprotein containing 74 amino acids produced by C5 convertase cleavage of the α chain of C5, but N-linked glycosylation is not essential for its function. The properties of C5a suggest that it is an important component of the innate immune response, but existing evidence suggests that C5a may also play a role in adaptive immunity (Kohl, 2006). Although C5a is not necessarily a trigger of inflammatory diseases, excessive or uncontrolled C5a is produced in many inflammatory diseases, suggesting that C5a can promote and maintain inflammatory responses (Guo and Ward, 2005). C5a has four antiparallel α-helices connected by peptide loops and is made more stable by three key disulfide bonds (Monk et al., 2007). Mutagenesis and antibody studies have identified several essential residues that provide interaction with the receptor (reviewed by Monk et al., 2007).
补体级联可通过四种途径激活：经典途径，旁路途径，甘露聚糖结合凝集素途径(MBL)或外源蛋白酶途径(Ricklin和Lambris，2007)。经典途径和凝集素途径分别通过识别病原体表面上形成的抗体复合物以及细菌表面上的甘露糖而被激活。两种途径均导致丝氨酸蛋白酶切割C4，形成C4a和C4b。C4b结合C2后，在蛋白酶的作用下导致C2a的生成，C4b和C2a形成经典途径中的C3转化酶(C4b2a)。旁路途径可通过异物表面或“慢速转运”来激活，造成C3的自发水解，随后与因子B结合并形成了旁路途径中的C3转化酶(C3bBb)，旁路途径持续维持着低水平的补体级联激活，从而确保能对入侵的病原体作出快速响应(Ricklin和Lambris，2007)。以上三个途径都可形成C3转化酶，而C3转化酶进一步切割C3蛋白，形成C3a和C3b。C3b可促进细胞对病原体表面的识别以及病原体的清除，也可与C3转化酶(C4b2a或C3bBb)形成C5转化酶(C4b2aC3b或C3bBbC3b)，随后C5转化酶进一步切割C5产生C5a和C5b。C5b继续启动攻膜复合物(MAC；C5b-9)的组装。补体级联受到一系列可溶性的膜结合调节蛋白的紧密调节，这些蛋白阻止补体活化产物靶向宿主组织(Ricklin和Lambris，2007)。但是可以通过一些外源途径绕开这种控制，如凝血酶可直接切割C3和C5，造成补体系统的激活(Amara等，2008)。此外，活化的嗜中性粒细胞和肺泡巨噬细胞可通过分泌的丝氨酸蛋白酶，切割C5产生C5a(Amara等，2008)。血浆和细胞表面的羧肽酶能去除蛋白C端的精氨酸，因此经C5裂解产生的C5a可被其迅速代谢，形成C5adesArg(Bokisch和Muller-Eberhard，1970)。与C5a相比，C5adesArg的效力降低，致使其与经典C5a受体-CD88的结合亲和力降低(Higginbottom等，2005)。C5a和C5adesArg在体内能被迅速清除，其中约有50％的C5a和C5adesArg在2-3分钟内被从循环中清除，此外部分C5a与白细胞和其他细胞上的CD88结合后被介导清除(Oppermann和Gotze，1994)。但是，第二种受体，C5a样受体2(C5L2)可以更有效地去除补体片段，通过快速内化C5a/C5adesArg，特别是C5adesArg，使其保留其中并在某些细胞类型中降解(Scola等(2009)。相反，表达CD88的细胞将C5a内在化后，将C5a释放的比例更高，且是未降解的，可能仍处于活性状态的形式。血浆C5a也可以通过肝脏清除(Chenoweth和Goodman，1983)。The complement cascade can be activated through four pathways: the classical pathway, the alternative pathway, the mannan-binding lectin pathway (MBL) or the exogenous protease pathway (Ricklin and Lambris, 2007). The classical pathway and the lectin pathway are activated by recognition of antibody complexes formed on the surface of pathogens and mannose on bacterial surfaces, respectively. Both pathways result in cleavage of C4 by serine proteases, forming C4a and C4b. After C4b binds to C2, it leads to the generation of C2a under the action of protease. C4b and C2a form the C3 convertase (C4b2a) in the classical pathway. The alternative pathway can be activated by foreign body surfaces or "slow transport", resulting in spontaneous hydrolysis of C3, which subsequently binds to factor B and forms C3 convertase (C3bBb) in the alternative pathway, which continues to maintain low levels Activation of the complement cascade, thereby ensuring a rapid response to invading pathogens (Ricklin and Lambris, 2007). The above three pathways can form C3 convertase, and C3 convertase further cleaves the C3 protein to form C3a and C3b. C3b can promote cell recognition of pathogen surfaces and clearance of pathogens, and can also form C5 convertase (C4b2aC3b or C3bBbC3b) with C3 convertase (C4b2a or C3bBb), and then C5 convertase further cleaves C5 to produce C5a and C5b. C5b proceeds to initiate the assembly of the membrane attack complex (MAC; C5b-9). The complement cascade is tightly regulated by a series of soluble membrane-bound regulatory proteins that prevent complement activation products from targeting host tissues (Ricklin and Lambris, 2007). However, this control can be circumvented through some exogenous pathways. For example, thrombin can directly cleave C3 and C5, causing activation of the complement system (Amara et al., 2008). In addition, activated neutrophils and alveolar macrophages can cleave C5 to produce C5a through secreted serine proteases (Amara et al., 2008). Carboxypeptidase on the plasma and cell surface can remove the arginine at the C-terminus of the protein, so the C5a produced by C5 cleavage can be rapidly metabolized by it to form C5adesArg (Bokisch and Muller-Eberhard, 1970). Compared with C5a, C5adesArg has reduced potency, resulting in reduced binding affinity to the classical C5a receptor CD88 (Higginbottom et al., 2005). C5a and C5adesArg can be cleared rapidly in the body. About 50% of C5a and C5adesArg are cleared from the circulation within 2-3 minutes. In addition, some C5a is mediated after binding to CD88 on leukocytes and other cells (Oppermann and Gotze, 1994). However, a second receptor, C5a-like receptor 2 (C5L2), can more efficiently remove complement fragments by rapidly internalizing C5a/C5adesArg and, in particular, C5adesArg, allowing it to be retained and degraded in certain cell types (Scola (2009). In contrast, CD88-expressing cells internalize C5a and release a higher proportion of C5a in an undegraded, presumably active form. Plasma C5a can also be cleared by the liver (Chenoweth and Goodman , 1983).
CD88CD88
C5a以相似的高亲和力结合CD88和C5L2。相反的，C5adesArg对C5L2的亲和力与C5a相似(12nM)，但与CD88的亲和力却低得多(660nM)(Monk等，2007)。CD88和C5L2具有35％的序列同源性，并且位于19号染色体的同一区域(19q13.3–19q13.4)。它们与其他趋化因子受体(例如甲酰基肽受体家族和缓激肽受体)基因聚集成簇。两者都是糖基化的七次跨膜蛋白，分子量约为45kDa。CD88是一个G蛋白偶联受体，同时也是视紫红质基因家族的成员之一(Monk等，2007)。人们认为C5a与CD88的结合发生在两个不同且物理上分离的位点。第一个“识别”位点位于受体的细胞外氨基末端(N末端)，其与C5aN端和二硫键连接的核心结合。第二个“激活”位点由受体的跨膜结构域形成，其与C5a的C末端结合，并导致由受体偶联的G蛋白介导的特定信号转导途径产生(Monk等，2007)。C5a binds CD88 and C5L2 with similarly high affinity. In contrast, the affinity of C5adesArg for C5L2 is similar to that of C5a ( 12nM), but has a much lower affinity for CD88 ( 660 nM) (Monk et al., 2007). CD88 and C5L2 share 35% sequence homology and are located in the same region of chromosome 19 (19q13.3–19q13.4). They cluster with other chemokine receptor genes such as the formyl peptide receptor family and bradykinin receptors. Both are glycosylated seven-transmembrane proteins with a molecular weight of approximately 45 kDa. CD88 is a G protein-coupled receptor and a member of the rhodopsin gene family (Monk et al., 2007). Binding of C5a to CD88 is thought to occur at two distinct and physically separate sites. The first "recognition" site is located at the extracellular amino terminus (N-terminus) of the receptor, which binds to the C5a N-terminus and disulfide-linked core. The second “activation” site is formed by the transmembrane domain of the receptor, which binds to the C-terminus of C5a and leads to the generation of a specific signaling pathway mediated by the receptor-coupled G protein (Monk et al., 2007 ).
C5L2C5L2
表达C5L2的细胞类型与表达CD88的细胞类型大致相同，例如嗜中性粒细胞，单核细胞，淋巴细胞，巨噬细胞，以及非骨髓细胞(例如血管平滑肌细胞)和组织来源的细胞(例如肾上腺、心脏、肝、肺脾和大脑)，但在非炎性条件下，C5L2的表达含量却显著低于CD88的表达含量(Gao等，2005)。C5L2的功能尚不清楚。一些实验数据表明，C5L2可作为无信号转导功能的诱饵受体。研究发现敲除或阻断C5L2会加剧小鼠的炎症反应(Gao等，2005；Gerard等，2005)。这表明C5L2具有抗炎功能，其可能通过减少能与CD88结合的C5a的量起作用。此外，C5L2可充当正调节剂，至少在小鼠中，其对C5a和C3a信号传导至关重要(Chen等人，2007)。在体外，研究发现C5L2对促进嗜中性粒细胞，巨噬细胞和成纤维细胞中的C5a信号传导至关重要，而体内缺乏C5L2可导致卵白蛋白诱导的气道高反应性和炎症(Chen等，2007)。此外，在小鼠“晚期”败血症(100％致死率)模型中，只有同时阻滞C5L2和CD88才可起到保护作用(Rittirsch等，2008)。Cell types that express C5L2 are broadly the same as those that express CD88, such as neutrophils, monocytes, lymphocytes, macrophages, as well as non-myeloid cells such as vascular smooth muscle cells and cells of tissue origin such as the adrenal gland. , heart, liver, lung, spleen and brain), but under non-inflammatory conditions, the expression level of C5L2 is significantly lower than that of CD88 (Gao et al., 2005). The function of C5L2 is unknown. Some experimental data suggest that C5L2 may serve as a decoy receptor without signal transduction function. Studies have found that knocking out or blocking C5L2 can exacerbate the inflammatory response in mice (Gao et al., 2005; Gerard et al., 2005). This suggests that C5L2 has anti-inflammatory functions, possibly by reducing the amount of C5a that can bind to CD88. Furthermore, C5L2 can act as a positive regulator that is critical for C5a and C3a signaling, at least in mice (Chen et al., 2007). In vitro, C5L2 was found to be critical for promoting C5a signaling in neutrophils, macrophages, and fibroblasts, whereas lack of C5L2 in vivo resulted in ovalbumin-induced airway hyperresponsiveness and inflammation (Chen et al. , 2007). Furthermore, in a mouse model of “late” sepsis (100% lethality), only simultaneous blockade of C5L2 and CD88 was protective (Rittirsch et al., 2008).
全长抗C5a抗体Full-length anti-C5a antibody
在一些实施例中，所述抗C5a抗体是全长抗C5a抗体。在一些实施例中，所述全长抗C5a抗体是IgA、IgD、IgE、IgG或IgM。在一些实施例中，所述全长抗C5a抗体包含IgG恒定区域，例如IgG1、IgG2、IgG3、IgG4或其变体的恒定区域。在一些实施例中，所述全长抗C5a抗体包含λ轻链恒定区。在一些实施例中，所述全长抗C5a抗体包含κ轻链恒定区。在一些实施例中，所述全长抗C5a抗体是全长的人抗C5a抗体。在一些实施例中，所述全长抗C5a抗体包含小鼠免疫球蛋白Fc序列。在一些实施例中，所述全长抗C5a抗体包含已经改变的或以其他方式改变的Fc序列，使得其具有增强的抗体依赖的细胞介导的细胞毒作用(ADCC)和补体依赖的细胞毒作用(CDC)的效应功能。In some embodiments, the anti-C5a antibody is a full-length anti-C5a antibody. In some embodiments, the full-length anti-C5a antibody is IgA, IgD, IgE, IgG, or IgM. In some embodiments, the full-length anti-C5a antibody comprises an IgG constant region, such as that of IgG1, IgG2, IgG3, IgG4, or a variant thereof. In some embodiments, the full-length anti-C5a antibody comprises a lambda light chain constant region. In some embodiments, the full-length anti-C5a antibody comprises a kappa light chain constant region. In some embodiments, the full-length anti-C5a antibody is a full-length human anti-C5a antibody. In some embodiments, the full-length anti-C5a antibody comprises mouse immunoglobulin Fc sequence. In some embodiments, the full-length anti-C5a antibody comprises an Fc sequence that has been altered or otherwise altered such that it has enhanced antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity. Effector functions of (CDC).
因此，例如，在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，所述抗C5a抗体与C5a特异性结合。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。Thus, for example, in some embodiments, a full-length anti-C5a antibody is provided comprising an IgG1 constant region that specifically binds to C5a. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG2恒定区的全长抗C5a抗体，所述抗C5a抗体与C5a特异性结合。在一些实施例中，所述IgG2是人IgG2。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG2 constant region is provided, the anti-C5a antibody specifically binding to C5a. In some embodiments, the IgG2 is human IgG2. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
在一些实施例中，提供一种包含IgG3恒定区的全长抗C5a抗体，所述抗C5a抗体与C5a特异性结合。在一些实施例中，所述IgG3是人IgG3。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG3 constant region is provided, the anti-C5a antibody specifically binding to C5a. In some embodiments, the IgG3 is human IgG3. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，所述抗C5a抗体与C5a特异性结合。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, the anti-C5a antibody specifically binding to C5a. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含SEQ ID NO:1所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；HC-CDR2，其包含SEQ ID NOs:2-5中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；和HC-CDR3，其包含SEQ ID NOs:6-11中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含SEQ ID NOs:12-16中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代，LC-CDR2，其包含SEQ ID NOs:17-19中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；和LC-CDR3，其包含SEQ ID NOs:20-24中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO: 1 or a variant thereof, which variant includes substitutions of up to about 3 (e.g., 1, 2 or 3) amino acids; HC-CDR2, which includes SEQ ID NOs: The amino acid sequence shown in any one of 2-5, or a variant thereof, the variant comprising a substitution of up to about 3 (e.g., 1, 2 or 3) amino acids; and HC-CDR3, which comprises SEQ ID NOs: 6 - the amino acid sequence shown in any one of 11 or a variant thereof, the variant comprising a substitution of up to about 3 (eg 1, 2 or 3) amino acids; and b) a light chain variable domain, the light chain variable domain The chain variable domain includes: LC-CDR1, which includes the amino acid sequence shown in any one of SEQ ID NOs: 12-16 or a variant thereof, which variant includes up to about 3 (e.g., 1, 2 or 3 ) amino acid substitutions, LC-CDR2, which comprise the amino acid sequence shown in any one of SEQ ID NOs: 17-19 or a variant thereof, the variant comprising up to about 3 (such as 1, 2 or 3) amino acids Substitutions; and LC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs: 20-24 or a variant thereof, said variant comprising up to about 3 (e.g., 1, 2 or 3) amino acids replace. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG2恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含SEQ ID NO:1所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；HC-CDR2，其包含SEQ ID NOs:2-5中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；和HC-CDR3，其包含SEQ ID NOs:6-11中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含SEQ ID NOs:12-16中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；LC-CDR2，其包含SEQ ID NOs:17-19中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；和LC-CDR3，其包含SEQ ID NOs:20-24中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代。在一些实施例中，所述IgG2是人IgG2。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG2 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO: 1 or a variant thereof, which variant includes substitutions of up to about 3 (e.g., 1, 2 or 3) amino acids; HC-CDR2, which includes SEQ ID NOs: The amino acid sequence shown in any one of 2-5, or a variant thereof, the variant comprising a substitution of up to about 3 (e.g., 1, 2 or 3) amino acids; and HC-CDR3, which comprises SEQ ID NOs: 6 - the amino acid sequence shown in any one of 11 or a variant thereof, the variant comprising a substitution of up to about 3 (eg 1, 2 or 3) amino acids; and b) a light chain variable domain, the light chain variable domain The chain variable domain includes: LC-CDR1, which includes the amino acid sequence shown in any one of SEQ ID NOs: 12-16 or a variant thereof, which variant includes up to about 3 (e.g., 1, 2 or 3 ) Substitution of amino acids; LC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs: 17-19 or a variant thereof, the variant comprising up to about 3 (e.g. 1, 2 or 3) amino acids Substitutions; and LC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs: 20-24 or a variant thereof, said variant comprising up to about 3 (e.g., 1, 2 or 3) amino acids replace. In some embodiments, the IgG2 is human IgG2. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
在一些实施例中，提供一种包含IgG3恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含SEQ ID NO:1所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代，HC-CDR2，其包含SEQ ID NOs:2-5中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；和HC-CDR3，其包含SEQ ID NOs:6-11中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含SEQ ID NOs:12-16中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；LC-CDR2，其包含SEQ ID NOs:17-19中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；和LC-CDR3，其包含SEQ ID NOs:20-24中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代。在一些实施例中，所述IgG3是人IgG3。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG3 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO: 1 or a variant thereof, which variant includes a substitution of up to about 3 (e.g., 1, 2 or 3) amino acids, HC-CDR2, which includes SEQ ID NOs: The amino acid sequence shown in any one of 2-5, or a variant thereof, the variant comprising a substitution of up to about 3 (e.g., 1, 2 or 3) amino acids; and HC-CDR3, which comprises SEQ ID NOs: 6 - the amino acid sequence shown in any one of 11 or a variant thereof, the variant comprising a substitution of up to about 3 (eg 1, 2 or 3) amino acids; and b) a light chain variable domain, the light chain variable domain The chain variable domain includes: LC-CDR1, which includes the amino acid sequence shown in any one of SEQ ID NOs: 12-16 or a variant thereof, which variant includes up to about 3 (e.g., 1, 2 or 3 ) Substitution of amino acids; LC-CDR2 comprising the amino acid sequence shown in any one of SEQ ID NOs: 17-19 or a variant thereof, the variant comprising up to about 3 (e.g. 1, 2 or 3) amino acids Substitutions; and LC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs: 20-24 or a variant thereof, said variant comprising up to about 3 (e.g., 1, 2 or 3) amino acids replace. In some embodiments, the IgG3 is human IgG3. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含SEQ ID NO:1所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；HC-CDR2，其包含SEQ ID NOs:2-5中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；和HC-CDR3，其包含SEQ ID NOs:6-11中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含SEQ ID NOs:12-16中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代，LC-CDR2，其包含SEQ ID NOs:17-19中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代，和LC-CDR3，其包含SEQ ID NOs:20-24中任一所示的氨基酸序列或其变体，所述变体包含至多约3个(例如1、2或3个)氨基酸的取代。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO: 1 or a variant thereof, which variant includes substitutions of up to about 3 (e.g., 1, 2 or 3) amino acids; HC-CDR2, which includes SEQ ID NOs: The amino acid sequence shown in any one of 2-5, or a variant thereof, the variant comprising a substitution of up to about 3 (e.g., 1, 2 or 3) amino acids; and HC-CDR3, which comprises SEQ ID NOs: 6 - the amino acid sequence shown in any one of 11 or a variant thereof, the variant comprising a substitution of up to about 3 (eg 1, 2 or 3) amino acids; and b) a light chain variable domain, the light chain variable domain The chain variable domain includes: LC-CDR1, which includes the amino acid sequence shown in any one of SEQ ID NOs: 12-16 or a variant thereof, which variant includes up to about 3 (e.g., 1, 2 or 3 ) amino acid substitutions, LC-CDR2, which comprise the amino acid sequence shown in any one of SEQ ID NOs: 17-19 or a variant thereof, the variant comprising up to about 3 (such as 1, 2 or 3) amino acids substitutions, and LC-CDR3 comprising the amino acid sequence shown in any one of SEQ ID NOs: 20-24 or a variant thereof, the variant comprising up to about 3 (e.g., 1, 2 or 3) amino acids replace. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含SEQ ID NO:1所示的氨基酸序列，HC-CDR2，其包含SEQ ID NOs:2-5中任一所示的氨基酸序列，和HC-CDR3，其包含SEQ ID NOs:6-11中任一所示的氨基酸序列，或者所述重链可变结构域的变体，其HC-CDR序列中包含至多约5个(例如1、2、3、4或5个)氨基酸的取代；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含SEQ ID NOs:12-16中任一所示的氨基酸序列，LC-CDR2，其包含SEQ ID NOs:17-19中任一所示的氨基酸序列，和LC-CDR3，其包含SEQ ID NOs:20-24中任一所示的氨基酸序列，或者所述轻链可变结构域的变体，其LC-CDR序列中包含至多约5个(例如1、2、3、4或5个)氨基酸的取代。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO:1, HC-CDR2, which includes the amino acid sequence shown in any one of SEQ ID NOs:2-5, and HC-CDR3, which includes SEQ ID NOs:6-11 The amino acid sequence shown in any one of them, or a variant of the heavy chain variable domain, the HC-CDR sequence of which contains at most about 5 (such as 1, 2, 3, 4 or 5) amino acid substitutions; and b) a light chain variable domain comprising: LC-CDR1 comprising the amino acid sequence shown in any one of SEQ ID NOs: 12-16, LC-CDR2 comprising SEQ ID The amino acid sequence shown in any one of NOs: 17-19, and LC-CDR3, which includes the amino acid sequence shown in any one of SEQ ID NOs: 20-24, or a variant of the light chain variable domain, The LC-CDR sequence contains up to about 5 (eg, 1, 2, 3, 4 or 5) amino acid substitutions. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含SEQ ID NO:1所示的氨基酸序列，HC-CDR2，其包含SEQ ID NOs:2-5中任一所示的氨基酸序列，和HC-CDR3，其包含SEQ ID NOs:6-11中任一所示的氨基酸序列，或者所述重链可变结构域的变体，其HC-CDR序列中包含至多约5个(例如1、2、3、4或5个)氨基酸的取代；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含SEQ ID NOs:12-16中任一所示的氨基酸序列，LC-CDR2，包含SEQ ID NOs:17-19中任一所示的氨基酸序列，和LC-CDR3，其包含SEQ ID NOs:20-24中任一所示的氨基酸序列，或者所述轻链可变结构域的变体，其LC-CDR序列中包含至多约5个(例如1、2、3、4或5个)氨基酸的取代。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises a) a heavy chain variable domain comprising: HC-CDR1, It includes the amino acid sequence shown in SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence shown in any of SEQ ID NOs: 2-5, and HC-CDR3, which includes the amino acid sequence shown in any of SEQ ID NOs: 6-11 Any of the amino acid sequences shown, or a variant of the heavy chain variable domain, comprising up to about 5 (e.g., 1, 2, 3, 4 or 5) amino acid substitutions in the HC-CDR sequence; and b) Light chain variable domain, the light chain variable domain comprising: LC-CDR1, comprising the amino acid sequence shown in any one of SEQ ID NOs: 12-16, LC-CDR2, comprising SEQ ID NOs: The amino acid sequence shown in any one of 17-19, and LC-CDR3, which includes the amino acid sequence shown in any one of SEQ ID NOs: 20-24, or a variant of the light chain variable domain whose LC - Substitutions comprising up to about 5 (eg 1, 2, 3, 4 or 5) amino acids in the CDR sequence. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含SEQ ID NO:1所示的氨基酸序列，HC-CDR2，其包含SEQ ID NOs:2-5中任一所示的氨基酸序列，和HC-CDR3，其包含SEQ ID NOs:6-11中任一所示的氨基酸序列；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含SEQ ID NOs:12-16中任一所示的氨基酸序列，LC-CDR2，其包含SEQ ID NOs:17-19中任一所示的氨基酸序列，和LC-CDR3，其包含SEQ ID NOs:20-24中任一所示的氨基酸序列。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO:1, HC-CDR2, which includes the amino acid sequence shown in any one of SEQ ID NOs:2-5, and HC-CDR3, which includes SEQ ID NOs:6-11 The amino acid sequence shown in any one of; and b) a light chain variable domain, the light chain variable domain comprising: LC-CDR1, which includes the amino acid sequence shown in any one of SEQ ID NOs: 12-16 , LC-CDR2, which includes the amino acid sequence shown in any one of SEQ ID NOs: 17-19, and LC-CDR3, which includes the amino acid sequence shown in any one of SEQ ID NOs: 20-24. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含SEQ ID NO:1所示的氨基酸序列，HC-CDR2，其包含SEQ ID NOs:2-5中任一所示的氨基酸序列，和HC-CDR3，其包含SEQ ID NOs:6-11中任一所示的氨基酸序列；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含SEQ ID NOs:12-16中任一所示的氨基酸序列，LC-CDR2，其包含SEQ ID NOs:17-19中任一所示的氨基酸序列，和LC-CDR3，其包含SEQ ID NOs:20-24中任一所示的氨基酸序列。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence shown in SEQ ID NO:1, HC-CDR2, which includes the amino acid sequence shown in any one of SEQ ID NOs:2-5, and HC-CDR3, which includes SEQ ID NOs:6-11 The amino acid sequence shown in any one of; and b) a light chain variable domain, the light chain variable domain comprising: LC-CDR1, which includes the amino acid sequence shown in any one of SEQ ID NOs: 12-16 , LC-CDR2, which includes the amino acid sequence shown in any one of SEQ ID NOs: 17-19, and LC-CDR3, which includes the amino acid sequence shown in any one of SEQ ID NOs: 20-24. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:2，和HC-CDR3，其包含氨基酸序列SEQ ID NO:6；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 2, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 6; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :20. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :20. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:8；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:13，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:21。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 8; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 13, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty one. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:5，和HC-CDR3，其包含氨基酸序列SEQ ID NO:10；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 5, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 10; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:23。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty three. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:15，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 15, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:11；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:16，LC-CDR2，其包含氨基酸序列SEQ ID NO:19，和LC-CDR3，其包含氨基酸序列SEQ ID NO:24。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 11; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 16, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 19, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty four. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:2，和HC-CDR3，其包含氨基酸序列SEQ ID NO:6；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 2, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 6; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :20. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :20. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:8；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:13，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:21。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 8; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 13, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty one. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:5，和HC-CDR3，其包含氨基酸序列SEQ ID NO:10；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 5, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 10; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:23。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 12, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty three. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:15，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 15, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 14, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty two. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中所述抗C5a抗体包含：a)重链可变结构域，所述重链可变结构域包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:11；以及b)轻链可变结构域，所述轻链可变结构域包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:16，LC-CDR2，其包含氨基酸序列SEQ ID NO:19，和LC-CDR3，其包含氨基酸序列SEQ ID NO:24。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a) a heavy chain variable domain, the heavy chain variable domain comprising: HC-CDR1 , which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 11; and b) a light chain variable domain, The light chain variable domain includes: LC-CDR1, which includes the amino acid sequence SEQ ID NO: 16, LC-CDR2, which includes the amino acid sequence SEQ ID NO: 19, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: :twenty four. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中抗C5a抗体包含：重链可变结构域(VH)，所述VH包含SEQ ID NOs:25-34中任一所示的氨基酸序列或其变体，所述变体与SEQ ID NOs:25-34中任一所示的氨基酸序列具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及轻链可变结构域(VL)，所述VL包含SEQ ID NOs:35-44中任一所示的氨基酸序列或其变体，所述变体与SEQ ID NOs:35-44中任一所示的氨基酸序列具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: a heavy chain variable domain ( VH ), the VH comprising SEQ ID NOs: 25-34 Any amino acid sequence shown or a variant thereof, which variant has at least about 80% (such as at least 80%, 85%, 90%, 95%) with the amino acid sequence shown in any one of SEQ ID NOs: 25-34 %, 96%, 97%, 98% or 99% ) sequence identity; and a light chain variable domain ( VL ) comprising the amino acid sequence shown in any one of SEQ ID NOs: 35-44 Or a variant thereof that shares at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%) with the amino acid sequence shown in any one of SEQ ID NOs: 35-44 , 98% or 99%) sequence identity. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG2恒定区的全长抗C5a抗体，其中抗C5a抗体包含：重链可变结构域(VH)，所述VH包含SEQ ID NOs:25-34中任一所示的氨基酸序列或其变体，所述变体与SEQ ID NOs:25-34中任一所示的氨基酸序列具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及轻链可变结构域(VL)，所述VL包含SEQ ID NOs:35-44中任一所示的氨基酸序列或其变体，所述变体与SEQ ID NOs:35-44中任一所示的氨基酸序列具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述IgG2是人IgG2。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG2 constant region is provided, wherein the anti-C5a antibody comprises: a heavy chain variable domain ( VH ), the VH comprising SEQ ID NOs: 25-34 Any amino acid sequence shown or a variant thereof, which variant has at least about 80% (such as at least 80%, 85%, 90%, 95%) with the amino acid sequence shown in any one of SEQ ID NOs: 25-34 %, 96%, 97%, 98% or 99%) sequence identity; and a light chain variable domain ( VL ) comprising the amino acid sequence shown in any one of SEQ ID NOs: 35-44 Or a variant thereof that shares at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%) with the amino acid sequence shown in any one of SEQ ID NOs: 35-44 , 98% or 99%) sequence identity. In some embodiments, the IgG2 is human IgG2. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG3恒定区的全长抗C5a抗体，其中抗C5a抗体包含：重链可变结构域(VH)，所述VH包含SEQ ID NOs:25-34中任一所示的氨基酸序列或其变体，所述变体与SEQ ID NOs:25-34中任一所示的氨基酸序列具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及轻链可变结构域(VL)，所述VL包含SEQ ID NOs:35-44中任一所示的氨基酸序列或其变体，所述变体与SEQ ID NOs:35-44中任一所示的氨基酸序列具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述IgG3是人IgG3。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG3 constant region is provided, wherein the anti-C5a antibody comprises: a heavy chain variable domain ( VH ), the VH comprising SEQ ID NOs: 25-34 Any amino acid sequence shown or a variant thereof, which variant has at least about 80% (such as at least 80%, 85%, 90%, 95%) with the amino acid sequence shown in any one of SEQ ID NOs: 25-34 %, 96%, 97%, 98% or 99% ) sequence identity; and a light chain variable domain ( VL ) comprising the amino acid sequence shown in any one of SEQ ID NOs: 35-44 Or a variant thereof that shares at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%) with the amino acid sequence shown in any one of SEQ ID NOs: 35-44 , 98% or 99%) sequence identity. In some embodiments, the IgG3 is human IgG3. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中抗C5a抗体包含：重链可变结构域(VH)，所述VH包含SEQ ID NOs:25-34中任一所示的氨基酸序列或其变体，所述变体与SEQ ID NOs:25-34中任一所示的氨基酸序列具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性；以及轻链可变结构域(VL)，所述VL包含SEQ ID NOs:35-44中任一所示的氨基酸序列或其变体，所述变体与SEQ ID NOs:35-44中任一所示的氨基酸序列具有至少约80％(例如至少80％、85％、90％、95％、96％、97％、98％或99％)序列同一性。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a heavy chain variable domain ( VH ), the VH comprising SEQ ID NOs: 25-34 Any amino acid sequence shown or a variant thereof, which variant has at least about 80% (such as at least 80%, 85%, 90%, 95%) with the amino acid sequence shown in any one of SEQ ID NOs: 25-34 %, 96%, 97%, 98% or 99% ) sequence identity; and a light chain variable domain ( VL ) comprising the amino acid sequence shown in any one of SEQ ID NOs: 35-44 Or a variant thereof that shares at least about 80% (e.g., at least 80%, 85%, 90%, 95%, 96%, 97%) with the amino acid sequence shown in any one of SEQ ID NOs: 35-44 , 98% or 99%) sequence identity. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中抗C5a抗体包含：重链可变结构域(VH)，所述(VH)包含SEQ ID NOs:25-34中任一所示的氨基酸序列，以及轻链可变结构域(VL)，所述(VL)包含SEQ ID NOs:35-44中任一所示的氨基酸序列。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgGl constant region is provided, wherein the anti-C5a antibody comprises: a heavy chain variable domain ( VH ) comprising SEQ ID NOs: 25- The amino acid sequence shown in any one of SEQ ID NOs: 35-44, and the light chain variable domain (V L ), said (V L ) comprising the amino acid sequence shown in any one of SEQ ID NOs: 35-44. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中抗C5a抗体包含：重链可变结构域(VH)，所述VH包含SEQ ID NOs:25-34中任一所示的氨基酸序列，以及轻链可变结构域(VL)，所述VL包含SEQ ID NOs:35-44中任一所示的氨基酸序列。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: a heavy chain variable domain ( VH ), the VH comprising SEQ ID NOs: 25-34 The amino acid sequence shown in any one, and the light chain variable domain ( VL ), the VL comprising the amino acid sequence shown in any one of SEQ ID NOs: 35-44. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:25或其变体，所述变体与氨基酸序列SEQ ID NO:25具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:35或其变体，所述变体与氨基酸序列SEQ ID NO:35具有至少约80％序列同一性。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 25 or a variant thereof, the variant having the same amino acid sequence SEQ ID NO:25 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:35 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:35 Identity. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:26或其变体，所述变体与氨基酸序列SEQ ID NO:26具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:36或其变体，所述变体与氨基酸序列SEQ ID NO:36具有至少约80％序列同一性。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 26 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:26 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:36 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:36 Identity. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:27或其变体，所述变体与氨基酸序列SEQ ID NO:27具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:37或其变体，所述变体与氨基酸序列SEQ ID NO:37具有至少约80％序列同一性。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 27 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:27 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:37 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:37 Identity. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:28或其变体，所述变体与氨基酸序列SEQ ID NO:28具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:38或其变体，所述变体与氨基酸序列SEQ ID NO:38具有至少约80％序列同一性。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 28 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:28 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:38 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:38 Identity. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:29或其变体，所述变体与氨基酸序列SEQ ID NO:29具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:39或其变体，所述变体与氨基酸序列SEQ ID NO:39具有至少约80％序列同一性。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 29 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:29 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:39 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:39 Identity. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:30或其变体，所述变体与氨基酸序列SEQ ID NO:30具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:40或其变体，所述变体与氨基酸序列SEQ ID NO:40具有至少约80％序列同一性。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 30 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:30 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:40 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:40 Identity. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:31或其变体，所述变体与氨基酸序列SEQ ID NO:31具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:41或其变体，所述变体与氨基酸序列SEQ ID NO:41具有至少约80％序列同一性。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 31 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:31 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:41 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:41 Identity. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:32或其变体，所述变体与氨基酸序列SEQ ID NO:32具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:42或其变体，所述变体与氨基酸序列SEQ ID NO:42具有至少约80％序列同一性。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 32 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:32 having at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:42 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:42 Identity. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:33或其变体，所述变体与氨基酸序列SEQ ID NO:33具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:43或其变体，所述变体与氨基酸序列SEQ ID NO:43具有至少约80％序列同一性。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 33 or a variant thereof, the variant is identical to the amino acid sequence SEQ ID NO:33 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:43 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:43 Identity. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG1恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:34或其变体，所述变体与氨基酸序列SEQ ID NO:34具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:44或其变体，所述变体与氨基酸序列SEQ ID NO:44具有至少约80％序列同一性。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG1 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 34 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:34 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:44 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:44 Identity. In some embodiments, the IgG1 is human IgG1. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:25或其变体，所述变体与氨基酸序列SEQ ID NO:25具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:35或其变体，所述变体与氨基酸序列SEQ ID NO:35具有至少约80％序列同一性。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 25 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:25 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:35 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:35 Identity. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:26或其变体，所述变体与氨基酸序列SEQ ID NO:26具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:36或其变体，所述变体与氨基酸序列SEQ ID NO:36具有至少约80％序列同一性。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 26 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:26 having at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:36 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:36 Identity. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:27或其变体，所述变体与氨基酸序列SEQ ID NO:27具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:37或其变体，所述变体与氨基酸序列SEQ ID NO:37具有至少约80％序列同一性。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 27 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:27 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:37 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:37 Identity. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:28或其变体，所述变体与氨基酸序列SEQ ID NO:28具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:38或其变体，所述变体与氨基酸序列SEQ ID NO:38具有至少约80％序列同一性。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 28 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:28 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:38 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:38 Identity. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:29或其变体，所述变体与氨基酸序列SEQ ID NO:29具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:39或其变体，所述变体与氨基酸序列SEQ ID NO:39具有至少约80％序列同一性。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 29 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:29 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:39 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:39 Identity. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:30或其变体，所述变体与氨基酸序列SEQ ID NO:30具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:40或其变体，所述变体与氨基酸序列SEQ ID NO:40具有至少约80％序列同一性。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 30 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:30 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:40 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:40 Identity. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:31或其变体，所述变体与氨基酸序列SEQ ID NO:31具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:41或其变体，所述变体与氨基酸序列SEQ ID NO:41具有至少约80％序列同一性。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 31 or a variant thereof, the variant having the same amino acid sequence SEQ ID NO:31 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:41 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:41 Identity. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:32或其变体，所述变体与氨基酸序列SEQ ID NO:32具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:42或其变体，所述变体与氨基酸序列SEQ ID NO:42具有至少约80％序列同一性。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 32 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:32 having at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:42 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:42 Identity. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:33或其变体，所述变体与氨基酸序列SEQ ID NO:33具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:43或其变体，所述变体与氨基酸序列SEQ ID NO:43具有至少约80％序列同一性。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 33 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:33 has at least about 80% sequence identity; and VL comprising the amino acid sequence SEQ ID NO:43 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:43 Identity. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
在一些实施例中，提供一种包含IgG4恒定区的全长抗C5a抗体，其中抗C5a抗体包含：VH，其包含氨基酸序列SEQ ID NO:34或其变体，所述变体与氨基酸序列SEQ ID NO:34具有至少约80％序列同一性；以及VL，其包含氨基酸序列SEQ ID NO:44或其变体，所述变体与氨基酸序列SEQ ID NO:44具有至少约80％序列同一性。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成以及轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, a full-length anti-C5a antibody comprising an IgG4 constant region is provided, wherein the anti-C5a antibody comprises: V H comprising the amino acid sequence SEQ ID NO: 34 or a variant thereof, the variant being identical to the amino acid sequence SEQ ID NO:34 has at least about 80% sequence identity; and V L comprising the amino acid sequence SEQ ID NO:44 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence SEQ ID NO:44 Identity. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:46 and the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:48.
结合亲和力binding affinity
结合亲和力采用Kd、Koff、Kon或Ka表示。如本文所用，术语Koff是指抗体从抗原/抗体复合物中解离的速率常数，通过动力学选择装置测定。术语Kon是指抗体与抗原结合形成抗原/抗体复合物的结合速率常数。本文所用的平衡解离常数Kd是指特定抗体抗原相互作用时的解离常数，是指在抗体分子溶液中，抗原占据所有抗体结合位点的一半并且达到平衡时所需的抗原浓度，等于Koff/Kon。Kd的测定假设所有的结合分子均在溶液中。抗体与细胞壁连接的情况，例如在酵母表达系统中，相应的平衡解离速率常数采用EC50来表示，其是Kd的一个良好的近似值。亲和结合常数Ka是解离常数Kd的倒数。Binding affinity is expressed as Kd, Koff, Kon or Ka. As used herein, the term Koff refers to the rate constant for the dissociation of an antibody from an antigen/antibody complex, as measured by a kinetic selection device. The term Kon refers to the binding rate constant at which an antibody binds to an antigen to form an antigen/antibody complex. The equilibrium dissociation constant Kd used in this article refers to the dissociation constant when a specific antibody-antigen interacts. It refers to the antigen concentration required when the antigen occupies half of all antibody binding sites in the antibody molecule solution and reaches equilibrium, which is equal to Koff /Kon. The determination of Kd assumes that all bound molecules are in solution. In the case of antibodies bound to the cell wall, such as in yeast expression systems, the corresponding equilibrium dissociation rate constant is expressed as EC50, which is a good approximation of Kd. The affinity binding constant Ka is the reciprocal of the dissociation constant Kd.
解离常数(Kd)可以作为反应抗体部分与抗原亲和力的指标。例如，可以通过Scatchard方法使用标记有各种标记物的抗体，和Biacore仪器(由Amersham Biosciences制造)进行简单分析，根据用户手册或附带试剂盒，通过表面等离子体共振来分析生物分子间的相互作用。使用这些方法得到的Kd值，用单位M来表示。与靶标特异性结合的抗体可能具有，例如≤10-7M、≤10-8M、≤10-9M、≤10-10M、≤10-11M、≤10-12M或≤10-13M的Kd值。The dissociation constant (Kd) can be used as an indicator of the affinity of the reactive antibody moiety to the antigen. For example, a simple analysis can be performed by the Scatchard method using antibodies labeled with various markers and a Biacore instrument (manufactured by Amersham Biosciences) to analyze interactions between biomolecules by surface plasmon resonance according to the user manual or the included kit. . The Kd value obtained using these methods is expressed in the unit M. Antibodies that specifically bind a target may have, for example, ≤10 -7 M, ≤10 -8 M, ≤10 -9 M, ≤10 -10 M, ≤10 -11 M , ≤10 -12 M, or ≤10 - Kd value of 13 M.
抗体的结合特异性可以通过本领域已知的方法进行实验测定。这些方法包括，但不限于Western blots、ELISA-、RIA-、ECL-、IRMA-、EIA-、BIAcore测试和肽扫描等。The binding specificity of an antibody can be determined experimentally by methods known in the art. These methods include, but are not limited to, Western blots, ELISA-, RIA-, ECL-, IRMA-, EIA-, BIAcore testing, and peptide scanning.
在一些实施例中，所述抗C5a抗体特异性结合C5a靶标，其Kd值为10-7M至10-13M(例如10-7M至10-13M、10-8M至10-13M、10-9M至10-13M或10-10M至10-12M)。因此，在一些实施例中，抗C5a抗体与C5a之间结合的Kd值为10-7M至10-13M、1×10-7M至5×10-13M、10-7M至10-12M、10-7M至10-11M、10-7M至10-10M、10-7M至10-9M、10-8M至10-13M、1×10-8M至5×10-13M、10-8M至10-12M、10-8M至10-11M、10-8M至10-10M、10-8M至10-9M、5×10-9M至1×10-13M、5×10-9M至1×10-12M、5×10-9M至1×10-11M、5×10-9M至1×10-10M、10-9M至10-13M、10-9M至10-12M、10-9M至10-11M、10-9M至10-10M、5×10-10M至1×10-13M、5×10-10M至1×10- 12M、5×10-10M至1×10-11M、10-10M至10-13M、1×10-10M至5×10-13M、1×10-10M至1×10-12M、1×10-10M至5×10-12M、1×10-10M至1×10-11M、10-11M至10-13M、1×10-11M至5×10-13M、10- 11M至10-12M、10-12M至10-13M。在一些实施例中，抗C5a抗体与C5a之间结合的Kd值为10-7M至10-13M。In some embodiments, the anti-C5a antibody specifically binds to the C5a target with a Kd value of 10 -7 M to 10 -13 M (e.g., 10 -7 M to 10 -13 M, 10 -8 M to 10 -13 M, 10 -9 M to 10 -13 M or 10 -10 M to 10 -12 M). Therefore, in some embodiments, the Kd value of the binding between the anti-C5a antibody and C5a is 10 -7 M to 10 -13 M, 1×10 -7 M to 5×10 -13 M, 10 -7 M to 10 -12 M, 10 -7 M to 10 -11 M, 10 -7 M to 10 -10 M, 10 -7 M to 10 -9 M, 10 -8 M to 10 -13 M , 1×10 -8 M to 5×10 -13 M, 10 -8 M to 10 -12 M, 10 -8 M to 10 -11 M, 10 -8 M to 10 -10 M, 10 -8 M to 10 -9 M, 5× 10 -9 M to 1×10 -13 M, 5×10 -9 M to 1×10 -12 M, 5×10 -9 M to 1×10 -11 M, 5×10 -9 M to 1×10 -10 M, 10 -9 M to 10 -13 M, 10 -9 M to 10 -12 M, 10 -9 M to 10 -11 M, 10 -9 M to 10 -10 M, 5×10 -10 M to 1×10 -13 M, 5×10 -10 M to 1×10 - 12 M, 5×10 -10 M to 1×10 -11 M, 10 -10 M to 10 -13 M, 1×10 - 10 M to 5×10 -13 M, 1×10 -10 M to 1×10 -12 M, 1×10 -10 M to 5×10 -12 M, 1×10 -10 M to 1×10 -11 M, 10 -11 M to 10 -13 M, 1×10 -11 M to 5×10 -13 M, 10 - 11 M to 10 -12 M, 10 -12 M to 10 -13 M. In some embodiments, the Kd value for binding between the anti-C5a antibody and C5a is 10 -7 M to 10 -13 M.
在一些实施例中，抗C5a抗体与非靶标之间结合的Kd值高于抗C5a抗体与靶标的Kd值，并且本文中引用的一些实施例中，抗C5a抗体与靶标(例如，C5a)的结合亲和力高于抗C5a抗体与非靶标的结合亲和力。一些实施例中，非靶标是指非C5a的抗原。在一些实施例中，抗C5a抗体(针对C5a)与非C5a靶标结合的Kd值间至少相差10倍，例如10-100倍、100-1000倍、103-104倍、104-105倍、105-106倍、106-107倍、107-108倍、108-109倍、109-1010倍、1010-1011倍、1011-1012倍。In some embodiments, the Kd value for binding between the anti-C5a antibody and the non-target is higher than the Kd value for the anti-C5a antibody binding to the target, and in some embodiments cited herein, the Kd value for the anti-C5a antibody binding to the target (e.g., C5a) The binding affinity is higher than the binding affinity of the anti-C5a antibody to the non-target. In some embodiments, non-target refers to an antigen other than C5a. In some embodiments, the Kd value of the anti-C5a antibody (for C5a) binding to the non-C5a target is at least 10-fold different, such as 10-100 times, 100-1000 times, 10 3 -10 4 times, 10 4 -10 5 times, 10 5 -10 6 times, 10 6 -10 7 times, 10 7 -10 8 times, 10 8 -10 9 times , 10 9 -10 10 times, 10 10 -10 11 times, 10 11 -10 12 times .
在一些实施例中，所述抗C5a抗体与非靶标结合的Kd值为10-1M至10-6M(例如10-1M至10-6M、10-1M至10-5M、10-2M至10-4M)。在一些实施例中，所述非靶标是指非C5a的抗原。因此，在一些实施例中，抗C5a抗体与非C5a靶标之间结合的Kd值为10-1M至10-6M、1×10-1M至5×10-6M、10-1M至10-5M、1×10-1M至5×10-5M、10-1M至10-4M、1×10-1M至5×10-4M、10- 1M至10-3M、1×10-1M至5×10-3M、10-1M至10-2M、10-2M至10-6M、1×10-2M至5×10-6M、10-2M至10-5M、1×10-2M至5×10-5M、10-2M至10-4M、1×10-2M至5×10-4M、10-2M至10-3M、10-3M至10-6M、1×10-3M至5×10-6M、10-3M至10-5M、1×10-3M至5×10-5M、10-3M至10-4M、10-4M至10-6M、1×10-4M至5×10-6M、10-4M至10-5M、10-5M至10-6M。In some embodiments, the anti-C5a antibody binds to a non-target with a Kd value of 10 -1 M to 10 -6 M (e.g., 10 -1 M to 10 -6 M, 10 -1 M to 10 -5 M, 10 -2 M to 10 -4 M). In some embodiments, the non-target refers to an antigen other than C5a. Therefore, in some embodiments, the Kd value for binding between the anti-C5a antibody and the non-C5a target is 10 -1 M to 10 -6 M, 1×10 -1 M to 5×10 -6 M, 10 -1 M to 10 -5 M, 1×10 -1 M to 5×10 -5 M, 10 -1 M to 10 -4 M, 1× 10 -1 M to 5×10 -4 M, 10 - 1 M to 10 -3 M, 1×10 -1 M to 5×10 -3 M, 10 -1 M to 10 -2 M, 10 -2 M to 10 -6 M, 1×10 -2 M to 5×10 -6 M, 10 -2 M to 10 -5 M, 1×10 -2 M to 5×10 -5 M, 10 -2 M to 10 -4 M, 1×10 -2 M to 5×10 -4 M, 10 -2 M to 10 -3 M, 10 -3 M to 10 -6 M, 1×10 -3 M to 5×10 -6 M, 10 -3 M to 10 -5 M, 1×10 -3 M to 5×10 -5 M, 10 -3 M to 10 -4 M, 10 -4 M to 10 -6 M, 1×10 -4 M to 5×10 -6 M, 10 -4 M to 10 -5 M, 10 -5 M to 10 -6 M.
在一些实施例中，当提及抗C5a抗体以高结合亲和力特异性地识别C5a靶标，并以低结合亲和力结合非靶标时，所述抗C5a抗体与C5a靶标结合的Kd值为10-7M至10-13M(例如10-7M至10-13M、10-8M至10-13M、10-9M至10-13M、10-10M至10-12M)，并且与非靶标结合的Kd值为10-1M至10-6M(例如10-1M至10-6M、10-1M至10-5M、10-2M至10-4M)。In some embodiments, when it is referred to that an anti-C5a antibody specifically recognizes a C5a target with high binding affinity and binds to a non-target with low binding affinity, the anti-C5a antibody binds to the C5a target with a Kd value of 10 -7 M to 10 -13 M (for example, 10 -7 M to 10 -13 M, 10 -8 M to 10 -13 M, 10 -9 M to 10 -13 M, 10 -10 M to 10 -12 M), and with The Kd value for non-target binding is 10 -1 M to 10 -6 M (eg, 10 -1 M to 10 -6 M, 10 -1 M to 10 -5 M, 10 -2 M to 10 -4 M).
在一些实施例中，当提及抗C5a抗体特异性地识别C5a时，将所述抗C5a抗体的结合亲和力与对照抗C5a抗体(例如INab308)的结合亲和力进行比较。在一些实施例中，对照抗C5a抗体与C5a之间结合的Kd值可以是本申请所述的抗C5a抗体与C5a之间结合的Kd值的至少2倍，例如2倍、3倍、4倍、5倍、6倍、7倍、8倍、9倍、10倍、10-100倍、100-1000倍、103-104倍。In some embodiments, when an anti-C5a antibody is referred to as specifically recognizing C5a, the binding affinity of the anti-C5a antibody is compared to the binding affinity of a control anti-C5a antibody (eg, INab308). In some embodiments, the Kd value of the binding between the control anti-C5a antibody and C5a can be at least 2 times, such as 2 times, 3 times, 4 times the Kd value of the binding between the anti-C5a antibody and C5a described herein. , 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 10-100 times, 100-1000 times, 10 3 -10 4 times.
核酸nucleic acid
编码抗C5a抗体的核酸分子也被考虑在内。在一些实施例中，提供一种(或一组)编码全长抗C5a抗体的核酸，包括本文所述的任一种全长抗C5a抗体。在一些实施例中，本文所述的抗C5a抗体的核酸(或一组核酸)还可以包括编码多肽标签的核酸序列(例如蛋白纯化标签，His标签、HA标签)。Nucleic acid molecules encoding anti-C5a antibodies are also considered. In some embodiments, a nucleic acid (or set of) nucleic acids encoding a full-length anti-C5a antibody is provided, including any of the full-length anti-C5a antibodies described herein. In some embodiments, the nucleic acid (or set of nucleic acids) of the anti-C5a antibody described herein may also include a nucleic acid sequence encoding a polypeptide tag (eg, protein purification tag, His tag, HA tag).
同时本文还考虑了包含抗C5a抗体的分离的宿主细胞，编码抗C5a抗体多肽组分的分离的核酸，或者包含编码本文所述的抗C5a抗体多肽组分的核酸的载体。Also contemplated herein are isolated host cells comprising an anti-C5a antibody, an isolated nucleic acid encoding an anti-C5a antibody polypeptide component, or a vector comprising a nucleic acid encoding an anti-C5a antibody polypeptide component described herein.
本申请还包括这些核酸序列的变体。例如，变体包括至少在中等严格杂交条件下与编码本申请的抗C5a抗体的核酸序列杂交的核苷酸序列。Variants of these nucleic acid sequences are also encompassed by the present application. For example, variants include nucleotide sequences that hybridize under at least moderately stringent hybridization conditions to a nucleic acid sequence encoding an anti-C5a antibody of the present application.
本申请同时还提供可将本申请中核酸序列插入到其中的载体。The present application also provides a vector into which the nucleic acid sequence of the present application can be inserted.
简言之，将编码抗C5a抗体的天然或合成的核酸插入到合适的表达载体中，使得核酸可操作性的连接到5’和3’端调控元件，例如包括启动子(例如淋巴细胞特异性启动子)和3’非翻译区(UTR)，可表达抗C5a抗体(例如全长的抗C5a抗体)。所述载体可适用于在真核宿主细胞中复制和整合。典型的克隆与表达载体包含调控目标核酸序列的表达的转录和翻译终止子、起始序列和启动子。Briefly, a natural or synthetic nucleic acid encoding an anti-C5a antibody is inserted into a suitable expression vector such that the nucleic acid is operably linked to 5' and 3' regulatory elements, such as a promoter (e.g., lymphocyte-specific promoter) and 3' untranslated region (UTR), which can express anti-C5a antibodies (such as full-length anti-C5a antibodies). The vector may be suitable for replication and integration in eukaryotic host cells. Typical cloning and expression vectors contain transcriptional and translational terminators, initiation sequences, and promoters that regulate expression of the nucleic acid sequence of interest.
本申请所述的核酸也可以通过使用标准的基因递送方案，用于核酸免疫和基因治疗。核酸递送方法是本领域已知的。例如参见U.S.Pat.Nos.5,399,346、5,580,859、5,589,466，通过引用其全部内容并入本文。在一些实施例中，本申请还提供基因治疗载体。The nucleic acids described herein can also be used for nucleic acid immunization and gene therapy using standard gene delivery protocols. Nucleic acid delivery methods are known in the art. See, for example, U.S. Pat. Nos. 5,399,346, 5,580,859, 5,589,466, the entire contents of which are incorporated herein by reference. In some embodiments, the application also provides gene therapy vectors.
可以将核酸克隆到许多类型的载体中。例如，可以将核酸克隆到载体中，所述载体包括，但不限于，质粒、噬菌粒、噬菌体衍生物、动物病毒和柯斯质粒。特别感兴趣的载体包括表达载体、复制载体、探针生成载体和测序载体。Nucleic acids can be cloned into many types of vectors. For example, nucleic acids can be cloned into vectors including, but not limited to, plasmids, phagemids, phage derivatives, animal viruses, and cosmids. Vectors of particular interest include expression vectors, replication vectors, probe generation vectors and sequencing vectors.
此外，表达载体可以以病毒载体的形式提供给细胞。病毒载体技术是本领域熟知的，并且描述于例如Green and Sambrook(2013,Molecular Cloning:A Laboratory Manual,Cold Spring Harbor Laboratory,New York)，以及其它病毒学或分子生物学手册中。可用作载体的病毒包括，但不限于，逆转录病毒、腺病毒、腺相关病毒、疱疹病毒和慢病毒。通常，合适的载体包括一个在至少一种生物体中起作用的复制起点、启动子序列、方便的限制性内切酶位点以及一个或多个选择标记物(参见例如，WO 01/96584；WO 01/29058；和U.S.Pat.No.6,326,193)。Additionally, expression vectors can be provided to cells in the form of viral vectors. Viral vector technology is well known in the art and is described, for example, in Green and Sambrook (2013, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York), and other virology or molecular biology manuals. Viruses that can be used as vectors include, but are not limited to, retroviruses, adenoviruses, adeno-associated viruses, herpesviruses, and lentiviruses. Typically, suitable vectors include an origin of replication functional in at least one organism, promoter sequences, convenient restriction enzyme sites, and one or more selectable markers (see, e.g., WO 01/96584; WO 01/29058; and U.S. Pat. No. 6,326,193).
已经开发了许多基于病毒的系统，用于将基因转移到哺乳动物细胞中。例如，逆转录病毒为基因递送系统提供了便利的平台。可以应用本领域已知的技术，将选择的基因插入载体中并包装在逆转录病毒颗粒中。然后分离重组病毒，在体内或体外递送至受试者的细胞中。许多逆转录病毒系统在本领域中是已知的。在一些实施例中，使用腺病毒载体。许多腺病毒载体在本领域中是已知的。在一些实施例中，使用慢病毒载体。衍生自逆转录病毒的载体，例如慢病毒，是实现长期基因转移的合适工具，因为它们使得转基因长期稳定的整合以及在子代细胞中繁殖。慢病毒载体相对于衍生自肿瘤的逆转录病毒例如小鼠白血病病毒具有额外的优势，因为它们可以转导非分裂细胞，例如肝细胞。同时，其还具有低免疫原性的额外优势。A number of virus-based systems have been developed for gene transfer into mammalian cells. For example, retroviruses provide a convenient platform for gene delivery systems. The selected genes can be inserted into vectors and packaged in retroviral particles using techniques known in the art. The recombinant virus is then isolated and delivered to the subject's cells in vivo or in vitro. Many retroviral systems are known in the art. In some embodiments, adenoviral vectors are used. Many adenoviral vectors are known in the art. In some embodiments, lentiviral vectors are used. Vectors derived from retroviruses, such as lentiviruses, are suitable tools for long-term gene transfer because they enable long-term stable integration of the transgene and propagation in progeny cells. Lentiviral vectors have an additional advantage over tumor-derived retroviruses such as murine leukemia virus in that they can transduce non-dividing cells such as hepatocytes. At the same time, it has the additional advantage of low immunogenicity.
其它的启动子元件，例如，增强子，调控转录起始频率。通常它们位于起始位点上游30-110bp处，虽然最近发现很多启动子也包含起始位点下游的功能元件。启动子元件之间的间隔通常是灵活的，所以当元件彼此之间位置互换或移动时仍保持启动子的功能。在胸苷激酶(tk)启动子中，启动子元件之间的间隔增加到50bp活性才会开始下降。Other promoter elements, such as enhancers, regulate the frequency of transcription initiation. Typically they are located 30-110 bp upstream of the start site, although recently many promoters have been found to also contain functional elements downstream of the start site. The spacing between promoter elements is usually flexible, so that promoter function is maintained when elements are interchanged or moved with each other. In the thymidine kinase (tk) promoter, activity does not begin to decrease until the spacing between promoter elements increases to 50 bp.
合适启动子的一个示例是即时早期巨细胞病毒(CMV)启动子序列。该启动子序列是一个很强的组成型启动子序列，可以驱动任何与其可操作性连接的多核苷酸序列高水平表达。合适启动子的另一个示例是延伸因子1α(EF-1α)启动子。然而，也可以使用其它组成型启动子，包括但不限于猿猴病毒40(SV40)早期启动子、小鼠乳腺肿瘤病毒(MMTV)、人免病缺陷病毒长末端重复序列(HIV-LTR)启动子、MoMuLV启动子、禽类白血病病毒启动子、Epstein-Barr病毒即刻早期启动子、劳斯氏肉瘤病毒启动子以及人类基因启动子，例如包括但不限于肌动蛋白启动子、肌球蛋白启动子、血红蛋白启动子和肌酸激酶启动子。此外，不应将本申请局限在仅使用组成型启动子，诱导型启动子也是本申请考虑的部分。诱导型启动子的使用提供了一种分子开关，当需要这种表达时，能启动其与之可操作性连接的多核苷酸序列表达，当不需要时，则关闭表达。诱导型启动子包含，但不局限于，金属硫蛋白启动子、糖皮质激素启动子、孕酮启动子和四环素启动子。An example of a suitable promoter is the immediate early cytomegalovirus (CMV) promoter sequence. This promoter sequence is a strong constitutive promoter sequence that can drive high-level expression of any polynucleotide sequence operably linked to it. Another example of a suitable promoter is the elongation factor 1α (EF-1α) promoter. However, other constitutive promoters may also be used, including, but not limited to, simian virus 40 (SV40) early promoter, mouse mammary tumor virus (MMTV), human immunodeficiency virus long terminal repeat (HIV-LTR) promoter , MoMuLV promoter, avian leukemia virus promoter, Epstein-Barr virus immediate early promoter, Rous sarcoma virus promoter and human gene promoters, such as but not limited to actin promoter, myosin promoter, Hemoglobin promoter and creatine kinase promoter. Furthermore, this application should not be limited to the use of only constitutive promoters, inducible promoters are also considered as part of this application. The use of an inducible promoter provides a molecular switch that turns on the expression of the polynucleotide sequence to which it is operably linked when such expression is desired, and turns off expression when it is not. Inducible promoters include, but are not limited to, metallothionein promoters, glucocorticoid promoters, progesterone promoters, and tetracycline promoters.
在一些实施例中，抗C5a抗体的表达是可诱导的。在一些实施例中，编码抗C5a抗体的核酸序列可操作的连接到诱导型启动子上，包括本文所述的任一诱导型启动子。In some embodiments, expression of anti-C5a antibodies is inducible. In some embodiments, a nucleic acid sequence encoding an anti-C5a antibody is operably linked to an inducible promoter, including any of the inducible promoters described herein.
诱导型启动子inducible promoter
诱导型启动子的使用提供了一种分子开关，当需要表达时，可启动与之可操作性连接的多核苷酸序列表达，而在不需要表达时，则关闭表达。真核细胞中适用的示例性诱导型启动子包括，但不限于，激素调节元件(例如，参见Mader,S.andWhite,J.H.(1993)Proc.Natl.Acad.Sci.USA 90:5603-5607)、合成配体调节元件(参见Spencer,D.M.et al(1993)Science 262:1019-1024)以及电离辐射调控元件(参见Manome,Y.et al.(1993)Biochemistry 32:10607-10613；Datta,R.et al.(1992)Proc.Natl.Acad.Sci.USA 89:1014-10153)。其它适用于体内或体外哺乳动物系统的示例性诱导型启动子参见Gingrich et al.(1998)Annual Rev.Neurosci 21:377-405。在一些实施例中，用于表达抗C5a抗体的诱导型启动子系统为Tet系统。在一些实施例中，用于表达抗C5a抗体的诱导型启动子系统为大肠杆菌lac抑制系统。The use of an inducible promoter provides a molecular switch that turns on the expression of a polynucleotide sequence operably linked to it when expression is desired, and turns off expression when expression is not needed. Exemplary inducible promoters suitable for use in eukaryotic cells include, but are not limited to, hormone regulatory elements (see, e.g., Mader, S. and White, J.H. (1993) Proc. Natl. Acad. Sci. USA 90:5603-5607) , synthetic ligand regulatory elements (see Spencer, D.M. et al (1993) Science 262:1019-1024) and ionizing radiation regulatory elements (see Manome, Y. et al. (1993) Biochemistry 32:10607-10613; Datta, R .et al.(1992)Proc.Natl.Acad.Sci.USA 89:1014-10153). Other exemplary inducible promoters suitable for use in mammalian systems in vivo or in vitro are described in Gingrich et al. (1998) Annual Rev. Neurosci 21:377-405. In some embodiments, the inducible promoter system used to express anti-C5a antibodies is the Tet system. In some embodiments, the inducible promoter system used to express anti-C5a antibodies is an E. coli lac repressor system.
本申请所采用的一个示例性诱导型启动子系统为Tet系统。该系统是基于Gossen等(1993)描述的Tet系统。在一个示例性实施例中，目标多核苷酸由包含一个或多个Tet操纵子(TetO)位点的启动子控制。在非激活状态，Tet阻遏物(TetR)与TetO位点结合并抑制启动子的转录。在激活状态，例如，在存在诱导剂如四环素(Tc)、无水四环素、多西环素(Dox)或其活性类似物的情况下，诱导剂会使TetR从TetO上释放，从而导致转录发生。多西环素是四环素抗生素家族中的一员，其化学名为1-二甲氨基-2,4a,5,7-五羟基-11-甲基-4,6-二氧基-1,4a,11,11a,12,12a-六氢四烯-3-甲酰胺。An exemplary inducible promoter system used in this application is the Tet system. This system is based on the Tet system described by Gossen et al. (1993). In an exemplary embodiment, the target polynucleotide is controlled by a promoter containing one or more Tet operator (TetO) sites. In the inactive state, Tet repressor (TetR) binds to the TetO site and inhibits transcription from the promoter. In the activated state, for example, in the presence of inducers such as tetracycline (Tc), anhydrotetracycline, doxycycline (Dox) or their active analogs, the inducer causes TetR to be released from TetO, resulting in transcription. . Doxycycline is a member of the tetracycline antibiotic family, its chemical name is 1-dimethylamino-2,4a,5,7-pentahydroxy-11-methyl-4,6-dioxy-1,4a ,11,11a,12,12a-hexahydrotetraene-3-carboxamide.
在一个实施例中，TetR经密码子优化适用于在哺乳动物细胞中表达，例如小鼠或人类细胞。由于遗传密码的简并性，大多数氨基酸由不止一个密码子编码，从而使得给定核酸的序列具有大量的变体，而其编码的氨基酸序列没有任何改变。然而，许多生物体在密码子使用方面存在差异，也称为“密码子偏好”(即，给定氨基酸使用特定密码子的偏好)。密码子偏好通常与特定密码子的优势tRNA种类的存在有关，反过来又提高了mRNA翻译的效率。因此可以通过密码子优化来定制源自特定物种的编码序列(例如，原核生物)，以提高其在不同物种(例如，真核生物)中的表达。In one embodiment, TetR is codon-optimized for expression in mammalian cells, such as mouse or human cells. Due to the degeneracy of the genetic code, most amino acids are encoded by more than one codon, resulting in a large number of variants in the sequence of a given nucleic acid without any change in the sequence of its encoded amino acid. However, many organisms have differences in codon usage, also known as "codon preference" (i.e., the preference for a given amino acid to use a specific codon). Codon preference is often associated with the presence of dominant tRNA species for specific codons, which in turn increases the efficiency of mRNA translation. Coding sequences derived from a specific species (e.g., prokaryotes) can thus be tailored through codon optimization to enhance their expression in different species (e.g., eukaryotes).
Tet系统的其它具体变体，包括以下的“Tet-Off”和“Tet-On”系统。在Tet-off系统中，转录在Tc或Dox存在下是失活的。在该系统中，由TetR与单纯疱疹病毒VP16强转录激活结构域融合组成的四环素调控的转录激活蛋白(tTA)，在四环素反应启动子元件(TRE)转录控制下调控靶核酸的表达。TRE元件由TetO序列串联与启动子(通常是来源于人巨细胞病毒即刻早期启动子的最小启动子序列)融合组成。在不存在Tc或Dox的情况下，tTA结合TRE并激活靶基因的转录。在存在Tc或Dox的情况下，tTA不能结合TRE，靶基因不能表达。Other specific variations of the Tet system include the "Tet-Off" and "Tet-On" systems below. In the Tet-off system, transcription is inactive in the presence of Tc or Dox. In this system, the tetracycline-regulated transcriptional activator (tTA), which is composed of the fusion of TetR and the strong transcriptional activation domain of herpes simplex virus VP16, regulates the expression of target nucleic acids under the transcriptional control of the tetracycline-responsive promoter element (TRE). The TRE element consists of a tandem TetO sequence fused to a promoter (usually a minimal promoter sequence derived from the human cytomegalovirus immediate early promoter). In the absence of Tc or Dox, tTA binds TREs and activates transcription of target genes. In the presence of Tc or Dox, tTA cannot bind to TRE and target genes cannot be expressed.
相反，在Tet-On系统中，转录在Tc或Dox存在下是激活的。Tet-On系统是基于反向四环素调控的转录激活因子rtTA。与tTA一样，rtTA是由TetR阻遏物与VP16转录激活结构域组成的融合蛋白。然而，TetR的DNA结合区中4个氨基酸的变化改变了rtTA的结合特性，使其在存在Dox的情况下只能识别靶转基因TRE上的tetO序列。所以在Tet-On系统中，只有在存在Dox的情况下，rtTA才能激活TRE调控的靶基因的转录。In contrast, in the Tet-On system, transcription is activated in the presence of Tc or Dox. The Tet-On system is based on the reverse tetracycline-regulated transcriptional activator rtTA. Like tTA, rtTA is a fusion protein composed of the TetR repressor and the VP16 transcriptional activation domain. However, a 4-amino acid change in the DNA-binding region of TetR changes the binding properties of rtTA, causing it to only recognize the tetO sequence on the target transgene TRE in the presence of Dox. Therefore, in the Tet-On system, rtTA can activate the transcription of TRE-regulated target genes only in the presence of Dox.
另一种诱导型启动子系统是大肠杆菌的lac阻遏物系统(参见Brown et al.,Cell49:603-612(1987))。Lac阻遏物系统通过调控与包含lac操纵子(lacO)的启动子可操作性连接的目标多核苷酸的转录发挥功能。Lac阻遏物(lacR)与LacO结合，进而阻止目标多核苷酸的转录。通过合适的诱导剂来诱导目标多核苷酸的表达，例如，异丙基-β-D硫代半乳糖吡喃苷(IPTG)。Another inducible promoter system is the lac repressor system of E. coli (see Brown et al., Cell 49:603-612 (1987)). The Lac repressor system functions by regulating the transcription of a target polynucleotide operably linked to a promoter containing the lac operator (lacO). Lac repressor (lacR) binds to LacO, thereby preventing the transcription of the target polynucleotide. Expression of the polynucleotide of interest is induced by a suitable inducer, for example, isopropyl-β-D thiogalactopyranoside (IPTG).
为了评估多肽或其部分的表达，待导入细胞的表达载体还可包含选择标记基因或报告基因或二者都有，以便于从病毒载体转染或感染的细胞群体中识别和选择表达细胞。在其他方面，选择标记可以携带在单独的DNA片段上并在共转染实验中使用。选择标记基因或报告基因都可侧接于合适的调控序列，使其在宿主细胞中能够表达。有用的选择标记包括，例如，抗生素耐药基因，如neo以及类似基因。To assess the expression of a polypeptide or portion thereof, the expression vector to be introduced into the cell may also contain a selectable marker gene or a reporter gene or both to facilitate the identification and selection of expressing cells from a population of cells transfected or infected with the viral vector. In other aspects, the selectable marker can be carried on separate DNA fragments and used in co-transfection experiments. Either the selectable marker gene or the reporter gene can be flanked by appropriate regulatory sequences to enable expression in the host cell. Useful selectable markers include, for example, antibiotic resistance genes such as neo and similar genes.
报告基因可用于鉴定潜在的转染细胞和评价调控序列的功能。通常，报告基因是不存在于受体生物体或组织中或不由受体生物体或组织表达的基因，其编码一种多肽，其表达表现为一些易于检测的特性，例如酶活性。当DNA导入受体细胞后，在合适的时间检测报告基因的表达。合适的报告基因可包括编码荧光素酶、β-半乳糖苷酶、氯霉素乙酰转移酶、分泌碱性磷酸酶或绿色荧光蛋白的基因(参见,Ui-Tel et al.,2000 FEBS Letters 479:79-82)。合适的表达系统是公知的，可以通过已知的技术制备或通过商业途径获得。通常，把可显示报告基因最高表达水平的最小5’侧翼区的构建体认定为启动子。此类启动子区可以与报告基因连接，并用于评估某些物质在调节启动子驱动的转录中能力。Reporter genes can be used to identify potentially transfected cells and evaluate the function of regulatory sequences. Typically, a reporter gene is a gene that is not present in or expressed by the recipient organism or tissue and encodes a polypeptide whose expression is manifested by some readily detectable property, such as enzymatic activity. After the DNA is introduced into the recipient cells, the expression of the reporter gene is detected at the appropriate time. Suitable reporter genes may include genes encoding luciferase, β-galactosidase, chloramphenicol acetyltransferase, secreted alkaline phosphatase, or green fluorescent protein (see, Ui-Tel et al., 2000 FEBS Letters 479 :79-82). Suitable expression systems are well known and can be prepared by known techniques or commercially available. Typically, the construct with the smallest 5' flanking region that shows the highest expression level of the reporter gene is considered the promoter. Such promoter regions can be linked to reporter genes and used to assess the ability of certain substances to regulate promoter-driven transcription.
在一些实施例中，提供编码本文所述的任一种全长抗C5a抗体的核酸。在一些实施例中，所述核酸包括编码全长抗C5a抗体重链和轻链的一个或多个核酸序列。在一些实施例中，所述一个或多个核酸序列中的每一个包含在单独的载体中。在一些实施例中，至少有一些核酸序列包含在同一载体中。在一些实施例中，所有核酸序列包含在同一载体中。载体可以选自，例如，哺乳动物表达载体和病毒载体(如源自逆转录病毒、腺病毒、腺相关病毒、疱疹病毒和慢病毒的载体)。In some embodiments, nucleic acids encoding any of the full-length anti-C5a antibodies described herein are provided. In some embodiments, the nucleic acid includes one or more nucleic acid sequences encoding full-length anti-C5a antibody heavy and light chains. In some embodiments, each of the one or more nucleic acid sequences is contained in a separate vector. In some embodiments, at least some of the nucleic acid sequences are included in the same vector. In some embodiments, all nucleic acid sequences are contained in the same vector. Vectors may be selected, for example, from mammalian expression vectors and viral vectors (eg, vectors derived from retroviruses, adenoviruses, adeno-associated viruses, herpesviruses and lentiviruses).
将基因导入细胞并表达的方法在本领域是已知的。在涉及表达载体的上下文中，通过本领域的任何方法载体可以很容易地导入宿主细胞中，如哺乳动物细胞、细菌、酵母或昆虫细胞。例如表达载体可以通过物理、化学或生物方法导入宿主细胞。Methods of introducing genes into cells and expressing them are known in the art. In the context of expression vectors, the vector can be readily introduced into host cells, such as mammalian cells, bacteria, yeast or insect cells, by any method in the art. For example, expression vectors can be introduced into host cells by physical, chemical or biological methods.
将多核苷酸导入宿主细胞的物理方法包括磷酸钙沉淀、脂质体转染、基因枪法、显微注射、电穿孔法以及诸如此类。制备包含载体和/或外源核酸的细胞的方法在本领域是熟知的。参见例如Green and Sambrook(2013,Molecular Cloning:A Laboratory Manual,Cold Spring Harbor Laboratory,New York)。在一些实施例中，通过磷酸钙转染法将多核苷酸导入宿主细胞。Physical methods of introducing polynucleotides into host cells include calcium phosphate precipitation, lipofection, biolistic methods, microinjection, electroporation, and the like. Methods of preparing cells containing vectors and/or exogenous nucleic acids are well known in the art. See, for example, Green and Sambrook (2013, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, New York). In some embodiments, the polynucleotide is introduced into the host cell by calcium phosphate transfection.
将目标多核苷酸导入宿主细胞的生物学方法包括使用DNA和RNA载体。病毒载体，特别是逆转录病毒载体，已成为将基因插入哺乳动物细胞，例如人类细胞中的最广泛使用的方法。其它病毒载体可以源自慢病毒、痘病毒、单纯疱疹病毒1型、腺病毒和腺相关病毒等。参见如U.S.Pat.Nos.5,350,674和5,585,362。Biological methods for introducing polynucleotides of interest into host cells include the use of DNA and RNA vectors. Viral vectors, especially retroviral vectors, have become the most widely used method for inserting genes into mammalian cells, such as human cells. Other viral vectors can be derived from lentivirus, poxvirus, herpes simplex virus type 1, adenovirus, adeno-associated virus, etc. See, for example, U.S. Pat. Nos. 5,350,674 and 5,585,362.
将多核苷酸导入宿主细胞的化学方法包括胶体分散系统，例如高分子复合物、纳米胶囊、微球、磁珠和以脂质为基础的系统，其包括水包油乳剂、胶团、混合胶团和脂质体。一种在体内和体外被用作递送载体的示例性胶体系统是脂质体(例如，人工膜囊)。Chemical methods for introducing polynucleotides into host cells include colloidal dispersion systems such as polymer complexes, nanocapsules, microspheres, magnetic beads, and lipid-based systems including oil-in-water emulsions, micelles, and mixed gels pellets and liposomes. One exemplary colloidal system used as a delivery vehicle both in vivo and in vitro is liposomes (eg, artificial membrane vesicles).
在使用非病毒递送系统的情况下，示例性的递送载体是脂质体。考虑使用脂质制剂将核酸导入宿主细胞(体外、离体或体内)。在另一方面，所述核酸可以与脂质结合。与脂质结合的核酸可被包裹进脂质体的水性内部，散布在脂质体的脂质双层内，通过与脂质体和寡核苷酸结合的连接分子连接在脂质体，包埋在脂质体中，与脂质体形成复合物，分散在含有脂质的溶液中，与脂质混合，与脂质结合，悬浮在脂质中，包含在胶束中或与胶束混合，或以其它方式与脂质结合。脂质、脂质/DNA或脂质/表达载体相关的组合物在溶液中不限于任何特定结构。例如，它们可能以双分子层结构、以胶束或以“塌陷”结构存在。它们也可以简单的分散在溶液中，可能形成大小或形状不均匀的聚集体。脂质是脂肪物质，可以是天然存在的或是合成的脂质。例如，脂质包括天然存在于细胞质中的脂肪滴，以及含有长链脂肪烃及其衍生物的一类化合物，例如脂肪酸、醇、胺、氨基醇和醛。Where non-viral delivery systems are used, an exemplary delivery vehicle is liposomes. Consider using lipid formulations to introduce nucleic acids into host cells (in vitro, ex vivo, or in vivo). In another aspect, the nucleic acid can be associated with lipids. The nucleic acid bound to the lipid can be packaged into the aqueous interior of the liposome, spread within the lipid bilayer of the liposome, and connected to the liposome through a linker molecule that binds to the liposome and the oligonucleotide. Buried in liposomes, forming complexes with liposomes, dispersed in solutions containing lipids, mixed with lipids, bound to lipids, suspended in lipids, contained in micelles or mixed with micelles , or otherwise combined with lipids. Lipid, lipid/DNA or lipid/expression vector related compositions are not limited to any particular structure in solution. For example, they may exist in bilayer structures, in micelles or in "collapsed" structures. They can also simply disperse in solution, possibly forming aggregates that are not uniform in size or shape. Lipids are fatty substances that can be naturally occurring or synthetic lipids. For example, lipids include lipid droplets that occur naturally in the cytoplasm, as well as a class of compounds containing long-chain aliphatic hydrocarbons and their derivatives, such as fatty acids, alcohols, amines, aminoalcohols, and aldehydes.
无论采用何种方法将外源核酸导入宿主细胞中或以其他方式将细胞暴露于本申请的抑制剂中，为了确认重组DNA序列存在于宿主细胞中，可以进行多种实验。这类实验包括例如本领域技术人员熟知的“分子生物学”实验。例如Southern和Northern blotting，RT-PCR和PCR；“生物化学”实验，例如检测某一特定多肽存在或不存在，例如通过免疫学方法(ELISAs和Western blots)或者通过本文所述的实验来进行鉴定均落入本申请范围内。Regardless of which method is used to introduce exogenous nucleic acid into a host cell or otherwise expose the cell to the inhibitor of the present application, a variety of experiments can be performed in order to confirm that the recombinant DNA sequence is present in the host cell. Such experiments include, for example, "molecular biology" experiments well known to those skilled in the art. For example, Southern and Northern blotting, RT-PCR and PCR; "biochemical" experiments, such as detecting the presence or absence of a specific polypeptide, such as by immunological methods (ELISAs and Western blots) or by experiments described in this article All fall within the scope of this application.
抗C5a抗体的制备Preparation of anti-C5a antibodies
在一些实施例中，所述抗C5a抗体是单克隆抗体或源于单克隆抗体。在一些实施例中，所述抗C5a抗体包含来自单克隆抗体的VH和VL，或者其变体。在一些实施例中，所述抗C5a抗体进一步包括来自单克隆抗体的CH1和CL区域，或者其变体。单克隆抗体可以应用例如本领域已知的方法制备，包括杂交瘤细胞法、噬菌体展示方法或应用重组DNA法。此外，示例性的噬菌体展示法在本文及以下的实施例中进行了描述。In some embodiments, the anti-C5a antibody is a monoclonal antibody or is derived from a monoclonal antibody. In some embodiments, the anti-C5a antibody comprises VH and VL from a monoclonal antibody, or a variant thereof. In some embodiments, the anti-C5a antibody further includes CH1 and CL regions from a monoclonal antibody, or variants thereof. Monoclonal antibodies can be prepared using, for example, methods known in the art, including hybridoma cell methods, phage display methods, or using recombinant DNA methods. Additionally, exemplary phage display methods are described herein and in the Examples below.
在杂交瘤细胞法中，通常采用免疫剂免疫仓鼠、小鼠或其他适合的宿主动物，以引发产生或能够产生与免疫剂特异性结合的抗体的淋巴细胞。或者，可以在体外免疫淋巴细胞。免疫剂可包括目标蛋白的多肽或融合蛋白。通常，如果需要人源细胞，采用外周血淋巴细胞(PBLs)，而如果需要非人哺乳动物来源细胞，则会使用脾细胞或淋巴结细胞。使用适当的融合剂将淋巴细胞与永生细胞系进行融合，例如聚乙二醇，以形成杂交瘤细胞。永生细胞系通常是转化的哺乳动物细胞，尤其是啮齿类、牛科和人源的骨髓瘤细胞。通常采用大鼠或小鼠骨髓瘤细胞系。杂交瘤细胞可以在合适的培养基中进行培养，所述培养基优选含有一种或多种抑制未融合永生细胞生长或存活的物质。例如，如果亲本细胞缺乏次黄嘌呤-鸟嘌呤磷酸核糖转移酶(HGPRT或HPRT)，则杂交瘤细胞培养基通常包括次黄嘌呤、氨蝶呤和胸苷(HAT培养基)，该培养基能阻止HGPRT缺陷细胞生长。In the hybridoma cell method, an immune agent is usually used to immunize hamsters, mice or other suitable host animals to induce lymphocytes that produce or are capable of producing antibodies that specifically bind to the immune agent. Alternatively, lymphocytes can be immunized in vitro. Immunizing agents may include polypeptides or fusion proteins of the protein of interest. Typically, if cells of human origin are desired, peripheral blood lymphocytes (PBLs) are used, whereas if cells of non-human mammalian origin are desired, spleen cells or lymph node cells are used. Lymphocytes are fused to immortalized cell lines using an appropriate fusion agent, such as polyethylene glycol, to form hybridoma cells. Immortal cell lines are typically transformed mammalian cells, particularly myeloma cells of rodent, bovine, and human origin. Typically, rat or mouse myeloma cell lines are used. Hybridoma cells can be cultured in a suitable medium, which preferably contains one or more substances that inhibit the growth or survival of unfused immortal cells. For example, if the parent cells lack the enzyme hypoxanthine-guanine phosphoribosyltransferase (HGPRT or HPRT), the hybridoma cell culture medium usually includes hypoxanthine, aminopterin, and thymidine (HAT medium), which can Prevents the growth of HGPRT-deficient cells.
在一些实施例中，永生化细胞系有效融合，通过所选择的抗体生产细胞保证抗体高水平稳定表达，并且对某些培养基敏感，例如HAT培养基。在一些实施例中，永生细胞系是小鼠骨髓瘤细胞系，可以从例如，加利福尼亚圣地亚哥的索尔克细胞保藏中心和弗吉尼亚马纳萨斯的美国典型培养物保藏中心获得。同时还描述了人骨髓瘤和鼠-人杂交骨髓瘤细胞系用于制备人源单克隆抗体。In some embodiments, the immortalized cell lines fuse efficiently, ensure high-level and stable expression of the antibody by the selected antibody-producing cells, and are sensitive to certain media, such as HAT media. In some embodiments, the immortal cell line is a mouse myeloma cell line, available from, for example, the Salk Cell Collection in San Diego, California, and the American Type Culture Collection in Manassas, Virginia. Also described are human myeloma and mouse-human hybrid myeloma cell lines for the preparation of human monoclonal antibodies.
然后可以测定培养杂交瘤细胞的培养基中是否存在针对多肽的单克隆抗体。由杂交瘤细胞产生的单克隆抗体的结合特异性可以通过免疫沉淀法或体外结合实验确定，如放射性免疫测定法(RIA)或酶联免疫吸附法(ELISA)。此类技术或分析方法在本领域是已知的。单克隆抗体的结合亲和力可以通过例如Munson and Pollard,Anal.Biochem.,107:220(1980)中所述的斯卡查德(Scatchard)分析确定。The culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against the polypeptide. The binding specificity of monoclonal antibodies produced by hybridoma cells can be determined by immunoprecipitation or in vitro binding experiments, such as radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). Such techniques or analytical methods are known in the art. The binding affinity of a monoclonal antibody can be determined by Scatchard analysis as described, for example, in Munson and Pollard, Anal. Biochem., 107:220 (1980).
在鉴定出所需的杂交瘤细胞后，可以通过有限稀释法对目标克隆进行亚克隆，并通过标准方法进行培养。基于此目的适合的培养基包括，例如改良Eagle培养基(DMEM)和RPMI-1640培养基。或者，杂交瘤细胞可以在哺乳动物体内以腹水的形式生长。After the desired hybridoma cells are identified, the clones of interest can be subcloned by limiting dilution and cultured by standard methods. Suitable media for this purpose include, for example, modified Eagle's medium (DMEM) and RPMI-1640 medium. Alternatively, hybridoma cells can be grown in mammals in the form of ascites fluid.
亚克隆分泌的单克隆抗体可以通过常规免疫球蛋白纯化方法从培养基或腹水中分离或纯化，例如蛋白A-琼脂糖凝胶、羟基磷灰石色谱层析、凝胶电泳、透析或亲和层析。Monoclonal antibodies secreted by the subclones can be isolated or purified from the culture medium or ascitic fluid by conventional immunoglobulin purification methods, such as protein A-Sepharose, hydroxyapatite chromatography, gel electrophoresis, dialysis, or affinity Chromatography.
在一些实施例中，根据本文所述的任一抗C5a抗体，所述抗C5a抗体包含选自抗体文库(例如展示scFv或Fab片段的噬菌体文库)的克隆的序列。所述克隆可以通过筛选具有所需活性的抗体片段组合文库的方法进行鉴定。例如，本领域已知多种方法用于产生噬菌体展示文库以及筛选这些文库来获得所需结合特性的抗体。这些方法在例如Hoogenboom et al.,Methods in Molecular Biology 178:1-37(O'Brien et al.,ed.,Human Press,Totowa,N.J.,2001)中进行了综述，并且在例如McCafferty et al.,Nature 348:552-554；Clackson et al.,Nature 352:624-628(1991)；Marks et al.,J.Mol.Biol.222:581-597(1992)；Marks and Bradbury,Methods in Molecular Biology 248:161-175(Lo,ed.,Human Press,Totowa,N.J.,2003)；Sidhu et al.,J.Mol.Biol.338(2):299-310(2004)；Lee et al.,J.Mol.Biol.340(5):1073-1093(2004)；Fellouse,Proc.Natl.Acad.Sci.USA 101(34):12467-12472(2004)；and Lee et al.,J.Immunol.Methods 284(1-2):119-132(2004)中进行了进一步描述。In some embodiments, according to any anti-C5a antibody described herein, the anti-C5a antibody comprises sequences selected from clones of an antibody library (eg, a phage library displaying scFv or Fab fragments). The clones can be identified by screening a combinatorial library of antibody fragments with the desired activity. For example, various methods are known in the art for generating phage display libraries and screening these libraries for antibodies with desired binding properties. These methods are reviewed, for example, in Hoogenboom et al., Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press, Totowa, N.J., 2001), and in, e.g., McCafferty et al. ,Nature 348:552-554;Clackson et al.,Nature 352:624-628(1991);Marks et al.,J.Mol.Biol.222:581-597(1992);Marks and Bradbury,Methods in Molecular Biology 248:161-175(Lo,ed.,Human Press,Totowa,N.J.,2003);Sidhu et al.,J.Mol.Biol.338(2):299-310(2004);Lee et al., J.Mol.Biol.340(5):1073-1093(2004); Fellouse, Proc.Natl.Acad.Sci.USA 101(34):12467-12472(2004); and Lee et al., J.Immunol .Methods 284(1-2):119-132(2004) is further described.
在某些噬菌体展示方法中，通过聚合酶链式反应(PCR)分别克隆VH和VL基因的所有组成成分，并在噬菌体文库中随机重组，然后筛选能够结合抗原的噬菌体，如Winter et al.,Ann.Rev.Immunol.,12:433-455(1994)中所述。噬菌体通常以scFv片段或以Fab片段形式展示抗体片段。免疫来源的文库噬菌体提供针对免疫原的高亲和力抗体而不需要构建杂交瘤细胞。或者，可以克隆天然库(例如来自人)，来提供针对多种非自身抗原和自身抗原的单一抗体来源，而不需任何免疫，如Griffiths et al.,EMBOJ,12:725-734(1993)中所述。最后，天然文库也可以通过克隆来自干细胞的非重排V-gene片段，并使用包含随机序列的PCR引物编码CDR3高变区并且在体外完成重排的方法进行制备，如Hoogenboom and Winter,J.Mol.Biol.,227:381-388(1992)中所述。描述人抗体噬菌体文库的专利出版物包括，例如U.S.Pat.No.5,750,373和US Patent Publication Nos.2005/0079574、2005/0119455、2005/0266000、2007/0117126、2007/0160598、2007/0237764、2007/0292936和2009/0002360。In some phage display methods, all components of the V H and V L genes are cloned separately through polymerase chain reaction (PCR), randomly recombined in a phage library, and then screened for phages that can bind the antigen, such as Winter et al. ., Ann. Rev. Immunol., 12:433-455 (1994). Phages typically display antibody fragments as scFv fragments or as Fab fragments. Immunogenically derived library phages provide high-affinity antibodies against immunogens without the need to construct hybridoma cells. Alternatively, natural libraries (eg from humans) can be cloned to provide a single source of antibodies against multiple non-self and self-antigens without the need for any immunization, as in Griffiths et al., EMBOJ, 12:725-734 (1993) described in. Finally, natural libraries can also be prepared by cloning non-rearranged V-gene fragments from stem cells and using PCR primers containing random sequences to encode the CDR3 hypervariable region and completing the rearrangement in vitro, such as Hoogenboom and Winter, J. Mol. Biol., 227:381-388 (1992). Patent publications describing human antibody phage libraries include, for example, US Pat. No. 5,750,373 and US Patent Publication Nos. 2005/0079574, 2005/0119455, 2005/0266000, 2007/0117126, 2007/0160598, 2007/0237764, 2007/0 292936 and 2009/0002360.
通过噬菌体展示筛选文库中能够特异性结合靶标C5a的抗C5a抗体部分的方法来制备所述的抗C5a抗体。该文库可以是人scFv噬菌体展示文库，具有至少1×109(例如至少1×109、2.5×109、5×109、7.5×109、1×1010、2.5×1010、5×1010、7.5×1010或1×1011)种多样性的独特的人抗体片段。在一些实施例中，所述文库是人天然文库，通过从健康受试者的PMBCs和脾脏中提取的DNA构建，包含所有人重链和轻链亚家族。在一些实施例中，所述文库是人天然文库，通过从各种疾病患者体内分离的PMBCs中提取的DNA构建，例如自身免疫病的患者、癌症患者和感染性疾病的患者。在一些实施例中，所述文库是半合成的人文库，其中重链CDR3完全是随机的，所有氨基酸(除了半胱氨酸)以相同的概率存在于任何给定的位置。(参见例如,Hoet,R.M.et al.,Nat.Biotechnol.23(3):344-348,2005)。在一些实施例中，半合成的人文库的重链CDR3长度在5到24个(例如5、6、7、8、9、10、11、12、13、14、15、16、17、18、19、20、21、22、23或24个)氨基酸之间。在一些实施例中，所述文库是全合成的噬菌体展示文库。在一些实施例中，所述文库是非人噬菌体展示文库。The anti-C5a antibody is prepared by phage display screening library for anti-C5a antibody portions that can specifically bind to target C5a. The library may be a human scFv phage display library with at least 1×10 9 (e.g., at least 1×10 9 , 2.5×10 9 , 5×10 9 , 7.5×10 9 , 1×10 10 , 2.5×10 10 , 5 ×10 10 , 7.5×10 10 or 1×10 11 ) diversity of unique human antibody fragments. In some embodiments, the library is a human natural library, constructed from DNA extracted from PMBCs and spleens of healthy subjects, containing all human heavy and light chain subfamilies. In some embodiments, the library is a human natural library constructed from DNA extracted from PMBCs isolated from patients with various diseases, such as patients with autoimmune diseases, cancer patients, and patients with infectious diseases. In some embodiments, the library is a semi-synthetic human library in which the heavy chain CDR3s are completely random and all amino acids (except cysteine) are present with equal probability at any given position. (See, eg, Hoet, RM et al., Nat. Biotechnol. 23(3):344-348, 2005). In some embodiments, the heavy chain CDR3 length of the semi-synthetic human library ranges from 5 to 24 (e.g., 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18 , 19, 20, 21, 22, 23 or 24) between amino acids. In some embodiments, the library is a fully synthetic phage display library. In some embodiments, the library is a non-human phage display library.
与靶标C5a具有高亲和力的噬菌体克隆可以通过噬菌体与靶标C5a的迭代结合进行筛选，所述靶标C5a与固相支持物结合(例如用于溶液淘选的珠子或用于细胞淘选的哺乳动物细胞)，接下来去除未结合的噬菌体，并洗脱特异性结合噬菌体。随后，洗脱结合的噬菌体克隆并用于感染合适的宿主细胞，例如E.coli XL1-Blue，进行表达和纯化。可以通过多轮淘选(例如，2、3、4、5、6或更多轮)，例如溶液淘选、细胞淘选或两者结合以富集特异性结合C5a的噬菌体克隆。富集的噬菌体克隆与靶标C5a的特异性结合可以通过本领域已知的任何方法进行检测，包括例如ELISA和FACS。Phage clones with high affinity for target C5a can be screened by iterative binding of phage to target C5a bound to a solid support (e.g., beads for solution panning or mammalian cells for cell panning) ), followed by removal of unbound phage and elution of specifically bound phage. Subsequently, bound phage clones are eluted and used to infect suitable host cells, such as E. coli XL1-Blue, for expression and purification. Phage clones that specifically bind C5a can be enriched by multiple rounds of panning (eg, 2, 3, 4, 5, 6, or more rounds), such as solution panning, cell panning, or a combination of both. Specific binding of the enriched phage clones to the target C5a can be detected by any method known in the art, including, for example, ELISA and FACS.
筛选抗体文库的另一种方法是在酵母细胞表面展示蛋白质。Wittrup等(美国专利6,699,658和6,696,251)开发了一种酵母细胞展示文库的方法。在此酵母展示系统中，一个组分包括锚定在酵母细胞壁上的酵母凝集素蛋白(Aga1)，另一个组分包括凝集素蛋白Aga2的第二个亚基，该亚基可以通过二硫键与Aga1蛋白结合进而展示在酵母细胞表面上。通过将Aga1基因整合到酵母染色体中来表达Aga1蛋白。将单链可变片段(scFv)文库与酵母展示质粒中的Aga2基因融合，将其转化后，该文库由于附加的营养标记的存在可保留在酵母中。Aga1和Aga2蛋白均在半乳糖诱导型启动子的控制下表达。Another way to screen antibody libraries is to display proteins on the surface of yeast cells. Wittrup et al. (US Patents 6,699,658 and 6,696,251) developed a method for displaying libraries in yeast cells. In this yeast display system, one component includes the yeast lectin protein (Aga1), which is anchored to the yeast cell wall, and the other component includes the second subunit of the lectin protein Aga2, which can pass through disulfide bonds. Binds to the Aga1 protein and is displayed on the surface of yeast cells. Aga1 protein is expressed by integrating the Aga1 gene into the yeast chromosome. A single-chain variable fragment (scFv) library is fused to the Aga2 gene in a yeast display plasmid and, after transformation, is retained in yeast due to the presence of an additional nutritional marker. Both Aga1 and Aga2 proteins are expressed under the control of galactose-inducible promoters.
人抗体V基因库(VH和VK片段)是使用一组简并引物通过PCR方法获得(Sblattero，D.and Bradbury,A.Immunotechnology 3,271-2781998)。PCR模板来自可商购的RNA或cDNA，包括PBMC，脾脏，淋巴结，骨髓和扁桃体。将独立的VH和VKPCR文库合并后，通过重叠延伸PCR将其组装成scFv形式(Sheets，M.D.et al,Proc.Natl.Acad.Sci.USA 95,6157–61621998)。为了构建酵母scFv展示文库，通过同源重组将所得的scFv PCR产物克隆到酵母中的酵母展示质粒中。(Chao,G,et al,Nat Protoc.2006；1(2):755-68.Miller KD,et al.Current Protocols in Cytometry 4.7.1-4.7.30,2008)。The human antibody V gene library (V H and V K fragments) was obtained by PCR using a set of degenerate primers (Sblattero, D. and Bradbury, A. Immunotechnology 3, 271-278 1998). PCR templates were derived from commercially available RNA or cDNA, including PBMC, spleen, lymph nodes, bone marrow, and tonsils. Independent V H and V K PCR libraries were combined and assembled into scFv formats by overlap extension PCR (Sheets, MD et al, Proc. Natl. Acad. Sci. USA 95, 6157–61621998). To construct a yeast scFv display library, the resulting scFv PCR product was cloned into a yeast display plasmid in yeast via homologous recombination. (Chao, G, et al, Nat Protoc. 2006; 1(2):755-68. Miller KD, et al. Current Protocols in Cytometry 4.7.1-4.7.30, 2008).
可以利用哺乳动物细胞展示系统来筛选抗C5a抗体，其中抗体部分展示在细胞表面上并通过抗原导向的筛选方法分离出特异性靶向C5a的抗体(如U.S.patent No.7,732,195B2中所述)。可以建立展示大量人类IgG抗体基因的中国仓鼠卵巢(CHO)细胞文库，并将其用于发现表达高亲和力抗体基因的克隆。已开发出另一种展示系统，该系统通过可变剪接使同一蛋白同时在细胞表面展示和分泌，其中展示的蛋白表型保持与基因型相关，使得可同时在生物物理和基于细胞功能的分析中表征该分泌的可溶性抗体。该方法克服了先前哺乳动物细胞展示的许多局限性，能够直接筛选和成熟化全长的、糖基化的IgGs形式的抗体(PeterM.Bowers,et al,Methods 2014,65：44-56)。瞬时表达系统适用于在抗体基因恢复之前进行的单轮抗原选择，因此对于从较小文库中选择抗体最有用。稳定的外显体载体提供了一种有吸引力的选择。外显体载体可以高效转染并稳定地维持在低拷贝数，从而允许多轮淘选以及更复杂抗体库的解析。Anti-C5a antibodies can be screened using a mammalian cell display system, in which the antibody moiety is displayed on the cell surface and antibodies specifically targeting C5a are isolated by antigen-directed screening methods (as described in U.S. patent No. 7,732,195B2). A Chinese hamster ovary (CHO) cell library displaying a large number of human IgG antibody genes can be created and used to discover clones expressing high-affinity antibody genes. An alternative display system has been developed that allows the same protein to be simultaneously displayed and secreted on the cell surface through alternative splicing, in which the phenotype of the displayed protein remains correlated with the genotype, allowing for simultaneous biophysical and cell function-based analyses. Characterize the secreted soluble antibodies. This method overcomes many of the limitations of previous mammalian cell displays and enables direct screening and maturation of antibodies in the form of full-length, glycosylated IgGs (Peter M. Bowers, et al, Methods 2014, 65: 44-56). Transient expression systems are suitable for a single round of antigen selection prior to antibody gene recovery and are therefore most useful for selecting antibodies from smaller libraries. Stable exosome vectors offer an attractive option. Exosome vectors can be efficiently transfected and stably maintained at low copy numbers, allowing for multiple rounds of panning and elucidation of more complex antibody libraries.
IgG文库是基于分离自一群人类供体的种系序列V基因片段与重排的(D)J区域的连接构建而成。将从2000个人体血液样本中收集的RNA反转录为cDNA，使用VH和VK特异性引物扩增VH和VK片段，并通过凝胶提取纯化。将VH和VK片段分别亚克隆到包含IgG1或K恒定区的展示载体中，然后电穿孔或转导293T到细胞，从而制备IgG文库。为了制备scFv抗体展示文库，连接VH和VK以产生scFv，然后亚克隆到展示载体中，再将其电穿孔或转导293T细胞。众所周知，IgG文库是基于分离自一群供体的种系序列V基因片段与重排的(D)J区域构建而成，供体可以是小鼠，大鼠，兔或猴。The IgG library was constructed based on the ligation of germline sequence V gene fragments isolated from a pool of human donors and rearranged (D)J regions. RNA collected from 2000 human blood samples was reverse transcribed into cDNA, VH and VK fragments were amplified using VH and VK specific primers, and purified by gel extraction. The VH and VK fragments were subcloned into display vectors containing IgG1 or K constant regions respectively, and then electroporated or transduced 293T into cells to prepare IgG libraries. To prepare scFv antibody display libraries, V H and V K are ligated to generate scFv, which is then subcloned into a display vector and electroporated or transduced into 293T cells. As we all know, IgG libraries are constructed based on germline sequence V gene fragments and rearranged (D)J regions isolated from a group of donors, which can be mice, rats, rabbits or monkeys.
单克隆抗体也可以通过重组DNA方法进行制备，例如U.S.Patent No.4,816,567中所述。编码本申请中所述单克隆抗体的DNA可以通过常规方法(例如通过能特异性结合编码鼠源抗体轻链和重链基因的寡聚核苷酸探针)轻易的分离和测序。如上所述的杂交瘤细胞或本申请的C5a特异性噬菌体克隆可以作为这种DNA的来源。分离后，可将DNA置于表达载体中，然后该载体转染入宿主细胞，例如猿猴COS细胞、中华仓鼠卵巢癌(CHO)细胞或不产生免疫球蛋白的骨髓瘤细胞中，获得在重组宿主细胞中合成的单克隆抗体。所述DNA也可以被修饰，例如用编码序列取代人重链和轻链恒定区和/或用框架区替换同源非人序列(U.S.Patent No.4,816,567；Morrison et al.,supra)，或通过共价键连接免疫球蛋白的编码序列的全部或部分非免疫球蛋白多肽的编码序列。这种非免疫球蛋白多肽可以取代本申请中抗体的恒定区，或可以取代本申请中抗体可变结构域中的一个抗原结合位点，形成嵌合的二价抗体。Monoclonal antibodies can also be prepared by recombinant DNA methods, such as those described in U.S. Patent No. 4,816,567. DNA encoding the monoclonal antibodies described herein can be readily isolated and sequenced by conventional methods (eg, by oligonucleotide probes that specifically bind to genes encoding the light and heavy chains of murine antibodies). Hybridoma cells as described above or the C5a-specific phage clones of the present application can be used as a source of such DNA. After isolation, the DNA can be placed into an expression vector, which can then be transfected into host cells, such as simian COS cells, Chinese hamster ovary cancer (CHO) cells, or myeloma cells that do not produce immunoglobulins, to obtain the expression in the recombinant host. Monoclonal antibodies synthesized in cells. The DNA may also be modified, for example by substituting coding sequences for human heavy and light chain constant regions and/or substituting framework regions for homologous non-human sequences (U.S. Patent No. 4,816,567; Morrison et al., supra), or by All or part of the coding sequence for a non-immunoglobulin polypeptide is covalently linked to the coding sequence for an immunoglobulin. This non-immunoglobulin polypeptide can replace the constant region of the antibody in this application, or can replace an antigen-binding site in the variable domain of the antibody in this application, forming a chimeric bivalent antibody.
所述抗体可以是单价抗体。制备单价抗体的方法是本领域已知的。例如，一种涉及免疫球蛋白轻链和修饰重链的重组表达方法。通常在Fc区的任意位置截短重链，以阻止重链相互交联。或者，相关的半胱氨酸残基被其它氨基酸残基取代或被缺失以防止交联。The antibody may be a monovalent antibody. Methods of preparing monovalent antibodies are known in the art. For example, one involves a recombinant expression method involving an immunoglobulin light chain and a modified heavy chain. The heavy chain is usually truncated at any position in the Fc region to prevent the heavy chains from cross-linking with each other. Alternatively, the relevant cysteine residues are substituted with other amino acid residues or deleted to prevent cross-linking.
体外方法也适用于制备单价抗体。消化抗体产生抗体片段，特别是Fab片段，可以使用任何本领域已知的方法完成。In vitro methods are also suitable for preparing monovalent antibodies. Digestion of antibodies to produce antibody fragments, particularly Fab fragments, can be accomplished using any method known in the art.
具有所需结合特异性(抗体-抗原结合位点)的抗体可变结构域可以与免疫球蛋白恒定区融合。优选与免疫球蛋白重链恒定区进行融合，其包括至少部分铰链，CH2和CH3区。在一些实施例中，包含轻链结合必要位点的第一重链恒定区(CH1)至少出现在一种融合体中。编码免疫球蛋白重链融合体的DNA，如果需要，还可以包括编码免疫球蛋白轻链的DNA，被插入进独立的表达载体中，并共转染至合适的宿主生物中。Antibody variable domains with the desired binding specificity (antibody-antigen binding site) can be fused to immunoglobulin constant regions. Fusions to the immunoglobulin heavy chain constant region, including at least part of the hinge, CH2 and CH3 regions, are preferred. In some embodiments, the first heavy chain constant region (CH1), which contains the necessary sites for light chain binding, is present in at least one fusion. DNA encoding the immunoglobulin heavy chain fusion, and if desired, DNA encoding the immunoglobulin light chain, is inserted into a separate expression vector and co-transfected into a suitable host organism.
全人和人源化抗体Fully humanized and humanized antibodies
所述抗C5a抗体(如全长的抗C5a抗体)可以是全人抗体或人源化抗体。非人(如小鼠)抗体部分的人源化形式是嵌合的免疫球蛋白、免疫球蛋白链或其片段(例如Fv、Fab、Fab’、F(ab’)2、scFv或抗体的其他抗原结合子序列)，其通常包括最少的源于非人免疫球蛋白的序列。人源化抗体包括人免疫球蛋白、免疫球蛋白链或其片段(受体抗体)，其中受体CDR的残基被具有所需特异性、亲和力和性能的非人源(供体抗体)CDR残基取代，例如小鼠、大鼠或兔子的CDR。在一些实施例中，人免疫球蛋白Fv框架区残基被相应的非人源残基取代。人源化抗体还可以包含既不属于受体抗体也不在引入的CDR或框架区序列中的氨基酸残基。通常，人源化抗体包含至少一个，通常两个可变结构域，其中全部或基本上全部CDR区对应于非人免疫球蛋白的CDR区，全部或基本上全部框架区是人免疫球蛋白共有序列。The anti-C5a antibody (eg, full-length anti-C5a antibody) can be a fully human antibody or a humanized antibody. Humanized forms of non-human (e.g., mouse) antibody portions are chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (e.g., Fv, Fab, Fab', F(ab') 2 , scFv, or other fragments of antibodies Antigen binder sequences), which generally include minimal sequences derived from non-human immunoglobulins. Humanized antibodies include human immunoglobulins, immunoglobulin chains or fragments thereof (recipient antibodies) in which the residues of the acceptor CDRs are replaced by non-human (donor antibody) CDRs with the desired specificity, affinity and properties. Residue substitutions, such as mouse, rat or rabbit CDRs. In some embodiments, human immunoglobulin Fv framework residues are replaced with corresponding non-human residues. Humanized antibodies may also contain amino acid residues that neither belong to the recipient antibody nor are in the introduced CDR or framework sequence. Typically, a humanized antibody contains at least one, and usually two variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the framework regions are common to human immunoglobulins. sequence.
通常，人源化抗体含有一个或多个从非人源引入的氨基酸残基。那些非人源氨基酸残基通常被称为“移入”残基，通常来自“移入”可变结构域。根据一些实施例，人源化基本上可以按照Winter和其同事的如下方法进行(Jones et al.,Nature,321:522-525(1986)；Riechmann et al.,Nature,332:323-327(1988)；Verhoeyen et al.,Science,239:1534-1536(1988))，通过用啮齿动物CDRs或CDR序列取代人源抗体的相应序列。因此，这种“人源化”抗体部分(U.S.Patent No.4,816,567)，其基本上少于完整的人源抗体，其可变结构域已被来自非人源的相应序列所取代。在实际中，人源化抗体部分是典型的人源抗体部分，其中一些CDR残基和可能的一些框架区残基被来自啮齿类抗体中类似位点的残基所取代。Typically, humanized antibodies contain one or more amino acid residues introduced from a non-human source. Those non-human amino acid residues are often referred to as "implanted" residues, usually from the "imported" variable domain. According to some embodiments, humanization can be performed essentially as described by Winter and colleagues (Jones et al., Nature, 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 ( 1988); Verhoeyen et al., Science, 239:1534-1536 (1988)), by replacing the corresponding sequences of human antibodies with rodent CDRs or CDR sequences. Thus, this "humanized" antibody portion (U.S. Patent No. 4,816,567), which is substantially less than a fully human antibody, has its variable domains replaced by corresponding sequences from non-human sources. In practice, a humanized antibody portion is a typical human antibody portion in which some CDR residues and possibly some framework region residues are replaced by residues from similar positions in rodent antibodies.
全人抗体是人源化的一种替代方式。例如，目前可以制备在免疫后能够产生完整的全人抗体文库而不产生内源性免疫球蛋白的转基因动物(例如，小鼠)。例如，已有报道，嵌合和种系突变小鼠中抗体重链连接区(JH)基因的纯合子缺失，完全抑制了内源性抗体的产生。将人种系免疫球蛋白基因阵列转移到这种种系突变小鼠体内，可在抗原刺激下产生人源抗体，参见，例如akobovits et al.,PNAS USA,90:2551(1993)；Jakobovits et al.,Nature,362:255-258(1993)；Bruggemann et al.,Year in Immunol.,7:33(1993)；U.S.Patent Nos.5,545,806,5,569,825,5,591,669,5,545,807；和WO 97/17852。或者，可以通过将人类免疫球蛋白基因座引入转基因动物中(例如内源性免疫球蛋白基因已经被部分或全部沉默的小鼠)来制备全人抗体。抗原刺激后，可以发现全人抗体的产生在各个方面都与其在人类中的产生非常相似，包括基因重排、组装和抗体文库。这种方法在例如U.S.Patent Nos.5,545,807；5,545,806；5,569,825；5,625,126；5,633,425；and 5,661,016,and Marks et al.,Bio/Technology,10:779-783(1992)；Lonberg et al.,Nature,368:856-859(1994)；Morrison,Nature,368:812-813(1994)；Fishwild et al.,Nature Biotechnology,14:845-851(1996)；Neuberger,Nature Biotechnology,14:826(1996)；Lonberg and Huszar,Intern.Rev.Immunol.,13:65-93(1995)中进行了描述。Fully human antibodies are an alternative to humanization. For example, it is now possible to generate transgenic animals (eg, mice) that are capable of producing a complete library of fully human antibodies upon immunization without producing endogenous immunoglobulins. For example, it has been reported that homozygous deletion of the antibody heavy chain junction region (JH) gene in chimeric and germline mutant mice completely inhibits endogenous antibody production. Transferring human germline immunoglobulin gene arrays into such germline mutant mice can produce human antibodies in response to antigen stimulation, see, e.g., akobovits et al., PNAS USA, 90:2551 (1993); Jakobovits et al ., Nature, 362:255-258 (1993); Bruggemann et al., Year in Immunol., 7:33 (1993); U.S. Patent Nos. 5,545,806, 5,569,825, 5,591,669, 5,545,807; and WO 97/17852. Alternatively, fully human antibodies can be prepared by introducing human immunoglobulin loci into transgenic animals (eg, mice in which the endogenous immunoglobulin genes have been partially or completely silenced). After antigen stimulation, the production of fully human antibodies can be found to be very similar to their production in humans in all aspects, including gene rearrangement, assembly, and antibody libraries. This method is used in, for example, U.S. Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016, and Marks et al., Bio/Technology, 10:779-783 (1992); Lonberg et al., Nature, 368: 856-859(1994); Morrison, Nature, 368:812-813 (1994); Fishwild et al., Nature Biotechnology, 14: 845-851 (1996); Neuberger, Nature Biotechnology, 14: 826 (1996); Lonberg and Huszar, Intern. Rev. Immunol., 13:65-93 (1995).
全人抗体也以通过体外活化B细胞(见U.S.Patents 5,567,610 and 5,229,275)或通过使用本领域已知的各种技术来产生，包括噬菌体展示文库。Hoogenboom and Winter,J.Mol.Biol.,227:381(1991)；Marks et al.,J.Mol.Biol.,222:581(1991).Cole et al.和Boerner et al.等人的技术也可以用于制备全人单克隆抗体。见Cole et al.,Monoclonal Antibodies and Cancer Therapy,Alan R.Liss,p.77(1985)and Boerner et al.,J.Immunol.,147(1):86-95(1991)。Fully human antibodies can also be produced by activating B cells in vitro (see U.S. Patents 5,567,610 and 5,229,275) or by using various techniques known in the art, including phage display libraries. Hoogenboom and Winter, J. Mol. Biol., 227: 381 (1991); Marks et al., J. Mol. Biol., 222: 581 (1991). The technology of Cole et al. and Boerner et al. It can also be used to prepare fully human monoclonal antibodies. See Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985) and Boerner et al., J. Immunol., 147(1):86-95 (1991).
抗C5a抗体变体Anti-C5a antibody variants
在一些实施例中，本文提供的抗C5a抗体变体(例如，全长的抗C5a抗体)的氨基酸序列也在考虑中。例如，可能需要改善抗体的结合亲和力和/或其它生物学活性。抗体变体的氨基酸序列可以通过在编码抗体的核苷酸序列中引入适当的修饰或通过肽合成来制备。此类修饰包括例如，抗体氨基酸序列中残基的缺失和/或插入和/或取代。可以通过氨基酸残基缺失、插入和取代的任一组合来完成最终的构建，使其具有所需的特征。例如，抗原结合性。In some embodiments, the amino acid sequences of anti-C5a antibody variants (eg, full-length anti-C5a antibodies) provided herein are also contemplated. For example, it may be desirable to improve the binding affinity and/or other biological activity of the antibody. The amino acid sequences of antibody variants can be prepared by introducing appropriate modifications into the nucleotide sequence encoding the antibody or by peptide synthesis. Such modifications include, for example, deletions and/or insertions and/or substitutions of residues in the antibody amino acid sequence. The final construct can be accomplished by any combination of deletions, insertions, and substitutions of amino acid residues to give it the desired characteristics. For example, antigen binding.
在一些实施例中，提供具有一个或多个氨基酸取代的抗C5a抗体变体。取代突变的目标位点包括高变区(HVRs)和框架区(FRs)。可以在目标抗体中引入氨基酸取代，筛选所需活性的产物，例如，改善的生物活性，保持/改善抗原结合能力，降低的免疫原性，或改善的ADCC或CDC。In some embodiments, anti-C5a antibody variants with one or more amino acid substitutions are provided. Target sites for substitution mutations include hypervariable regions (HVRs) and framework regions (FRs). Amino acid substitutions can be introduced into the antibody of interest and the product screened for the desired activity, for example, improved biological activity, maintained/improved antigen binding capacity, reduced immunogenicity, or improved ADCC or CDC.
保守取代如下表4所示。Conservative substitutions are shown in Table 4 below.
表4保守取代



Table 4 Conservative substitutions



根据侧链性质将氨基酸分为不同类别：Amino acids are divided into different categories based on the nature of their side chains:
a、疏水氨基酸：去甲亮氨酸Norleucine、蛋氨酸Met、丙氨酸Ala、缬氨酸Val、亮氨酸Leu、异亮氨酸Ile；a. Hydrophobic amino acids: Norleucine, Met, Ala, Val, Leu, Ile;
b、中性亲水性氨基酸：半胱氨酸Cys、丝氨酸Ser、苏氨酸Thr、天冬酰胺Asn、谷氨酰胺Gln；b. Neutral hydrophilic amino acids: cysteine Cys, serine Ser, threonine Thr, asparagine Asn, glutamine Gln;
c、酸性氨基酸：天冬氨酸Asp、谷氨酸Glu；c. Acidic amino acids: aspartic acid Asp, glutamic acid Glu;
d、碱性氨基酸：组氨酸His、赖氨酸Lys、精氨酸Arg；d. Basic amino acids: histidine His, lysine Lys, arginine Arg;
e、含有影响链方向的氨基酸：甘氨酸Gly、脯氨酸Pro；e. Contains amino acids that affect chain direction: glycine Gly, proline Pro;
f、芳香族氨基酸：色氨酸Trp、酪氨酸Tyr、苯丙氨酸Phe。f. Aromatic amino acids: tryptophan Trp, tyrosine Tyr, phenylalanine Phe.
非保守氨基酸的取代包含将以上一种类别取代为另一种类别。Substitutions of non-conservative amino acids involve substitution of one category for another category.
一种示例性的取代变体是亲和力成熟的抗体，可采用例如以噬菌体展示为基础的亲和力成熟技术而方便地产生。简言之，将一个或多个CDR残基进行突变，变体抗体部分展示在噬菌体上，并筛选具有特定生物活性(例如，基于活性氧(ROS)释放试验的生物学活性或结合亲和力)的变体。可以在HVRs区进行改变(例如，取代)来获得改善的基于活性氧(ROS)释放试验的生物学活性或抗体亲和力。可以在HVR的“热点区”产生改变，即在体细胞成熟过程中发生高频突变的密码子编码的残基(参见，例如Chowdhury,Methods Mol.Biol.207:179-196(2008))，和/或在特异的决定性残基(SDRs)，检测所得变体VH和VL的结合亲和力。从二级文库中构建和重新选择亲和力成熟的方法已经在一些文献中进行描述，例如，Hoogenboom et al.in Methods in Molecular Biology 178:1-37(O'Brien et al.,ed.,Human Press,Totowa,NJ,(2001))。An exemplary substitution variant is an affinity matured antibody, which can be conveniently produced using, for example, phage display-based affinity maturation techniques. Briefly, one or more CDR residues are mutated, variant antibody moieties are displayed on phage, and those screened for specific biological activity (e.g., biological activity based on a reactive oxygen species (ROS) release assay or binding affinity) Variants. Alterations (eg, substitutions) can be made in regions of HVRs to obtain improved biological activity or antibody affinity based on reactive oxygen species (ROS) release assays. Alterations can occur in "hot spots" of HVR, i.e., codon-encoded residues that are highly mutated during somatic cell maturation (see, e.g., Chowdhury, Methods Mol. Biol. 207:179-196 (2008)), and/or detect the binding affinity of the resulting variants VH and VL at specific decisive residues (SDRs). Methods for constructing and reselecting affinity matured materials from secondary libraries have been described in the literature, e.g., Hoogenboom et al. in Methods in Molecular Biology 178:1-37 (O'Brien et al., ed., Human Press ,Totowa, NJ, (2001)).
在一些亲和力成熟的实施例中，通过多种方法中的任一种(例如易错PCR，链改组或寡核苷酸定向突变)，将多样性引入选择的用于亲和力成熟的可变基因中。然后创建二级文库。对该文库进行筛选，鉴定出具有所需亲和力的抗体变体。另一种引入多样性的方法包括HVR介导的方式，其中几个HVR残基(例如，一次4-6个残基)被随机化。涉及抗原结合的HVR残基被特异性地识别，例如，采用丙氨酸扫描诱变或建模。通常CDR-H3和CDR-L3区域尤其是重点靶标。In some affinity maturation embodiments, diversity is introduced into the variable genes selected for affinity maturation by any of a variety of methods, such as error-prone PCR, strand shuffling, or oligonucleotide-directed mutagenesis. . Secondary libraries are then created. The library is screened to identify antibody variants with the desired affinity. Another way to introduce diversity includes the HVR-mediated manner, in which several HVR residues (e.g., 4-6 residues at a time) are randomized. HVR residues involved in antigen binding are specifically recognized, for example, using alanine scanning mutagenesis or modeling. Usually the CDR-H3 and CDR-L3 regions are particularly focused targets.
在一些实施例中，取代、插入或缺失可能发生在一个或多个HVRs内，只要这种改变基本上不降低抗体结合抗原的能力。例如，可以在HVRs中产生基本上不降低结合亲和力的保守性改变(例如，本文中提供的保守性取代)。这些改变可能发生在HVR“热点区”或SDRs区域之外。在一些实施例中上文提供的变体VH和VL序列，每一个HVR或者是未发生改变，或者包含不超过1个、2个或3个氨基酸取代。In some embodiments, substitutions, insertions, or deletions may occur within one or more HVRs, so long as such changes do not substantially reduce the ability of the antibody to bind the antigen. For example, conservative changes (eg, conservative substitutions provided herein) can be made in HVRs that do not substantially reduce binding affinity. These changes may occur outside the HVR "hot zone" or SDRs area. In some embodiments of the variant VH and VL sequences provided above, each HVR is either unchanged or contains no more than 1, 2, or 3 amino acid substitutions.
一种有用的可以鉴定出抗体中能被靶向性突变的氨基酸残基或区域的方法称为“丙氨酸扫描突变”，如Cunningham and Wells(1989)Science,244:1081-1085中所述。在该方法中，一个或一组目标残基(例如，带电残基如精氨酸、天冬氨酸、组氨酸、赖氨酸和谷氨酸)被中性或带负电荷氨基酸(例如，丙氨酸或谷氨酸)取代，以此来确定抗体与抗原相互作用是否受到影响。可以在氨基酸的位置进一步引入取代，来证明该位置对初始取代具有功能敏感性。或者/另外，通过抗原-抗体复合物的晶体结构来鉴定抗体和抗原之间的接触位点。这些接触位点残基和邻近残基可作为取代候选物而被靶向或消除。筛选变体，确定它们是否具有所需要的性质。A useful method for identifying amino acid residues or regions of an antibody that can be targeted mutated is called "alanine scanning mutagenesis" as described in Cunningham and Wells (1989) Science, 244:1081-1085 . In this method, one or a group of target residues (e.g., charged residues such as arginine, aspartic acid, histidine, lysine, and glutamic acid) are replaced by neutral or negatively charged amino acids (e.g., , alanine or glutamic acid) substitution to determine whether the interaction between the antibody and the antigen is affected. Further substitutions can be introduced at amino acid positions to demonstrate functional sensitivity of the position to the initial substitution. Alternatively/in addition, the contact sites between the antibody and the antigen are identified through the crystal structure of the antigen-antibody complex. These contact site residues and adjacent residues can be targeted or eliminated as substitution candidates. Screen variants to determine if they have the desired properties.
氨基酸序列的插入，包括在氨基端和/或羧基末端的融合，长度范围从1个残基到包含100个或更多个残基的多肽，还包括在序列内插入1个或多个氨基酸残基。末端插入的例子包括N末端具有甲硫氨酰残基的抗体。抗体分子的其它插入变体，包括在抗体分子N-末端或C-末端融合一个酶(例如，ADEPT)或增加抗体血清半衰期的多肽。Insertions of amino acid sequences, including fusions at the amino and/or carboxyl terminus, ranging in length from 1 residue to polypeptides containing 100 or more residues, and including insertion of 1 or more amino acid residues within the sequence base. Examples of terminal insertions include antibodies with a methionyl residue at the N-terminus. Other insertional variants of antibody molecules include fusion of an enzyme (eg, ADEPT) or peptides that increase the serum half-life of the antibody to the N- or C-terminus of the antibody molecule.
Fc区变体Fc region variants
在一些实施例中，将一个或多个氨基酸修饰引入本文所述的抗体(例如，全长抗C5a抗体或抗C5a抗体融合蛋白)的Fc区，从而产生Fc区变体。在一些实施例中，Fc区变体具有增强的ADCC效能，通常与结合Fc的受体(FcRs)有关。在一些实施例中，Fc区变体具有降低的ADCC效能。有很多关于Fc序列的改变或突变影响其效能的例子，例如，WO 00/42072和Shields et al.J Biol.Chem.9(2):6591-6604(2001)描述了与FcRs的结合增强或减弱的抗体变体。这些出版物的内容通过引用并入本文。In some embodiments, one or more amino acid modifications are introduced into the Fc region of an antibody described herein (eg, a full-length anti-C5a antibody or an anti-C5a antibody fusion protein), thereby generating Fc region variants. In some embodiments, Fc region variants have enhanced ADCC potency, typically associated with binding to Fc receptors (FcRs). In some embodiments, Fc region variants have reduced ADCC efficacy. There are many examples of changes or mutations in the Fc sequence affecting its potency. For example, WO 00/42072 and Shields et al. J Biol. Chem. 9(2):6591-6604 (2001) describe enhanced binding to FcRs or Attenuated antibody variants. The contents of these publications are incorporated herein by reference.
抗体依赖的细胞介导的细胞毒作用(ADCC)是治疗性抗体针对肿瘤细胞的作用机制。ADCC是细胞介导的免疫防御，当靶细胞膜表面的抗原被特异性抗体(例如，抗C5a抗体)结合，免疫系统的效应细胞主动裂解靶细胞(例如，癌细胞)。通常ADCC效应涉及由抗体激活的NK细胞。NK细胞表达Fc受体CD16。该受体识别并结合与靶细胞表面相结合的抗体分子的Fc部分。NK细胞表面最常见的Fc受体为CD16或FcγRIII。Fc受体与抗体Fc区的结合导致NK细胞的活化，释放细胞裂解颗粒，随后靶细胞凋亡。ADCC对肿瘤细胞的杀伤作用可以通过转染高亲和力FcR的NK-92细胞的特异性实验来测定。其结果与不表达FcR的野生型NK-92进行比较。Antibody-dependent cell-mediated cytotoxicity (ADCC) is the mechanism of action of therapeutic antibodies against tumor cells. ADCC is a cell-mediated immune defense. When the antigen on the surface of the target cell membrane is bound by a specific antibody (for example, anti-C5a antibody), the effector cells of the immune system actively lyse the target cell (for example, cancer cells). Typically ADCC effects involve NK cells activated by antibodies. NK cells express the Fc receptor CD16. This receptor recognizes and binds to the Fc portion of the antibody molecule bound to the surface of the target cell. The most common Fc receptors on the surface of NK cells are CD16 or FcγRIII. Binding of Fc receptors to the Fc region of antibodies results in activation of NK cells, release of cell lytic granules, and subsequent target cell apoptosis. The killing effect of ADCC on tumor cells can be measured through specific experiments of NK-92 cells transfected with high-affinity FcR. The results were compared with wild-type NK-92, which does not express FcR.
在一些实施例中，本申请还提供抗C5a抗体变体(例如全长抗C5a抗体变体)，其包含具有部分但不是全部的效应功能Fc区，使得其在体内具有延长的半衰期，然而特定的效应功能(例如CDC或ADCC)是非必需的或有害的，这种抗C5a抗体成为本申请理想的候选。通过在体外和/或体内进行细胞毒性检测来确认CDC和/或ADCC活性的减少/消除。例如，通过Fc受体(FcR)结合试验来确认抗体缺乏FcγR结合能力(因此可能缺乏ADCC活性)但依然保留FcRn的结合能力。介导ADCC的主要细胞中，NK细胞仅表达FcγRIII，而单核细胞表达FcγRI、FcγRII和FcγRIII。Ravetch and Kinet Annu.Rev.Immunol.9:457-492(1991)第464页的表3中总结了FcR在造血细胞上的表达。在体外评估目标分子的ADCC活性的非限制性实例在U.S.Pat.No.5,500,362中进行了描述(参见例如Hellstrom,I.et al.Proc.Nat'l Acad.Sci.USA 83:7059-7063(1986)and Hellstrom,I et al.,Proc.Nat'l Acad.Sci.USA 82:1499-1502(1985)；U.S.Pat.No.5,821,337(see Bruggemann,M.et al.,J.Exp.Med.166:1351-1361(1987))。或者，可以采用非放射性检测方法(参见，例如ACTITM流式细胞术非放射性细胞毒性检测(CellTechnology,Inc.Mountain View,Calif.)和CYTOTOX 96TM非放射性细胞毒性检测(Promega,Madison,Wis.))。此类检测实验采用的效应细胞包括外周血单核细胞(PBMC)和自然杀伤细胞(NK)。或者，另外地，目标分子的ADCC活性在体内进行检测，例如，在动物模型中，如Clynes et al.Proc.Nat'l Acad.Sci.USA 95:652-656(1998)中所述。同时还可以进行C1q结合试验来确认抗体不能与C1q结合，从而缺乏CDC活性。参见，例如WO 2006/029879和WO 2005/100402中C1q和C3c结合ELISA。为了评估补体激活情况，可进行CDC检测(参见，例如Gazzano-Santoro et al.,J.Immunol.Methods 202:163(1996)；Cragg,M.S.et al.,Blood 101:1045-1052(2003)；和Cragg,M.S.and M.J.Glennie,Blood 103:2738-2743(2004))。使用本领域已知的方法来测定FcRn结合和体内清除/半衰期(参见，例如，Petkova,S.B.et al.,Int'l.Immunol.18(12):1759-1769(2006))。In some embodiments, the application also provides anti-C5a antibody variants (e.g., full-length anti-C5a antibody variants) that comprise an Fc region with some, but not all, of the effector functions such that they have an extended half-life in vivo, yet specifically The effector function (such as CDC or ADCC) is non-essential or deleterious, making this anti-C5a antibody an ideal candidate for this application. Reduction/elimination of CDC and/or ADCC activity is confirmed by performing cytotoxicity assays in vitro and/or in vivo. For example, Fc receptor (FcR) binding assays are used to confirm that the antibody lacks FcγR binding ability (and therefore may lack ADCC activity) but still retains FcRn binding ability. Among the main cells that mediate ADCC, NK cells only express FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. The expression of FcR on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet Annu. Rev. Immunol. 9:457-492 (1991). Non-limiting examples of in vitro assessment of ADCC activity of target molecules are described in US Pat. No. 5,500,362 (see, e.g., Hellstrom, I. et al. Proc. Nat'l Acad. Sci. USA 83:7059-7063 (1986 ) and Hellstrom, I et al., Proc. Nat'l Acad. Sci. USA 82:1499-1502 (1985); US Pat. No. 5,821,337 (see Bruggemann, M. et al., J. Exp. Med. 166 :1351-1361 (1987)). Alternatively, nonradioactive detection methods can be used (see, e.g., ACTI ™ Flow Cytometry Nonradioactive Cytotoxicity Assay (Cell Technology, Inc. Mountain View, Calif.) and CYTOTOX 96 ™ Nonradioactive Cell Toxicity assays (Promega, Madison, Wis.). Effector cells used in these assays include peripheral blood mononuclear cells (PBMC) and natural killer cells (NK). Alternatively, the ADCC activity of the target molecule is performed in vivo Detection, for example, in animal models, as described in Clynes et al. Proc. Nat'l Acad. Sci. USA 95:652-656 (1998). A C1q binding assay can also be performed to confirm that the antibody cannot bind to C1q , thereby lacking CDC activity. See, for example, C1q and C3c binding ELISA in WO 2006/029879 and WO 2005/100402. To assess complement activation, a CDC assay can be performed (see, for example, Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996); Cragg, MS et al., Blood 101:1045-1052 (2003); and Cragg, MS and MJ Glennie, Blood 103:2738-2743 (2004)). Determined using methods known in the art FcRn binding and clearance/half-life in vivo (see, eg, Petkova, SB et al., Int'l. Immunol. 18(12):1759-1769 (2006)).
具有降低的效应功能的抗体，包括在Fc区残基238、265、269、270、297、327和329位进行一个或多个残基的取代(U.S.Pat.No.6,737,056)。这些Fc变体包括在265、269、270、297和327位进行两个或多个残基的取代的Fc变体，包括被称为“DANA”的Fc变体，其在265和297位残基取代为丙氨酸(U.S.Pat.No.7,332,581)。Antibodies with reduced effector function include substitution of one or more residues at residues 238, 265, 269, 270, 297, 327, and 329 of the Fc region (U.S. Pat. No. 6,737,056). These Fc variants include Fc variants with substitutions of two or more residues at positions 265, 269, 270, 297 and 327, including the Fc variant known as "DANA" which has substitutions at residues 265 and 297. The base is substituted with alanine (U.S. Pat. No. 7,332,581).
这类与FcRs结合能力提高或降低的抗体变体已有描述(参见例如U.S.Pat.No.6,737,056；WO 2004/056312，和Shields et al.,J.Biol.Chem.9(2):6591-6604(2001))。Such antibody variants with increased or decreased binding ability to FcRs have been described (see, e.g., U.S. Pat. No. 6,737,056; WO 2004/056312, and Shields et al., J. Biol. Chem. 9(2):6591- 6604(2001)).
在一些实施例中，提供一种抗C5a抗体(例如全长的抗C5a抗体)变体，其包含具有一个或多个能够增强ADCC效应的氨基酸取代的Fc区变体。在一些实施例中，Fc区变体包含一个或多个能够增强ADCC效应的氨基取代，这些取代的位置在Fc区的298、333和/或334位(EU残基编号)。在一些实施例中，所述抗C5a抗体(例如，全长的抗C5a抗体)变体包括在Fc区的S298A，E333A和K334A位氨基酸取代。In some embodiments, an anti-C5a antibody (eg, a full-length anti-C5a antibody) variant is provided that includes an Fc region variant with one or more amino acid substitutions capable of enhancing ADCC effects. In some embodiments, the Fc region variant contains one or more amino substitutions capable of enhancing the ADCC effect at positions 298, 333 and/or 334 (EU residue numbering) of the Fc region. In some embodiments, the anti-C5a antibody (eg, full-length anti-C5a antibody) variant includes amino acid substitutions at positions S298A, E333A, and K334A in the Fc region.
在一些实施例中，Fc区的改变导致C1q结合和/或补体依赖性细胞毒作用(CDC)的改变(即增强或减弱)，参见U.S.Pat.No.6,194,551,WO 99/51642，和Idusogie et al.,J.Immunol.164:4178-4184(2000)中所述。In some embodiments, changes in the Fc region result in changes (i.e., enhancement or attenuation) of Clq binding and/or complement-dependent cytotoxicity (CDC), see U.S. Pat. No. 6,194,551, WO 99/51642, and Idusogie et al. al., J. Immunol. 164:4178-4184 (2000).
在一些实施例中，提供一种抗C5a抗体(例如全长的抗C5a抗体)变体，其包含具有一个或多个氨基酸取代的Fc区变体，能够延长半衰期或增强与Fc受体(FcRn)的结合。具有延长半衰期和改善FcRn结合的抗体在US 2005/0014934A1(Hinton等)中有所描述。这些抗体Fc区包含一个或多个氨基酸取代，增强了Fc区与FcRn的结合。这些Fc变体在Fc区包含238、256、265、272、286、303、305、307、311、312、317、340、356、360、362、376、378、380、382、413、424或434位的残基中的一个或多个取代，例如Fc区434位残基的取代(U.S.Pat.No.7,371,826)。In some embodiments, an anti-C5a antibody (e.g., a full-length anti-C5a antibody) variant is provided that includes an Fc region variant with one or more amino acid substitutions capable of extending half-life or enhancing interaction with an Fc receptor (FcRn ) combination. Antibodies with extended half-life and improved FcRn binding are described in US 2005/0014934A1 (Hinton et al.). The Fc region of these antibodies contains one or more amino acid substitutions that enhance the binding of the Fc region to FcRn. These Fc variants contain 238, 256, 265, 272, 286, 303, 305, 307, 311, 312, 317, 340, 356, 360, 362, 376, 378, 380, 382, 413, 424 or One or more substitutions in residue 434, for example, substitution of residue 434 in the Fc region (U.S. Pat. No. 7,371,826).
同时参见Duncan&Winter,Nature 322:738-40(1988)；U.S.Pat.No.5,648,260；U.S.Pat.No.5,624,821和WO 94/29351中提供其它Fc区变体的例子。See also Duncan & Winter, Nature 322:738-40 (1988); U.S. Pat. No. 5,648,260; U.S. Pat. No. 5,624,821 and WO 94/29351 for other examples of Fc region variants.
本申请考虑了包括本文所述的任一种Fc变体或其组合的抗C5a抗体(例如全长抗C5a抗体)。This application contemplates anti-C5a antibodies (eg, full-length anti-C5a antibodies) that include any of the Fc variants described herein, or combinations thereof.
糖基化变体Glycosylation variants
在一些实施例中，对本文所提供的抗C5a抗体(例如全长抗C5a抗体)进行改变，以增加或降低抗NGF抗体糖基化的程度。通过改变抗NGF抗体或其多肽部分的氨基酸序列以此来增加或去除一个或多个糖基化位点，可以方便地实现添加或删除抗C5a抗体上的糖基化位点。In some embodiments, anti-C5a antibodies provided herein (eg, full-length anti-C5a antibodies) are altered to increase or decrease the extent of glycosylation of the anti-NGF antibody. Adding or deleting glycosylation sites on the anti-C5a antibody can be conveniently achieved by changing the amino acid sequence of the anti-NGF antibody or its polypeptide portion to add or remove one or more glycosylation sites.
其中抗C5a抗体包含Fc区，可以改变与其连接的糖。由哺乳动物细胞产生的天然抗体通常包含分支的双触角寡糖，该寡糖通常通过N-连接与Fc区CH2结构域Asn297连接，参见例如Wright et al.，TIBTECH 15:26-32(1997)。所述寡糖可包含多种糖类，例如甘露糖、N-乙酰氨基葡萄糖苷(GlcNAc)、半乳糖和唾液酸，以及与双触角寡糖结构“茎”部的GlcNAc相连接的海藻糖。在一些实施例中，可对本申请的抗C5a抗体进行寡糖修饰，从而产生具有某些改进特性的抗C5a抗体变体。The anti-C5a antibody contains an Fc region that can change the sugar attached to it. Natural antibodies produced by mammalian cells typically contain branched biantennary oligosaccharides that are often linked to the Fc region CH2 domain Asn297 via an N-link, see e.g. Wright et al., TIBTECH 15:26-32 (1997) . The oligosaccharides can include a variety of sugars, such as mannose, N-acetylglucosamine (GlcNAc), galactose, and sialic acid, as well as trehalose linked to GlcNAc in the "stem" portion of the biantennary oligosaccharide structure. In some embodiments, the anti-C5a antibodies of the present application can be subjected to oligosaccharide modifications, thereby producing anti-C5a antibody variants with certain improved properties.
与Fc区的CH2结构域连接的N-聚糖是异质的。CHO细胞中产生的抗体或Fc融合蛋白通过岩藻糖基转移酶活性被岩藻糖基化，参见Shoji-Hosaka et al.,J.Biochem.2006,140:777-83。通常，可以在人血清中检测出一小部分天然存在的非岩藻糖基化IgGs。Fc区的N-糖基化对于其与FcγR结合很重要；而非岩藻糖基化的N-聚糖增强了Fc与FcγRIIIa的结合能力。与FcRIIIa结合能力增强使得ADCC效应增强，这在需要细胞毒性的某些抗体治疗应用中是有利的。The N-glycans linked to the CH2 domain of the Fc region are heterogeneous. Antibodies or Fc fusion proteins produced in CHO cells are fucosylated by fucosyltransferase activity, see Shoji-Hosaka et al., J. Biochem. 2006, 140:777-83. Typically, a small proportion of naturally occurring afucosylated IgGs can be detected in human serum. N-glycosylation of the Fc region is important for its binding to FcγR; afucosylated N-glycan enhances the binding ability of Fc to FcγRIIIa. The enhanced binding ability to FcRIIIa results in enhanced ADCC effect, which is advantageous in certain antibody therapeutic applications that require cytotoxicity.
在一些实施例中，当不需要Fc介导的细胞毒作用时，增强的效应功能可能是有害的。在一些实施例中，Fc片段或CH2结构域是非糖基化的。在一些实施例中，通过对CH2结构域中的N-糖基化位点进行突变以阻止其糖基化。In some embodiments, enhanced effector function may be detrimental when Fc-mediated cytotoxicity is not required. In some embodiments, the Fc fragment or CH2 domain is non-glycosylated. In some embodiments, glycosylation is prevented by mutating the N-glycosylation site in the CH2 domain.
在一些实施例中，提供抗C5a抗体(例如全长的抗C5a抗体)变体，其包含Fc区，其中连接于Fc区的糖类结构具有减少的岩藻糖或缺乏岩藻糖，这可能会增强ADCC功能。具体地，本文提供抗C5a抗体，其相对于野生型CHO细胞产生的相同抗C5a抗体具有减少的岩藻糖。也就是说，它们的特征在于，与天然CHO细胞(例如，产生天然糖基化形式的CHO细胞，含有天然FUT8基因的CHO细胞)产生的抗体相比，具有更少量的岩藻糖。在一些实施例中，所述抗C5a抗体的N-连接聚糖具有少于50％、40％、30％、20％、10％或5％的岩藻糖。例如，该抗C5a抗体的岩藻糖含量可能是1％-80％、1％-65％、5％-65％或20％-40％。在一些实施例中，所述抗C5a抗体的N-连接聚糖不包含岩藻糖，即，其中抗C5a抗体完全不含岩藻糖，或没有岩藻糖或是去岩藻糖基化。岩藻糖的含量是通过计算连接到Asn297上的糖链内岩藻糖平均含量相对于通过MALDI-TOF质谱测量的所有连接在Asn297上的糖结构(如复合、杂交或甘露糖结构)的总量来确定的，如WO 2008/077546所述。Asn297是指位于Fc区297位的天冬酰胺残基(EU Fc区残基编号体系)。然而，由于抗体的微小序列变化，Asn297也可位于297位的上游或下游±3个氨基酸，即在294和300位之间。这些岩藻糖基化变体可能具有增强的ADCC功能。参见例如US Patent Publication Nos.US 2003/0157108(Presta,L.)，US 2004/0093621(Kyowa Hakko Kogyo Co.,Ltd)。与“去岩藻糖基化”或“岩藻糖缺乏”的抗体变体相关的出版物的实例包括，US 2003/0157108；WO 2000/61739；WO 2001/29246；US 2003/0115614；US 2002/0164328；US 2004/0093621；US 2004/0132140；US 2004/0110704；US 2004/0110282；US 2004/0109865；WO 2003/085119；WO 2003/084570；WO 2005/035586；WO 2005/035778；WO 2005/053742；WO 2002/031140；Okazaki et al.J.Mol.Biol.336:1239-1249(2004)；Yamane-Ohnuki et al.Biotech.Bioeng.87:614(2004)。能够产生去岩藻糖基化抗体的细胞系包括缺乏蛋白岩藻糖基化功能的Lec13 CHO细胞(Ripka et al.Arch.Biochem.Biophys.249:533-545(1986)；US PatAppl No US 2003/0157108 A1,Presta,L；和WO 2004/056312 A1,Adams et al.,尤其是实施例11)，和基因敲除细胞系，例如α-1,6-岩藻糖基转移酶基因，FUT8基因敲除的CHO细胞(参见Yamane-Ohnuki et al.Biotech.Bioeng.87:614(2004)；Kanda,Y.et al.,Biotechnol.Bioeng.,94(4):680-688(2006)；和WO2003/085107)。In some embodiments, anti-C5a antibody (eg, full-length anti-C5a antibody) variants are provided that comprise an Fc region in which the carbohydrate structure linked to the Fc region has reduced fucose or lacks fucose, which may Will enhance ADCC function. Specifically, provided herein are anti-C5a antibodies that have reduced fucose relative to the same anti-C5a antibody produced by wild-type CHO cells. That is, they are characterized by having smaller amounts of fucose than antibodies produced by native CHO cells (e.g., CHO cells that produce the native glycosylated form, CHO cells that contain the native FUT8 gene). In some embodiments, the N-linked glycan of the anti-C5a antibody has less than 50%, 40%, 30%, 20%, 10%, or 5% fucose. For example, the fucose content of the anti-C5a antibody may be 1%-80%, 1%-65%, 5%-65%, or 20%-40%. In some embodiments, the N-linked glycans of the anti-C5a antibody do not comprise fucose, i.e., wherein the anti-C5a antibody contains no fucose at all, or is free of fucose or afucosylated. The fucose content is calculated by calculating the average fucose content within the sugar chain attached to Asn297 relative to the total of all sugar structures attached to Asn297 (such as complex, hybrid or mannose structures) measured by MALDI-TOF mass spectrometry. determined by quantity, as described in WO 2008/077546. Asn297 refers to the asparagine residue located at position 297 in the Fc region (EU Fc region residue numbering system). However, due to minor sequence changes in the antibody, Asn297 can also be located ±3 amino acids upstream or downstream of position 297, i.e., between positions 294 and 300. These fucosylation variants may have enhanced ADCC functions. See, for example, US Patent Publication Nos. US 2003/0157108 (Presta, L.), US 2004/0093621 (Kyowa Hakko Kogyo Co., Ltd). Examples of publications related to "afucosylated" or "fucose-deficient" antibody variants include, US 2003/0157108; WO 2000/61739; WO 2001/29246; US 2003/0115614; US 2002 /0164328; US 2004/0093621; US 2004/0132140; US 2004/0110704; US 2004/0110282; US 2004/0109865; WO 2003/085119; WO 2003/084570; WO 2005/035 586;WO 2005/035778;WO 2005 /053742; WO 2002/031140; Okazaki et al. J. Mol. Biol. 336: 1239-1249 (2004); Yamane-Ohnuki et al. Biotech. Bioeng. 87: 614 (2004). Cell lines capable of producing afucosylated antibodies include Lec13 CHO cells lacking protein fucosylation function (Ripka et al. Arch. Biochem. Biophys. 249:533-545 (1986); US PatAppl No US 2003 /0157108 A1, Presta, L; and WO 2004/056312 A1, Adams et al., especially Example 11), and gene knockout cell lines, such as α-1,6-fucosyltransferase gene, FUT8 Gene knockout CHO cells (see Yamane-Ohnuki et al. Biotech. Bioeng. 87:614 (2004); Kanda, Y. et al., Biotechnol. Bioeng., 94 (4): 680-688 (2006); and WO2003/085107).
抗C5a抗体(例如全长抗C5a抗体)变体进一步提供二等分寡糖，例如，其中连接于抗C5a抗体Fc区的双触角寡糖被GlcNAc等分。这种抗C5a抗体(例如全长的抗C5a抗体)变体可能具有减少的岩藻糖基化和/或增强的ADCC功能。这类抗体变体的实例在WO 2003/011878(Jean-Mairet et al.)；U.S.Pat.No.6,602,684(Umana et al.)；US 2005/0123546(Umana et al.)，和Ferrara et al.,Biotechnology and Bioengineering,93(5):851-861(2006)中有所描述。还提供抗C5a抗体(例如全长的抗C5a抗体)变体，其在与Fc区连接的寡糖中具有至少一个半乳糖残基。这类抗C5a抗体变体可能具有增强的CDC功能。这类变体在例如WO 1997/30087(Patel et al.)；WO 1998/58964(Raju,S.)；和WO 1999/22764(Raju,S.)中有所描述。Anti-C5a antibody (eg, full-length anti-C5a antibody) variants further provide bisecting oligosaccharides, eg, wherein the biantennary oligosaccharide linked to the Fc region of the anti-C5a antibody is bisected by GlcNAc. Such anti-C5a antibody (eg, full-length anti-C5a antibody) variants may have reduced fucosylation and/or enhanced ADCC function. Examples of such antibody variants are in WO 2003/011878 (Jean-Mairet et al.); U.S. Pat. No. 6,602,684 (Umana et al.); US 2005/0123546 (Umana et al.), and Ferrara et al. Described in ,Biotechnology and Bioengineering, 93(5):851-861(2006). Also provided are anti-C5a antibody (eg, full-length anti-C5a antibody) variants that have at least one galactose residue in the oligosaccharide linked to the Fc region. Such anti-C5a antibody variants may have enhanced CDC function. Such variants are described, for example, in WO 1997/30087 (Patel et al.); WO 1998/58964 (Raju, S.); and WO 1999/22764 (Raju, S.).
在一些实施例中，所述抗C5a抗体(例如全长抗C5a抗体)变体包含能与FcγRIII结合的Fc区。在一些实施例中，包含Fc区的所述抗C5a抗体(例如全长抗C5a抗体)变体在人效应细胞(例如T细胞)存在下具有ADCC活性，或者与具有人野生型IgG1 Fc区的其他相同抗C5a抗体(例如全长抗C5a抗体)相比，在人效应细胞存在下，具有增强的ADCC活性。In some embodiments, the anti-C5a antibody (eg, full-length anti-C5a antibody) variant comprises an Fc region capable of binding FcγRIII. In some embodiments, the anti-C5a antibody (e.g., full-length anti-C5a antibody) variant comprising an Fc region has ADCC activity in the presence of human effector cells (e.g., T cells), or is identical to an anti-C5a antibody having a human wild-type IgG1 Fc region. Compared with other identical anti-C5a antibodies (eg, full-length anti-C5a antibodies), they have enhanced ADCC activity in the presence of human effector cells.
半胱氨酸工程变体Cysteine engineered variants
在一些实施例中，需要制备半胱氨酸工程化的抗C5a抗体(例如全长抗C5a抗体)，在该抗体中一个或多个氨基酸残基被半胱氨酸残基取代。在一些实施例中，取代残基出现在抗C5a抗体的可及位点。通过用半胱氨酸取代那些残基，具有活性的巯基基团位于抗C5a抗体的可及位点，可以用于将该抗C5a抗体与其它部分偶联，例如药物部分或接头-药物部分，来制备如本文中进一步描述的抗C5a免疫偶联物。半胱氨酸工程化的抗C5a抗体(例如，全长抗C5a抗体)可以按照例如U.S.Pat.No.7,521,541所述进行制备。In some embodiments, it is desirable to prepare a cysteine-engineered anti-C5a antibody (eg, a full-length anti-C5a antibody) in which one or more amino acid residues are replaced with cysteine residues. In some embodiments, the substitution residue occurs at an accessible site of the anti-C5a antibody. By replacing those residues with cysteine, reactive thiol groups are located in accessible sites of the anti-C5a antibody and can be used to couple the anti-C5a antibody to other moieties, such as a drug moiety or a linker-drug moiety, to prepare anti-C5a immunoconjugates as further described herein. Cysteine-engineered anti-C5a antibodies (eg, full-length anti-C5a antibodies) can be prepared, for example, as described in U.S. Pat. No. 7,521,541.
衍生物derivative
在一些实施例中，本文所提供的抗C5a抗体(例如全长抗C5a抗体)可进一步修饰以包含本领域已知并且容易获得的其它非蛋白部分。适用于衍生化抗C5a抗体的部分包括，但不限于，水溶性聚合物。水溶性聚合物的非限制性实例包括，但不限于，聚乙二醇(PEG)、乙二醇/丙二醇共聚物、羧甲基纤维素、右旋糖酐、聚乙烯醇、聚乙烯吡咯烷酮、聚-1,3-二氧戊烷、聚-1,3,6-三氧杂环已烷、乙烯/马来酸酐共聚物、聚氨基酸(均聚物或无规共聚物)、右旋糖酐或聚(n-乙烯基吡咯烷酮)聚乙二醇、丙二醇均聚物、环氧丙烷/环氧乙烷共聚物、聚氧乙基化多元醇(例如甘油)、聚乙烯醇及其混合物。聚乙二醇丙醛由于其在水中的稳定性，在制造中具有优势。聚合物可以具有任意分子量，可以是支链或非支链的。连接在抗C5a抗体上的聚合物数量可以变化，并且如果连接多于一个多聚物，它们可以是相同的或不同的分子。通常，用于衍生化的聚合物的数量和/或类型可基于以下考虑因素来确定，包括但不限于，需要改进抗C5a抗体的特性或功能，抗C5a抗体衍生物是否用于特定条件下的治疗等。In some embodiments, anti-C5a antibodies (eg, full-length anti-C5a antibodies) provided herein can be further modified to include other non-protein moieties known in the art and readily available. Suitable moieties for derivatizing anti-C5a antibodies include, but are not limited to, water-soluble polymers. Non-limiting examples of water-soluble polymers include, but are not limited to, polyethylene glycol (PEG), ethylene glycol/propylene glycol copolymer, carboxymethylcellulose, dextran, polyvinyl alcohol, polyvinylpyrrolidone, poly-1 , 3-dioxopentane, poly-1,3,6-trioxane, ethylene/maleic anhydride copolymer, polyamino acid (homopolymer or random copolymer), dextran or poly(n- vinylpyrrolidone) polyethylene glycol, propylene glycol homopolymer, propylene oxide/ethylene oxide copolymer, polyoxyethylated polyols (such as glycerol), polyvinyl alcohol, and mixtures thereof. Polyethylene glycol propionaldehyde offers advantages in manufacturing due to its stability in water. The polymers can be of any molecular weight and can be branched or unbranched. The number of polymers attached to the anti-C5a antibody can vary, and if more than one polymer is attached, they can be the same or different molecules. Generally, the amount and/or type of polymer used for derivatization can be determined based on considerations including, but not limited to, the need to improve the properties or functionality of the anti-C5a antibody, whether the anti-C5a antibody derivative is useful under specific conditions. Treatment etc.
药物组合物pharmaceutical composition
本文还提供包含任一种抗C5a抗体(例如全长抗C5a抗体)、编码抗体的核酸、包含编码抗体的核酸的载体或者包含本文所述的核酸或载体的宿主细胞的组合物(例如药物组合物，在这里也称为制剂)。在一些实施例中，提供一种药物组合物，包含本文所述的任一种抗C5a抗体和药学上可接受的载体。Also provided herein are compositions (e.g., pharmaceutical combinations) comprising any of the anti-C5a antibodies (eg, full-length anti-C5a antibodies), nucleic acids encoding the antibodies, vectors comprising nucleic acids encoding the antibodies, or host cells comprising the nucleic acids or vectors described herein. substances, also referred to here as preparations). In some embodiments, a pharmaceutical composition is provided comprising any of the anti-C5a antibodies described herein and a pharmaceutically acceptable carrier.
可通过混合具有所需纯度的抗C5a抗体与任选的药学上可接受的载体、赋形剂或稳定剂(Remington's Pharmaceutical Sciences 16th edition,Osol,A.Ed.(1980))获得合适的抗C5a抗体制剂，制备成冻干制剂或液体制剂形式。可接受的载体、赋形剂或稳定剂在所用剂量和浓度下对接受者无毒，包括缓冲剂如：磷酸盐、柠檬酸和其它有机酸；抗氧化剂，包括抗坏血酸和蛋氨酸；防腐剂(例如十八烷基二甲基苄基氯化铵；六甲基氯化铵；苯扎氯铵；苄索氯铵；苯酚；丁醇或苄醇；对羟基苯甲酸烷基酯，如对羟基苯甲酸甲酯或对羟基苯甲酸丙酯；邻苯二酚；间苯二酚；环己醇；3-戊醇和间甲酚)；低分子量(少于10个残基)多肽；蛋白质，例如血清白蛋白、明胶或免疫球蛋白；亲水性聚合物，如聚乙烯吡咯烷酮；氨基酸，例如甘氨酸、谷氨酰胺、天冬酰胺、组氨酸、精氨酸或赖氨酸；单糖、二糖和其它碳水化合物，包括葡萄糖、甘露糖或糊精；螯合剂如EDTA；糖类，如蔗糖、甘露醇、海藻糖或山梨糖醇；成盐反离子如钠；金属复合物(如锌-蛋白复合物)；和/或非离子表面活性剂如TWEENTM，PLURONICSTM或聚乙二醇(PEG)；示例性制剂如WO98/56418中所述，并通过引用明确并入本文。适合皮下给药的冻干制剂在WO97/04801中有所描述。这类冻干制剂可通过合适的稀释剂重构成高蛋白浓度的制剂，并且重构的制剂可以通过皮下给药的方式给予本文中待治疗个体。阳离子脂质体或脂质体可以用于将本申请中的抗C5a抗体递送至细胞。A suitable anti-C5a antibody can be obtained by mixing an anti-C5a antibody of the desired purity with an optional pharmaceutically acceptable carrier, excipient or stabilizer (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)) Antibody preparations are prepared in the form of lyophilized preparations or liquid preparations. Acceptable carriers, excipients or stabilizers are non-toxic to the recipient at the doses and concentrations used and include buffers such as phosphates, citric acid and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (e.g. Octadecyldimethylbenzyl ammonium chloride; hexamethylammonium chloride; benzalkonium chloride; benzethonium chloride; phenol; butanol or benzyl alcohol; alkyl parabens, such as p-hydroxybenzoate Methyl formate or propyl parahydroxybenzoate; catechol; resorcinol; cyclohexanol; 3-pentanol and m-cresol); low molecular weight (less than 10 residues) peptides; proteins, e.g. serum Albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates, including glucose, mannose or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counterions such as sodium; metal complexes such as zinc-protein complex); and/or non-ionic surfactants such as TWEEN ™ , PLURONICS ™ or polyethylene glycol (PEG); exemplary formulations are as described in WO98/56418, expressly incorporated herein by reference. Lyophilized formulations suitable for subcutaneous administration are described in WO97/04801. Such lyophilized formulations can be reconstituted with appropriate diluents into high protein concentration formulations, and the reconstituted formulations can be administered subcutaneously to the individuals to be treated herein. Cationic liposomes or liposomes can be used to deliver the anti-C5a antibodies of the present application to cells.
本文所述的制剂除包含抗C5a抗体(例如全长抗C5a抗体)之外，还可以包含一种或多种治疗特定病症所必要的其它活性物质，优选具有活性互补且彼此无不良反应的物质。例如，除了抗C5a抗体之外，可能需要进一步包含抗肿瘤剂、生长抑制剂、细胞毒剂或化学治疗试剂。这些分子以对预期目的有效的量组合存在。其它物质的有效量取决于制剂中的抗C5a抗体的含量，疾病或病症或治疗的类型，以及如上所述的其它因素。这些药物通常以与本文描述的相同剂量和给药途径使用，或者以目前应用剂量的1％至99％使用。In addition to anti-C5a antibodies (eg, full-length anti-C5a antibodies), the formulations described herein may also contain one or more other active substances necessary for the treatment of specific conditions, preferably substances with complementary activities and no adverse reactions to each other. . For example, in addition to the anti-C5a antibody, it may be desirable to further include antineoplastic agents, growth inhibitory agents, cytotoxic agents, or chemotherapeutic agents. These molecules are present in combination and in amounts effective for the intended purpose. Effective amounts of other substances will depend on the amount of anti-C5a antibody in the formulation, the type of disease or condition or treatment, and other factors as discussed above. These drugs are typically used at the same dosages and routes of administration as described herein, or at 1% to 99% of currently used dosages.
所述抗C5a抗体(例如，全长抗C5a抗体)也可以包埋在例如通过凝聚技术和界面聚合制备的微胶囊中，例如分别在胶体药物递送系统(例如，脂质体、白蛋白微球、微乳液、纳米颗粒和纳米胶囊)中或粗乳液中的羟甲基纤维素或明胶-微胶囊和聚(甲基丙烯酸甲酯)微胶囊。可以制备缓释制剂。The anti-C5a antibodies (e.g., full-length anti-C5a antibodies) can also be embedded in microcapsules prepared, for example, by coacervation techniques and interfacial polymerization, such as in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, respectively). , microemulsions, nanoparticles and nanocapsules) or in macroemulsions - microcapsules and poly(methyl methacrylate) microcapsules. Sustained release formulations can be prepared.
可以制备抗C5a抗体(例如，全长抗C5a抗体)的缓释制剂。缓释制剂的适合的实例包括含有抗体(或其片段)的固体疏水聚合物半透性基质，这些基质是成型制品的形式，例如，薄膜或微胶囊。缓释基质的实例包括聚酯、水凝胶(例如，聚(2-羟乙基甲基丙烯酸酯)或聚(乙烯醇))、聚乳酸(U.S.Pat.No.3,773,919)，L-谷氨酸和L-谷氨酸乙酯共聚物，不可降解的乙烯-醋酸乙烯酯，可降解的乳酸-乙醇酸共聚物如LUPRON DEPOTTM(由乳酸-乙醇酸共聚物和醋酸亮丙瑞林组成的可注射微球)以及聚-D(-)-3-羟基丁酸。虽然诸如乙烯-醋酸乙烯酯和乳酸-乙醇酸之类的聚合物可以使分子的释放超过100天，某些水凝胶可以在更短的时间内释放蛋白质。当包封的抗体在体内长时间停留时，它们会因暴露于37℃的潮湿环境中发生变性或聚集，可能导致生物活性的丧失或免疫原性的改变。可以根据相应的机制，设计合理的策略来稳定抗C5a抗体。例如，如果发现聚集机制是通过硫代二硫化物交换形成分子间S-S键，则可以通过修饰巯基残基、在酸性溶液中冻干、控制含水量、使用适当的添加剂、以及开发特定的聚合物基质组合物来实现稳定化。Sustained release formulations of anti-C5a antibodies (eg, full-length anti-C5a antibodies) can be prepared. Suitable examples of sustained release formulations include solid hydrophobic polymeric semipermeable matrices containing the antibodies (or fragments thereof) in the form of shaped articles, for example, films or microcapsules. Examples of sustained-release matrices include polyester, hydrogel (eg, poly(2-hydroxyethyl methacrylate) or poly(vinyl alcohol)), polylactic acid (U.S. Pat. No. 3,773,919), L-glutamine Acid and L-glutamic acid ethyl ester copolymer, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymer such as LUPRON DEPOTTM (a biodegradable product composed of lactic acid-glycolic acid copolymer and leuprolide acetate) injection microspheres) and poly-D(-)-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic-glycolic acid can release molecules over 100 days, certain hydrogels can release proteins in much shorter time periods. When encapsulated antibodies stay in the body for a long time, they will denature or aggregate due to exposure to a humid environment at 37°C, which may lead to loss of biological activity or changes in immunogenicity. Reasonable strategies can be designed to stabilize anti-C5a antibodies based on the corresponding mechanisms. For example, if the aggregation mechanism is found to be through thiodisulfide exchange to form intermolecular S-S bonds, this can be achieved by modifying sulfhydryl residues, lyophilizing in acidic solutions, controlling water content, using appropriate additives, and developing specific polymers. matrix composition to achieve stabilization.
在一些实施例中，所述抗C5a抗体(例如全长抗C5a抗体)配制在含有柠檬酸盐、氯化钠、乙酸盐、琥珀酸盐、甘氨酸、聚山梨酯80(吐温80)或上述任何组合的缓冲液中。In some embodiments, the anti-C5a antibody (eg, full-length anti-C5a antibody) is formulated with citrate, sodium chloride, acetate, succinate, glycine, polysorbate 80 (Tween 80), or in any combination of the above buffers.
用于体内给药的制剂必须是无菌的。这可以通过例如应用无菌过滤膜过滤而容易地实现。Preparations for in vivo administration must be sterile. This can be easily accomplished, for example, by applying sterile filtration membrane filtration.
使用抗C5a抗体的治疗方法Treatment using anti-C5a antibodies
抗C5a抗体(例如，全长的抗C5a抗体)和/或本申请所述的组合物可以施用于个体(例如，哺乳动物，如人类)来治疗与C5a信号通路失调导致的疾病和/或病症(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)，这些疾病包括但不限于炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。因此，本申请在一些实施例中，提供一种治疗C5a信号通路失调导致的疾病和/或病症(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的方法，包括向个体施用有效量的包含抗C5a抗体(例如，全长的抗C5a抗体)的组合物(例如，药物组合物)，例如本文所述的任一种抗C5a抗体(例如，全长的抗C5a抗体)，在一些实施例中，所述个体是人类。Anti-C5a antibodies (e.g., full-length anti-C5a antibodies) and/or compositions described herein may be administered to an individual (e.g., a mammal, such as a human) to treat diseases and/or conditions resulting from dysregulation of the C5a signaling pathway (e.g., autoimmune and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune dysregulation diseases), including but not limited to inflammatory response syndrome (SIRS), sepsis , severe sepsis, septic shock, ischemia/reperfusion-related injury, acute lung injury, pneumonia, acute and chronic graft rejection in transplant patients, graft-versus-host reaction, glomerular disease, glomerulonephritis, renal failure of entities, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth and solid organ cancers. Therefore, the present application, in some embodiments, provides a method for treating diseases and/or conditions caused by dysregulation of the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplantation for immune dysregulation diseases), comprising administering to an individual an effective amount of a composition (e.g., a pharmaceutical composition) comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), such as any of the methods described herein. An anti-C5a antibody (eg, a full-length anti-C5a antibody), in some embodiments, the subject is a human.
例如，在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含特异性结合人C5a上表位的C5a抗体(例如，全长抗C5a抗体)的药物组合物，其中所述表位包含人C5a的位氨基酸残基。在一些实施例中，所述抗C5a抗体是全长抗体。在一些实施例中，所述全长抗C5a抗体是IgG1或IgG4抗体。在一些实施例中，所述疾病或病症选自例如，炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症组成的组中。在一些实施例中，所述个体是人类。For example, in some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplantation). (immune dysregulation disease), comprising administering to the individual an effective amount of a pharmaceutical composition comprising a C5a antibody (e.g., a full-length anti-C5a antibody) that specifically binds an epitope on human C5a, wherein the epitope Contains the amino acid residues of human C5a. In some embodiments, the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody. In some embodiments, the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, In the group consisting of Crohn's disease, tumor growth, and solid organ cancers. In some embodiments, the individual is a human.
例如，在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体(例如，全长抗C5a抗体)的药物组合物，重链可变结构域(VH)，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:2，和HC-CDR3，其包含氨基酸序列SEQ ID NO:6，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及轻链可变结构域(VL)，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。在一些实施例中，所述抗C5a抗体是全长抗体。在一些实施例中，所述全长抗C5a抗体是IgG1或IgG4抗体。在一些实施例中，所述疾病或病症选自例如，炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。在一些实施例中，所述个体是人类。For example, in some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplantation). A method for an individual suffering from an immune dysregulation disease), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 2, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 6, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 12. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 20, or a variant of the V L , which contains up to about 5 LC-CDRs. Amino acid substitutions. In some embodiments, the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody. In some embodiments, the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers. In some embodiments, the individual is a human.
在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体的组合物，其中所述抗体包含：VH，所述VH包含SEQ ID NO:25所示的氨基酸序列或其变体，所述变体与SEQ ID NO:25所示的氨基酸序列具有至少约80％序列同一性；以及VL，所述VL包含SEQ ID NO:35所示的氨基酸序列或其变体，所述变体与SEQ ID NO:35所示的氨基酸序列具有至少约80％序列同一性。In some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders). disease), comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 25 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:25; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:35 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:35.
在一些实施例中，本文所述抗C5a抗体是包含IgG1或IgG4恒定区的全长抗C5a抗体。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region. In some embodiments, the IgG1 is human IgG1. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
例如，在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体(例如，全长抗C5a抗体)的药物组合物，重链可变结构域(VH)，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及轻链可变结构域(VL)，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。在一些实施例中，所述抗C5a抗体是全长抗体。在一些实施例中，所述全长抗C5a抗体是IgG1或IgG4抗体。在一些实施例中，所述疾病或病症选自例如，炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。在一些实施例中，所述个体是人类。For example, in some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplantation). A method for an individual subject to an immune dysregulation disease), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 12. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 20, or a variant of the V L , which contains up to about 5 LC-CDRs. Amino acid substitutions. In some embodiments, the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody. In some embodiments, the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic transplant rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers. In some embodiments, the individual is a human.
在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体的组合物，其中所述抗体包含：VH，所述VH包含SEQ ID NO:26所示的氨基酸序列或其变体，所述变体与SEQ ID NO:26所示的氨基酸序列具有至少约80％序列同一性；以及VL，所述VL包含SEQ ID NO:36所示的氨基酸序列或其变体，所述变体与SEQ ID NO:36所示的氨基酸序列具有至少约80％序列同一性。In some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders). disease), comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 26 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:26; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:36 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:36.
在一些实施例中，本文所述抗C5a抗体是包含IgG1或IgG4恒定区的全长抗C5a抗体。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region. In some embodiments, the IgG1 is human IgG1. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
例如，在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体(例如，全长抗C5a抗体)的药物组合物，重链可变结构域(VH)，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:8，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及轻链可变结构域(VL)，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:13，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:21，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。在一些实施例中，所述抗C5a抗体是全长抗体。在一些实施例中，所述全长抗C5a抗体是IgG1或IgG4抗体。在一些实施例中，所述疾病或病症选自例如，炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。在一些实施例中，所述个体是人类。For example, in some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplantation). A method for an individual suffering from an immune dysregulation disease), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 8, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 13. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 21, or a variant of the V L , which contains up to about 5 LC-CDRs. Amino acid substitutions. In some embodiments, the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody. In some embodiments, the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers. In some embodiments, the individual is a human.
在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体的组合物，其中所述抗体包含：VH，所述VH包含SEQ ID NO:27所示的氨基酸序列或其变体，所述变体与SEQ ID NO:27所示的氨基酸序列具有至少约80％序列同一性；以及VL，所述VL包含SEQ ID NO:37所示的氨基酸序列或其变体，所述变体与SEQ ID NO:37所示的氨基酸序列具有至少约80％序列同一性。In some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders). disease), comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 27 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:27; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:37 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:37.
在一些实施例中，本文所述抗C5a抗体是包含IgG1或IgG4恒定区的全长抗C5a抗体。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region. In some embodiments, the IgG1 is human IgG1. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
例如，在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体(例如，全长抗C5a抗体)的药物组合物，重链可变结构域(VH)，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及轻链可变结构域(VL)，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。在一些实施例中，所述抗C5a抗体是全长抗体。在一些实施例中，所述全长抗C5a抗体是IgG1或IgG4抗体。在一些实施例中，所述疾病或病症选自例如，炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。在一些实施例中，所述个体是人类。For example, in some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplantation). A method for an individual suffering from an immune dysregulation disease), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 14. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or a variant of the V L , which contains up to about 5 LC-CDRs. Amino acid substitutions. In some embodiments, the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody. In some embodiments, the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers. In some embodiments, the individual is a human.
在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体的组合物，其中所述抗体包含：VH，所述VH包含SEQ ID NO:28所示的氨基酸序列或其变体，所述变体与SEQ ID NO:28所示的氨基酸序列具有至少约80％序列同一性；以及VL，所述VL包含SEQ ID NO:38所示的氨基酸序列或其变体，所述变体与SEQ ID NO:38所示的氨基酸序列具有至少约80％序列同一性。In some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders). disease), comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 28 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:28; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:38 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:38.
在一些实施例中，本文所述抗C5a抗体是包含IgG1或IgG4恒定区的全长抗C5a抗体。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region. In some embodiments, the IgG1 is human IgG1. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
例如，在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体(例如，全长抗C5a抗体)的药物组合物，重链可变结构域(VH)，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:5，和HC-CDR3，其包含氨基酸序列SEQ ID NO:10，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及轻链可变结构域(VL)，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。在一些实施例中，所述抗C5a抗体是全长抗体。在一些实施例中，所述全长抗C5a抗体是IgG1或IgG4抗体。在一些实施例中，所述疾病或病症选自例如，炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。在一些实施例中，所述个体是人类。For example, in some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplantation). A method for an individual subject to an immune dysregulation disease), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 5, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 10, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 14. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or a variant of the V L , which contains up to about 5 LC-CDRs. Amino acid substitutions. In some embodiments, the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody. In some embodiments, the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic transplant rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers. In some embodiments, the individual is a human.
在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体的组合物，其中所述抗体包含：VH，所述VH包含SEQ ID NO:29所示的氨基酸序列或其变体，所述变体与SEQ ID NO:29所示的氨基酸序列具有至少约80％序列同一性；以及VL，所述VL包含SEQ ID NO:39所示的氨基酸序列或其变体，所述变体与SEQ ID NO:39所示的氨基酸序列具有至少约80％序列同一性。In some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders). disease), comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 29 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:29; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:39 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:39.
在一些实施例中，本文所述抗C5a抗体是包含IgG1或IgG4恒定区的全长抗C5a抗体。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region. In some embodiments, the IgG1 is human IgG1. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
例如，在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体(例如，全长抗C5a抗体)的药物组合物，重链可变结构域(VH)，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及轻链可变结构域(VL)，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。在一些实施例中，所述抗C5a抗体是全长抗体。在一些实施例中，所述全长抗C5a抗体是IgG1或IgG4抗体。在一些实施例中，所述疾病或病症选自例如，炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。在一些实施例中，所述个体是人类。For example, in some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplantation). A method for an individual suffering from an immune dysregulation disease), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 14. LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , which contains up to about 5 LC-CDRs Amino acid substitutions. In some embodiments, the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody. In some embodiments, the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers. In some embodiments, the individual is a human.
在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体的组合物，其中所述抗体包含：VH，所述VH包含SEQ ID NO:30所示的氨基酸序列或其变体，所述变体与SEQ ID NO:30所示的氨基酸序列具有至少约80％序列同一性；以及VL，所述VL包含SEQ ID NO:40所示的氨基酸序列或其变体，所述变体与SEQ ID NO:40所示的氨基酸序列具有至少约80％序列同一性。In some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders). disease), comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 30 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:30; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:40 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:40.
在一些实施例中，本文所述抗C5a抗体是包含IgG1或IgG4恒定区的全长抗C5a抗体。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region. In some embodiments, the IgG1 is human IgG1. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
例如，在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体(例如，全长抗C5a抗体)的药物组合物，重链可变结构域(VH)，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及轻链可变结构域(VL)，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:23，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。在一些实施例中，所述抗C5a抗体是全长抗体。在一些实施例中，所述全长抗C5a抗体是IgG1或IgG4抗体。在一些实施例中，所述疾病或病症选自例如，炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。在一些实施例中，所述个体是人类。For example, in some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplantation). A method for an individual suffering from an immune dysregulation disease), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 9, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 12. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 23, or a variant of the V L , which contains up to about 5 LC-CDRs. Amino acid substitutions. In some embodiments, the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody. In some embodiments, the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers. In some embodiments, the individual is a human.
在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体的组合物，其中所述抗体包含：VH，所述VH包含SEQ ID NO:31所示的氨基酸序列或其变体，所述变体与SEQ ID NO:31所示的氨基酸序列具有至少约80％序列同一性；以及VL，所述VL包含SEQ ID NO:41所示的氨基酸序列或其变体，所述变体与SEQ ID NO:41所示的氨基酸序列具有至少约80％序列同一性。In some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders). disease), comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 31 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:31; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:41 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:41.
在一些实施例中，本文所述抗C5a抗体是包含IgG1或IgG4恒定区的全长抗C5a抗体。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region. In some embodiments, the IgG1 is human IgG1. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
例如，在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体(例如，全长抗C5a抗体)的药物组合物，重链可变结构域(VH)，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及轻链可变结构域(VL)，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:15，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。在一些实施例中，所述抗C5a抗体是全长抗体。在一些实施例中，所述全长抗C5a抗体是IgG1或IgG4抗体。在一些实施例中，所述疾病或病症选自例如，炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。在一些实施例中，所述个体是人类。For example, in some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplantation). A method for an individual suffering from an immune dysregulation disease), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 15. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or a variant of the V L , which contains up to about 5 LC-CDRs. Amino acid substitutions. In some embodiments, the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody. In some embodiments, the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers. In some embodiments, the individual is a human.
在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体的组合物，其中所述抗体包含：VH，所述VH包含SEQ ID NO:32所示的氨基酸序列或其变体，所述变体与SEQ ID NO:32所示的氨基酸序列具有至少约80％序列同一性；以及VL，所述VL包含SEQ ID NO:42所示的氨基酸序列或其变体，所述变体与SEQ ID NO:42所示的氨基酸序列具有至少约80％序列同一性。In some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders). disease), comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 32 or Variants thereof, which variants have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:32; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:42 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:42.
在一些实施例中，本文所述抗C5a抗体是包含IgG1或IgG4恒定区的全长抗C5a抗体。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region. In some embodiments, the IgG1 is human IgG1. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
例如，在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体(例如，全长抗C5a抗体)的药物组合物，重链可变结构域(VH)，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及轻链可变结构域(VL)，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。在一些实施例中，所述抗C5a抗体是全长抗体。在一些实施例中，所述全长抗C5a抗体是IgG1或IgG4抗体。在一些实施例中，所述疾病或病症选自例如，炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。在一些实施例中，所述个体是人类。For example, in some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplantation). A method for an individual subject to an immune dysregulation disease), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 7, or said Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 14. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or a variant of the V L , which contains up to about 5 LC-CDRs. Amino acid substitutions. In some embodiments, the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody. In some embodiments, the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic transplant rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers. In some embodiments, the individual is a human.
在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体的组合物，其中所述抗体包含：VH，所述VH包含SEQ ID NO:33所示的氨基酸序列或其变体，所述变体与SEQ ID NO:33所示的氨基酸序列具有至少约80％序列同一性；以及VL，所述VL包含SEQ ID NO:43所示的氨基酸序列或其变体，所述变体与SEQ ID NO:43所示的氨基酸序列具有至少约80％序列同一性。In some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders). disease), comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 33 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:33; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:43 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:43.
在一些实施例中，本文所述抗C5a抗体是包含IgG1或IgG4恒定区的全长抗C5a抗体。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region. In some embodiments, the IgG1 is human IgG1. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
例如，在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体(例如，全长抗C5a抗体)的药物组合物，重链可变结构域(VH)，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:11，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及轻链可变结构域(VL)，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:16，LC-CDR2，其包含氨基酸序列SEQ ID NO:19，和LC-CDR3，其包含氨基酸序列SEQ ID NO:24，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。在一些实施例中，所述抗C5a抗体是全长抗体。在一些实施例中，所述全长抗C5a抗体是IgG1或IgG4抗体。在一些实施例中，所述疾病或病症选自例如，炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。在一些实施例中，所述个体是人类。For example, in some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplantation). A method for an individual suffering from an immune dysregulation disease), comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-C5a antibody (e.g., a full-length anti-C5a antibody), a heavy chain variable domain ( VH ), said V H includes: HC-CDR1, which includes the amino acid sequence SEQ ID NO: 1, HC-CDR2, which includes the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which includes the amino acid sequence SEQ ID NO: 11, or the Variants of VH comprising up to about 5 amino acid substitutions in the HC-CDRs; and light chain variable domains (VL ) comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 16. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 19, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 24, or a variant of the V L , which contains up to about 5 LC-CDRs. Amino acid substitutions. In some embodiments, the anti-C5a antibody is a full-length antibody. In some embodiments, the full-length anti-C5a antibody is an IgGl or IgG4 antibody. In some embodiments, the disease or disorder is selected from, for example, inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, acute and chronic graft rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers. In some embodiments, the individual is a human.
在一些实施例中，提供一种用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的个体的方法，包括向所述个体施用有效量的包含抗C5a抗体的组合物，其中所述抗体包含：VH，所述VH包含SEQ ID NO:34所示的氨基酸序列或其变体，所述变体与SEQ ID NO:34所示的氨基酸序列具有至少约80％序列同一性；以及VL，所述VL包含SEQ ID NO:44所示的氨基酸序列或其变体，所述变体与SEQ ID NO:44所示的氨基酸序列具有至少约80％序列同一性。In some embodiments, a method is provided for treating diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders). disease), comprising administering to the individual an effective amount of a composition comprising an anti-C5a antibody, wherein the antibody comprises: VH , the VH comprising the amino acid sequence set forth in SEQ ID NO: 34 or Variants thereof, which have at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:34; and VL , which VL comprises the amino acid sequence shown in SEQ ID NO:44 or a variant thereof A variant having at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:44.
在一些实施例中，本文所述抗C5a抗体是包含IgG1或IgG4恒定区的全长抗C5a抗体。在一些实施例中，所述IgG1是人IgG1。在一些实施例中，所述IgG4是人IgG4。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:45组成。在一些实施例中，重链恒定区包含或由氨基酸序列SEQ ID NO:46组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:47组成。在一些实施例中，轻链恒定区包含或由氨基酸序列SEQ ID NO:48组成。In some embodiments, an anti-C5a antibody described herein is a full-length anti-C5a antibody comprising an IgGl or IgG4 constant region. In some embodiments, the IgG1 is human IgG1. In some embodiments, the IgG4 is human IgG4. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO:45. In some embodiments, the heavy chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 46. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO:47. In some embodiments, the light chain constant region comprises or consists of the amino acid sequence SEQ ID NO: 48.
在一些实施例中，所述个体是哺乳动物(例如人、非人灵长类、大鼠、小鼠、牛、马、猪、绵羊、山羊、狗、猫等)。在一些实施例中，所述个体是人类。在一些实施例中，所述个体是临床患者、临床试验志愿者、实验动物等。在一些实施例中，所述个体年龄小于60岁(包括例如小于50、40、30、25、20、15或10岁)。在一些实施例中，所述个体年龄大于60岁(包括例如大于70、80、90或100岁)。在一些实施例中，所述个体是被诊断为或在遗传角度上易患本文所描述的一种或多种疾病或病症(例如自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)。在一些实施例中，所述个体具有一种或多种与本文所述的一种或多种疾病或病症相关的风险因子。In some embodiments, the subject is a mammal (eg, human, non-human primate, rat, mouse, cow, horse, porcine, sheep, goat, dog, cat, etc.). In some embodiments, the individual is a human. In some embodiments, the individual is a clinical patient, clinical trial volunteer, experimental animal, etc. In some embodiments, the individual is less than 60 years old (including, for example, less than 50, 40, 30, 25, 20, 15, or 10 years old). In some embodiments, the individual is older than 60 years old (including, for example, older than 70, 80, 90, or 100 years old). In some embodiments, the individual is diagnosed with or genetically predisposed to one or more diseases or conditions described herein (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or or pain and/or respiratory and/or transplant immune disorders). In some embodiments, the individual has one or more risk factors associated with one or more diseases or conditions described herein.
在一些实施例中，本申请提供一种向个体中在其表面表达C5a的细胞递送抗C5a抗体(例如本文所述的任一种抗C5a抗体，例如分离的抗C5a抗体)的方法，所述方法包括向该个体施用包含抗C5a抗体的组合物。In some embodiments, the application provides a method of delivering an anti-C5a antibody (e.g., any of the anti-C5a antibodies described herein, e.g., an isolated anti-C5a antibody) to cells in an individual that express C5a on their surface, said The method includes administering to the individual a composition comprising an anti-C5a antibody.
自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病或其它任何表现出C5a异常表达的疾病的许多诊断方法和这些疾病的临床描述在本领域是已知的。这类方法包括，但不限于，例如免疫组化、PCR以及荧光原位杂交(FISH)。Many diagnostic methods for autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune dysregulation diseases or any other diseases showing aberrant expression of C5a and clinical description of these diseases are known in the art. Such methods include, but are not limited to, immunohistochemistry, PCR, and fluorescence in situ hybridization (FISH), for example.
在一些实施例中，本申请所述抗C5a抗体(例如全长抗C5a抗体)和/或组合物与第二、第三或第四药剂(包括例如抗肿瘤剂、生长抑制剂、细胞毒素剂或化学治疗剂)联合使用来治疗与C5a信号通路相关的疾病。In some embodiments, the anti-C5a antibodies (e.g., full-length anti-C5a antibodies) and/or compositions described herein are combined with a second, third, or fourth agent (including, for example, an anti-tumor agent, a growth inhibitory agent, a cytotoxic agent or chemotherapeutic agents) to treat diseases related to the C5a signaling pathway.
采用例如肿瘤消退、肿瘤重量或尺寸的减少、进展时间、生存期、无进展生存期、总体响应率、响应持续时间、生存质量、蛋白表达水平和/或活性来评估癌症治疗。可以采用确定治疗效果的方法，包括例如，通过放射成像来检测是否响应。Cancer treatments are evaluated using, for example, tumor regression, reduction in tumor weight or size, time to progression, survival, progression-free survival, overall response rate, duration of response, quality of life, protein expression levels and/or activity. Methods of determining the effectiveness of treatment may be employed, including, for example, detection of response by radiographic imaging.
在一些实施例中，治疗的效果以肿瘤生长抑制百分率(％TGI)来评估，使用等式100-(T/C×100)来计算，其中T是已治疗肿瘤的相对平均肿瘤体积，C是未治疗肿瘤的相对平均肿瘤体积。在一些实施例中，％TGI为10％、20％、30％、40％、50％、60％、70％、80％、90％、91％、92％、93％、94％、95％或超过95％。在一些实施例中，治疗效果通过粒细胞形态变化和/或粒细胞存活数量的增加来评估。在一些实施例中，治疗效果通过单核细胞分泌细胞因子的增加来评估。In some embodiments, the effect of treatment is evaluated as percent tumor growth inhibition (%TGI), calculated using the equation 100 - (T/C × 100), where T is the relative mean tumor volume of the treated tumor and C is Relative mean tumor volume of untreated tumors. In some embodiments, the %TGI is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95% or more than 95%. In some embodiments, the therapeutic effect is assessed by changes in granulocyte morphology and/or an increase in the number of viable granulocytes. In some embodiments, therapeutic efficacy is assessed by increased secretion of cytokines by monocytes.
抗C5a抗体的给药剂量和方法Dosage and method of administration of anti-C5a antibodies
施用于个体(例如人)的抗C5a抗体(例如分离的抗C5a抗体)组合物的剂量可能因特定组合物、给药方式和治疗疾病类型的不同而不同。在一些实施例中，组合物(例如，包含抗C5a抗体的组合物)的量可在自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病治疗中有效产生客观响应(例如，部分响应或完全响应)。在一些实施例中，抗C5a抗体组合物的量足以在个体中产生完全响应。在一些实施例中，抗C5a抗体组合物的量足以在个体中产生部分响应。在一些实施例中，抗C5a抗体组合物的给药剂量(例如当单独施用时)足以在使用抗C5a抗体组合物治疗的个体群体中产生高于20％、25％、30％、35％、40％、45％、50％、55％、60％、64％、65％、70％、75％、80％、85％或90％的总响应率。个体对本文所述治疗方法的响应可通过，例如，RECIST的水平来确定。The dosage of an anti-C5a antibody (eg, isolated anti-C5a antibody) composition administered to an individual (eg, a human) may vary depending on the particular composition, the mode of administration, and the type of disease being treated. In some embodiments, the amount of the composition (e.g., a composition comprising an anti-C5a antibody) can be used in autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immunity. Effective in producing an objective response (eg, partial response or complete response) in the treatment of the disorder. In some embodiments, the amount of anti-C5a antibody composition is sufficient to produce a complete response in an individual. In some embodiments, the amount of anti-C5a antibody composition is sufficient to produce a partial response in an individual. In some embodiments, the anti-C5a antibody composition is administered at a dose (e.g., when administered alone) sufficient to produce greater than 20%, 25%, 30%, 35%, or more in a population of individuals treated with the anti-C5a antibody composition. Overall response rate of 40%, 45%, 50%, 55%, 60%, 64%, 65%, 70%, 75%, 80%, 85%, or 90%. An individual's response to the treatment methods described herein can be determined, for example, by the level of RECIST.
在一些实施例中，组合物(例如包含分离的抗C5a抗体的组合物)的量足以延长个体的无进展生存期。在一些实施例中，组合物的量足以延长个体的总体生存期。在一些实施例中，在使用抗C5a抗体组合物治疗的个体群体中，组合物的量(例如当单独施用时)足以产生高于50％、60％、70％或77％的临床益处。In some embodiments, the amount of the composition (eg, a composition comprising an isolated anti-C5a antibody) is sufficient to prolong progression-free survival in an individual. In some embodiments, the amount of the composition is sufficient to prolong the overall survival of the subject. In some embodiments, the amount of the composition (eg, when administered alone) is sufficient to produce greater than 50%, 60%, 70%, or 77% of clinical benefit in a population of individuals treated with an anti-C5a antibody composition.
在一些实施例中，组合物(例如包含分离的抗C5a抗体的组合物)的量，单独使用或与第二，第三、和/或第四药剂联合使用时，是指在治疗前或与未接受治疗的其他受试者的相应活性相比，足以控制症状和减少病情加重的风险的量。可以采用标准方法来测量该疗效的大小，例如纯化酶的体外检测、基于细胞的检测、动物模型或人体试验。In some embodiments, the amount of a composition (eg, a composition comprising an isolated anti-C5a antibody), alone or in combination with a second, third, and/or fourth agent, is before treatment or with An amount sufficient to control symptoms and reduce the risk of exacerbation compared to the corresponding activity in other subjects not receiving treatment. The magnitude of this effect can be measured using standard methods, such as in vitro assays of purified enzymes, cell-based assays, animal models, or human trials.
在一些实施例中，当将组合物施用于个体时，组合物中抗C5a抗体(例如全长的抗C5a抗体)的量低于引起毒性效应(即，一种高于临床可接受毒性水平的效应)的水平，或者处于潜在副作用可以控制或耐受的水平。In some embodiments, the amount of anti-C5a antibody (e.g., full-length anti-C5a antibody) in the composition is less than the amount that causes a toxic effect (i.e., an amount that is greater than a clinically acceptable level of toxicity) when the composition is administered to an individual. effect), or at a level at which potential side effects can be controlled or tolerated.
在一些实施例中，遵循相同的给药方案，组合物的量接近的组合物的最大耐受剂量(MTD)。在一些实施例中，组合物的量高于MTD的80％、90％、95％或98％。In some embodiments, following the same dosing regimen, the amount of the composition approximates the maximum tolerated dose (MTD) of the composition. In some embodiments, the amount of the composition is greater than 80%, 90%, 95%, or 98% of the MTD.
在一些实施例中，组合物中抗C5a抗体(例如全长的抗C5a抗体)的含量在0.001μg到1000μg的范围之内。In some embodiments, the amount of anti-C5a antibody (eg, full-length anti-C5a antibody) in the composition ranges from 0.001 μg to 1000 μg.
在如上所述任一个实施例中，组合物中的抗C5a抗体(例如全长的抗C5a抗体)的有效量，按照体重时计算，为0.1μg/kg到100mg/kg的范围之内。In any of the embodiments described above, the effective amount of anti-C5a antibody (eg, full-length anti-C5a antibody) in the composition ranges from 0.1 μg/kg to 100 mg/kg based on body weight.
抗C5a抗体组合物可通过多种途径施用于个体(如人类)，包括，例如静脉注射、动脉内给药、腹腔注射、肺内给药、口服给药、吸入给药、血管内给药、肌肉注射、气管内给药、皮下注射、眼内给药、鞘内给药、粘膜给药或经皮给药。在一些实施例中，使用组合物的缓释制剂。在一些实施例中，组合物通过静脉给药。在一些实施例中，组合物通过动脉给药。在一些实施例中，组合物通过腹膜内给药。在一些实施例中，组合物通过肝内给药。在一些实施例中，组合物通过肝动脉输注给药。在一些实施例中，组合物施用于远离第一病灶的部位。Anti-C5a antibody compositions can be administered to an individual (e.g., a human) by a variety of routes, including, for example, intravenous injection, intraarterial administration, intraperitoneal injection, intrapulmonary administration, oral administration, inhalation administration, intravascular administration, Intramuscular, intratracheal, subcutaneous, intraocular, intrathecal, mucosal or transdermal administration. In some embodiments, a sustained release formulation of the composition is used. In some embodiments, the composition is administered intravenously. In some embodiments, the composition is administered intraarterially. In some embodiments, the composition is administered intraperitoneally. In some embodiments, the composition is administered intrahepatically. In some embodiments, the composition is administered by hepatic artery infusion. In some embodiments, the composition is administered to a site distal to the first lesion.
制品及试剂盒Products and kits
在本申请的一些实施例中，提供一种制品，所述制品包含一种物质，所述物质能够用于治疗与C5a信号通路相关的疾病(例如，自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)，或者用于递送抗C5a抗体(例如一种全长抗C5a抗体)到表面表达C5a的细胞。所述制品可以包括一种容器以及在容器上或随该容器附带的标签或包装说明书。合适的容器包括，例如瓶子、小瓶、注射器等。容器可以由多种材料制成，例如玻璃或塑料。通常，该容器内装有能够有效治疗本文所述疾病或病症的组合物，并且具有一个无菌端口(例如该容器可以是一个静脉输液袋或是一个具有皮下注射针头可刺穿盖子的小瓶)。组合物中的至少一种活性物质即为本申请所述的抗C5a抗体。标签或包装说明书标示了该组合物可以用于治疗的特定病症。标签或包装说明书进一步包含给患者施用抗C5a抗体组合物的说明书。包括联合治疗的制品和试剂盒均在本文的考虑范围之内。In some embodiments of the present application, there is provided an article of manufacture comprising a substance that can be used to treat diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory disorders and or cancer and/or pain and/or respiratory and/or transplant immune disorders), or for delivering an anti-C5a antibody (e.g., a full-length anti-C5a antibody) to cells expressing C5a on their surface. The article of manufacture may include a container and a label or package insert on or accompanying the container. Suitable containers include, for example, bottles, vials, syringes, and the like. Containers can be made from a variety of materials, such as glass or plastic. Typically, the container contains a composition effective for treating the disease or condition described herein and has a sterile port (eg, the container may be an IV bag or a vial with a hypodermic needle-pierceable cap). At least one active substance in the composition is the anti-C5a antibody described in this application. The label or package insert identifies the specific condition for which the composition is used to treat. The label or package insert further contains instructions for administering the anti-C5a antibody composition to a patient. Articles and kits including combination therapies are considered within the scope of this article.
包装说明书是指通常包含在治疗产品的商业包装内的说明书，其包含关于与这些治疗产品使用有关的适应症、用法、剂量、施用、禁忌症和/或警告信息。在一些实施例中，包装说明书标明该组合物可以用于治疗与C5a信号通路相关的疾病(例如自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)。在一些实施例中，包装说明书标明该组合物可以用于治疗以下的疾病，包括炎症反应综合症(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、肾功能衰竭的实体、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、肿瘤生长和实体器官癌症。Package insert means the instructions usually included in the commercial packaging of therapeutic products that contain information on indications, usage, dosage, administration, contraindications and/or warnings relevant to the use of these therapeutic products. In some embodiments, the package insert indicates that the composition can be used to treat diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or Transplant immune disorders). In some embodiments, the package insert states that the composition can be used to treat conditions including inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, Pneumonia, acute and chronic graft rejection in transplant patients, graft-versus-host reaction, glomerular disease, glomerulonephritis, entities of renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases , inflammatory bowel disease, Crohn's disease, tumor growth, and solid organ cancers.
此外，所述制品还可以包括第二容器，其包含药学上可接受的缓冲液，例如抑菌性注射用水(BWFI)、磷酸盐缓冲液、格林氏溶液或葡萄糖溶液。还可以包括从商业和用户角度而言所需的其他材料，包括其他缓冲液、稀释液、过滤器、针头和注射器。Additionally, the article of manufacture may further include a second container containing a pharmaceutically acceptable buffer, such as bacteriostatic water for injection (BWFI), phosphate buffer, Green's solution, or dextrose solution. Other materials required from a commercial and user perspective may also be included, including additional buffers, diluents, filters, needles, and syringes.
同时还涉及可用于各种目的的试剂盒，例如用于治疗与C5a信号通路相关的疾病(例如自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)，或者用于递送抗C5a抗体(例如全长抗C5a抗体)到表面表达C5a的细胞中，任选与制品组合。本申请的试剂盒包括一个或多个容器，其包含抗C5a抗体组合物(或单剂量形式和/或制品)，并且在一些实施例中，进一步包含另一种药剂(例如本文所述的药剂)和/或与本文所述任一方法相一致的使用说明书。该试剂盒可进一步包括选择适合治疗个体的描述。本申请中试剂盒中所附带的使用说明书通常是标签或包装说明书上的书面说明(例如包含在试剂盒内的纸页)，机器可读的说明(例如，磁性或光学储存光盘上的说明)也是可以接受的。Also contemplated are kits that can be used for various purposes, for example for the treatment of diseases associated with the C5a signaling pathway (e.g. autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or immune disorders), or for delivering anti-C5a antibodies (eg, full-length anti-C5a antibodies) to cells expressing C5a on their surface, optionally in combination with an article of manufacture. Kits of the present application include one or more containers comprising an anti-C5a antibody composition (or single dose form and/or article of manufacture) and, in some embodiments, further comprising another agent (e.g., an agent described herein ) and/or instructions for use consistent with any method described herein. The kit may further include instructions for selecting individuals suitable for treatment. Instructions for use accompanying the kit in this application are usually written instructions on the label or package insert (e.g., a sheet of paper included in the kit), machine-readable instructions (e.g., instructions on a magnetic or optical storage disc) Also acceptable.
例如，在一些实施例中，试剂盒包括一种包含抗C5a抗体(例如全长的抗C5a抗体)的组合物。在一些实施例中，试剂盒包括：a)包含本文所述的任一种抗C5a抗体的组合物，和b)至少一种有效量的其它药剂，其能够增强抗C5a抗体的效果(如治疗效果、检测效果)。在一些实施例中，试剂盒包括：a)包含本文所述的任一种抗C5a抗体的组合物，和b)向个体施用抗C5a抗体组合物用于治疗与C5a信号通路相关的疾病(例如自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的使用说明书。在一些实施例中，试剂盒包括：a)包含本文所述的任一种抗C5a抗体的组合物，和b)至少一种有效量的其它药剂，其能够增强抗C5a抗体的效果(如治疗效果、检测效果)和c)向个体施用抗C5a抗体组合物和其它物质用于治疗与C5a信号通路相关的疾病(例如自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)的使用说明书。所述抗C5a抗体和其他物质可以存在于独立的容器或同一个容器中。例如，该试剂盒可以包括一种特定组合物或两种或更多种组合物，其中一种组合物包括抗C5a抗体，另一种组合物包括另一种药剂。For example, in some embodiments, the kit includes a composition comprising an anti-C5a antibody (eg, a full-length anti-C5a antibody). In some embodiments, a kit includes: a) a composition comprising any one of the anti-C5a antibodies described herein, and b) an effective amount of at least one other agent capable of enhancing the effects of the anti-C5a antibody (e.g., treating effect, detection effect). In some embodiments, a kit includes: a) a composition comprising any one of the anti-C5a antibodies described herein, and b) administering the anti-C5a antibody composition to an individual for treating a disease associated with the C5a signaling pathway (e.g., Instructions for use in autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and/or respiratory and/or transplant immune disorders). In some embodiments, a kit includes: a) a composition comprising any one of the anti-C5a antibodies described herein, and b) an effective amount of at least one other agent capable of enhancing the effects of the anti-C5a antibody (e.g., treating effect, detecting effect) and c) administering anti-C5a antibody compositions and other substances to an individual for the treatment of diseases associated with the C5a signaling pathway (e.g., autoimmune diseases and/or inflammatory conditions and/or cancer and/or pain and /or respiratory and/or transplant immune disorders) instructions for use. The anti-C5a antibody and other substances may be present in separate containers or in the same container. For example, the kit may include one specific composition or two or more compositions, one of which includes an anti-C5a antibody and another of which includes another agent.
在一些实施例中，试剂盒包含一种(或一组)编码抗C5a抗体(例如全长的抗C5a抗体)的核酸。在一些实施例中，试剂盒包含：a)一种(或一组)编码抗C5a抗体(例如全长的抗C5a抗体)的核酸，和b)一种表达核酸(或一组核酸)的宿主细胞。在一些实施例中，试剂盒包含：a)一种(或一组)编码抗C5a抗体(例如全长的抗C5a抗体)的核酸，和b)使用说明书，适用于：i)在宿主细胞中表达抗C5a抗体，ii)制备包含抗C5a抗体的组合物，和iii)向个体施用包含抗C5a抗体的组合物来治疗与C5a信号通路相关的疾病(例如自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)。在一些实施例中，试剂盒包括：a)一种(或一组)编码抗C5a抗体(例如全长的抗C5a抗体)的核酸，b)一种表达核酸(或一组核酸)的宿主细胞，和c)使用说明书，适用于：i)在宿主细胞中表达抗C5a抗体，ii)制备包含抗C5a抗体的组合物，和iii)向个体施用包含抗C5a抗体的组合物来治疗与C5a信号通路相关的疾病(例如自身免疫性疾病和/或炎性病症和/或癌症和/或疼痛和/或呼吸和/或移植免疫失调性疾病)。In some embodiments, the kit includes a nucleic acid (or a set of nucleic acids) encoding an anti-C5a antibody (eg, a full-length anti-C5a antibody). In some embodiments, the kit comprises: a) a nucleic acid (or set of nucleic acids) encoding an anti-C5a antibody (eg, a full-length anti-C5a antibody), and b) a host expressing the nucleic acid (or set of nucleic acids) cell. In some embodiments, the kit comprises: a) a nucleic acid (or a set of) nucleic acids encoding an anti-C5a antibody (e.g., a full-length anti-C5a antibody), and b) instructions for: i) in a host cell Expressing an anti-C5a antibody, ii) preparing a composition comprising the anti-C5a antibody, and iii) administering the composition comprising the anti-C5a antibody to an individual to treat a disease associated with the C5a signaling pathway (e.g., autoimmune disease and/or inflammatory disorder and/or cancer and/or pain and/or respiratory and/or transplant immune disorders). In some embodiments, the kit includes: a) a nucleic acid (or a set of nucleic acids) encoding an anti-C5a antibody (eg, a full-length anti-C5a antibody), b) a host cell expressing the nucleic acid (or a set of nucleic acids) , and c) instructions for use in: i) expressing an anti-C5a antibody in a host cell, ii) preparing a composition comprising an anti-C5a antibody, and iii) administering a composition comprising an anti-C5a antibody to an individual to treat patients with C5a signaling Pathway related diseases (eg autoimmune diseases and/or inflammatory disorders and/or cancer and/or pain and/or respiratory and/or transplant immune disorders).
在一些实施例中，本申请还涉及用于检测和/或诊断的试剂盒，所述试剂盒包含本文中所述的任一种抗C5a抗体。在一些实施例中，试剂盒包含一种偶联物，所述偶联物包含本文中所述的任一种抗C5a抗体以及偶联部分，其中偶联部分为放射性同位素、荧光物质、发光物质、有色物质或酶。In some embodiments, the present application also relates to kits for detection and/or diagnosis, said kits comprising any of the anti-C5a antibodies described herein. In some embodiments, the kit includes a conjugate comprising any anti-C5a antibody described herein and a coupling moiety, wherein the coupling moiety is a radioactive isotope, a fluorescent substance, a luminescent substance , colored substances or enzymes.
在一些实施例中，涉及本申请中所述的任一种抗C5a抗体或本发明的偶联物在制备试剂盒中的用途，所述试剂盒用于检测样品中C5a的存在或其水平。In some embodiments, it relates to the use of any of the anti-C5a antibodies described in this application or the conjugate of the invention in preparing a kit for detecting the presence or level of C5a in a sample.
在一些实施例中，提供一种检测样品中C5a存在或水平的体外方法，所述方法包括：a)在允许抗体和C5a之间形成复合物的条件下将样品与本文中所述的任一种抗C5a抗体或偶联物相接触；和b)检测复合物的形成；和c)确定样品中C5a的存在或其水平。In some embodiments, an in vitro method of detecting the presence or level of C5a in a sample is provided, the method comprising: a) contacting the sample with any of the methods described herein under conditions that allow the formation of a complex between the antibody and C5a. contacting an anti-C5a antibody or conjugate; and b) detecting the formation of a complex; and c) determining the presence or level of C5a in the sample.
在一些实施例中，提供一种诊断C5a水平异常的方法，所述方法包括：a)获得待测样品，b)在允许抗体和C5a之间形成复合物的条件下将待测样品与本文中所述的任一种抗C5a抗体或偶联物相接触；c)检测复合物的形成；d)确定待测样品中C5a的水平；和e)判断待测样品中C5a的水平是否高于正常组样品中C5a的水平。In some embodiments, a method for diagnosing abnormalities in C5a levels is provided, the method comprising: a) obtaining a sample to be tested, and b) combining the sample to be tested with the method described herein under conditions that allow the formation of a complex between the antibody and C5a. Contact any of the anti-C5a antibodies or conjugates; c) detect the formation of the complex; d) determine the level of C5a in the sample to be tested; and e) determine whether the level of C5a in the sample to be tested is higher than normal Levels of C5a in group samples.
在一些实施例中，所述样品为血清样品或血浆样品。在另外一些实施例中，所述待测样品来源于有需要的患者。在进一步的优选实施例中，所述患者接受过或未接受过C5a抗体治疗。In some embodiments, the sample is a serum sample or plasma sample. In other embodiments, the sample to be tested is derived from a patient in need. In a further preferred embodiment, the patient has or has not received C5a antibody treatment.
本申请所述的试剂盒以合适的形式进行包装。合适的包装包括，但不限于，小瓶、瓶子、广口瓶、软包装(例如密封的聚酯薄膜或塑料袋)等。试剂盒可以任选地提供其它的组分，例如缓冲液和说明信息。因此，本申请还提供制品，其包括小瓶、瓶子、广口瓶、软包装(例如密封的聚酯薄膜或塑料袋)等。The kits described in this application are packaged in a suitable form. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (eg, sealed mylar or plastic bags), and the like. Kits may optionally provide other components such as buffers and instructions. Accordingly, the present application also provides articles including vials, bottles, jars, flexible packaging (eg, sealed Mylar or plastic bags), and the like.
关于抗C5a抗体组合物的使用说明书，通常包括一些信息，诸如，剂量，给药周期和给药途径等。容器可以是单位剂量的，大包装的(如，多剂量包装)或亚单位剂量的。例如，提供一种包含足够剂量的如本文所述的抗C5a抗体(例如全长的抗C5a抗体)的试剂盒以对个体进行长期有效的治疗，例如一周、8天、9天、10天、11天、12天、13天、2周、3周、4周、6周、8周、3个月、4个月、5个月、7个月、8个月、9个月或更长时间。试剂盒还可包含多单位剂量的抗C5a抗体、药物组合物和使用说明书，并且以足够在药房中储存和使用的量进行包装，例如，医院药房和复方药房。Instructions for use of anti-C5a antibody compositions generally include information such as dosage, administration period and administration route. Containers may be unit dose, bulk (eg, multi-dose packaging) or subunit dose. For example, a kit is provided that contains a sufficient dose of an anti-C5a antibody as described herein (e.g., a full-length anti-C5a antibody) to effectively treat an individual for a long period of time, such as one week, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months or longer time. The kit may also contain multiple unit doses of the anti-C5a antibody, the pharmaceutical composition and instructions for use, and be packaged in quantities sufficient for storage and use in pharmacies, such as hospital pharmacies and compounding pharmacies.
本领域的技术人员将认识到在本申请的范围和宗旨内可能的若干实施例。现在将通过参考以下非限制性实施例来更详细地描述本申请。以下实施例进一步阐明本申请，但不应解释为以任何方式进行限制其范围。Those skilled in the art will recognize that several embodiments are possible within the scope and spirit of this application. The application will now be described in more detail with reference to the following non-limiting examples. The following examples further illustrate this application but should not be construed as limiting its scope in any way.
具体实施方式Detailed ways
在下述公开的实施例中，适用如下缩写：补体成分5a(complement component 5a，C5a)；Bavih-C5a(Biotin-Avi-10his-C5a)In the following disclosed embodiments, the following abbreviations apply: complement component 5a (complement component 5a, C5a); Bavih-C5a (Biotin-Avi-10his-C5a)
实施例1：制备重组人C5a并筛选抗C5a的单链抗体(scFv)Example 1: Preparation of recombinant human C5a and screening of anti-C5a single chain antibodies (scFv)
1.1制备重组C5a抗原1.1 Preparation of recombinant C5a antigen
通过亚克隆将编码人C5a的cDNA(由上海捷瑞生物工程有限公司合成)构建到原核表达载体pTWIN1或真核表达载体pTT5中。在编码上述蛋白的cDNA的末端添加His标签或其它的本领域技术人员常用标签。构建产生pTWIN1-C5a和pTT5-Avi-10his-C5a表达载体。其中“his或His代表His标签，“Avi”代表亲和素标签。The cDNA encoding human C5a (synthesized by Shanghai Jierui Bioengineering Co., Ltd.) was constructed into the prokaryotic expression vector pTWIN1 or the eukaryotic expression vector pTT5 by subcloning. Add His tag or other tags commonly used by those skilled in the art at the end of the cDNA encoding the above protein. The pTWIN1-C5a and pTT5-Avi-10his-C5a expression vectors were constructed and produced. Among them, "his" or "His" represents the His tag, and "Avi" represents the avidin tag.
按照仪器厂商及试剂盒操作说明书，对重组人Avi-10his-C5a进行表达和纯化。简言之，将真核表达载体pTT5-Avi-10his-C5a转染到293F细胞中，并将上述细胞在37℃、8％CO2、120rpm条件下培养5天。收集细胞培养液。根据操作说明书，采用镍柱(Ni)纯化表达His标签的蛋白。具体操作如下：采用QIAGEN公司的Ni-NTA进行固定化金属亲和层析(IMAC)。首先采用缓冲液A1(50mM Na3PO4、0.15M NaCl、pH 7.2)平衡镍柱，流速150cm/h。将培养液上清的pH调整至7.2，室温上样，流速150cm/h。随后，采用6倍柱体积的A1缓冲液再次平衡该柱，流速150cm/h。最后，采用10倍柱体积的50mM PB溶液(包含0.15M NaCl和0.2M咪唑，pH 7.2)进行洗脱，收集洗脱液。Express and purify recombinant human Avi-10his-C5a according to the instrument manufacturer and kit operating instructions. Briefly, the eukaryotic expression vector pTT5-Avi-10his-C5a was transfected into 293F cells, and the above cells were cultured at 37°C, 8% CO 2 , and 120 rpm for 5 days. Collect cell culture fluid. According to the operating instructions, a nickel column (Ni) was used to purify the protein expressing His tag. The specific operation is as follows: Immobilized metal affinity chromatography (IMAC) is performed using Ni-NTA from QIAGEN Company. First, use buffer A1 (50mM Na 3 PO 4 , 0.15M NaCl, pH 7.2) to balance the nickel column, with a flow rate of 150cm/h. The pH of the culture supernatant was adjusted to 7.2, and the sample was loaded at room temperature with a flow rate of 150cm/h. Subsequently, the column was equilibrated again with 6 column volumes of A1 buffer at a flow rate of 150 cm/h. Finally, 10 times the column volume of 50mM PB solution (containing 0.15M NaCl and 0.2M imidazole, pH 7.2) was used for elution, and the eluent was collected.
按照仪器厂商及试剂盒操作说明书，对重组人C5a进行表达和纯化。简言之，将重组人C5a原核表达载体pTWIN1-C5a转化大肠杆菌并进行扩大培养。收集菌体后，加入300ml 50mM Tris溶液，搅拌溶解，放入-40℃冰箱过夜。第二天，将上述样品取出融解后，使用超声破碎仪破碎细胞。离心后取上清，用0.8um滤膜过滤上清。加水将滤液稀释至1L，并用稀释10倍的冰醋酸将其pH调整至6.5，再用0.45um滤膜过滤。样品处理完成后依次进行离子交换层析(CM-琼脂糖凝胶FF)、凝胶过滤层析(Superdex 75)和离子交换柱层析(SPHP)纯化，收集洗脱产物浓缩后测定浓度，备用。Express and purify recombinant human C5a according to the instrument manufacturer and kit operating instructions. Briefly, the recombinant human C5a prokaryotic expression vector pTWIN1-C5a was transformed into E. coli and expanded cultured. After collecting the bacterial cells, add 300ml of 50mM Tris solution, stir to dissolve, and place in a -40°C refrigerator overnight. The next day, the above samples were taken out and thawed, and the cells were disrupted using an ultrasonic disruptor. After centrifugation, take the supernatant and filter the supernatant with a 0.8um filter membrane. Add water to dilute the filtrate to 1L, adjust its pH to 6.5 with glacial acetic acid diluted 10 times, and then filter with a 0.45um filter membrane. After the sample processing is completed, purify by ion exchange chromatography (CM-Sepharose FF), gel filtration chromatography (Superdex 75) and ion exchange column chromatography (SPHP). Collect the eluted products and concentrate them to determine the concentration for later use. .
1.2制备生物素化标记的C5a抗原1.2 Preparation of biotinylated C5a antigen
按照操作说明书，采用生物素化连接酶B0101A(GeneCopoeia)对Avi-10His-C5a进行生物素化标记。简言之，向Avi-10His-C5a中加入BufferA/B和BirA连接酶后在30℃下孵育2小时。生物素化的Avi-10His-C5a被命名为Bavih-C5a。通过ELISA方法检测生物素化效率。简言之，将Bavih-C5a起始浓度设置为500ng/ml，按1:2的比例倍比稀释，稀释之后包被ELISA板。采用SA-HRP来检测信号，用生物素化标准品做对照。确定Bavih-C5a生物素化标记效率为70％。According to the operating instructions, Avi-10His-C5a was biotinylated using biotinylated ligase B0101A (GeneCopoeia). Briefly, BufferA/B and BirA ligase were added to Avi-10His-C5a and incubated at 30°C for 2 hours. Biotinylated Avi-10His-C5a was named Bavih-C5a. Biotinylation efficiency was detected by ELISA method. Briefly, the starting concentration of Bavih-C5a was set to 500ng/ml, diluted at a ratio of 1:2, and then coated on the ELISA plate after dilution. SA-HRP was used to detect the signal, and biotinylated standards were used as controls. The Bavih-C5a biotinylation labeling efficiency was determined to be 70%.
1.3筛选抗C5a单链抗体(scFv)1.3 Screening anti-C5a single chain antibodies (scFv)
构建scFv抗体噬菌体展示文库：使用重组人C5a作为抗原，和佐剂一起免疫家兔，取免疫后的动物血清，ELISA检测免疫后家兔血清中总IgG滴度。几轮免疫后，使用外周血、淋巴结和脾脏，建立噬菌体展示库。简言之，取免疫后的家兔的外周血、淋巴结和脾脏，并提取RNA，经逆转录获得cDNA。采用VH和VK特异性引物扩增VH和VK片段，经胶回收纯化后，连接VH和VK，构建scFv，并将其克隆到噬菌体展示质粒pDAN5中。随后，将该质粒电转至大肠杆菌TG1中，利用噬菌体感染大肠杆菌TG1，获得scFv抗体噬菌体展示文库。Construct scFv antibody phage display library: use recombinant human C5a as the antigen, immunize rabbits together with adjuvant, collect the animal serum after immunization, and detect the total IgG titer in the rabbit serum after immunization by ELISA. After several rounds of immunization, peripheral blood, lymph nodes, and spleen were used to establish a phage display library. Briefly, peripheral blood, lymph nodes and spleens of immunized rabbits were collected, RNA was extracted, and cDNA was obtained by reverse transcription. VH and VK fragments were amplified using VH and VK specific primers. After gel recovery and purification, VH and VK were ligated to construct scFv and cloned into the phage display plasmid pDAN5. Subsequently, the plasmid was electroporated into E. coli TG1, and the phage was used to infect E. coli TG1 to obtain a scFv antibody phage display library.
筛选抗C5a单链抗体(scFv)：经过一系列重复筛选步骤，从噬菌体展示库中分离获得特异性结合人C5a的scFv抗体。简言之，取2x1011PFU的噬菌体scFv文库加入到Bavih-C5a中，37℃孵育2小时。结合人C5a的噬菌体被包被了链霉亲和素的磁珠捕获，而未结合的噬菌体被洗掉。采用TBST溶液洗涤8-15遍(随着筛选轮数增加，洗涤次数增加)之后，用Glycine-HCl溶液(pH2.2)洗脱特异性结合人C5a的噬菌体。用这些噬菌体感染指数生长期的TG1细胞，加入氨苄西林培养1小时后，加入辅助噬菌体，28℃、200rpm摇床培养过夜。次日收集培养液，离心后获得上清，进入下一轮筛选，直至获得阳性scFv抗体文库。Screening of anti-C5a single chain antibodies (scFv): After a series of repeated screening steps, scFv antibodies that specifically bind to human C5a are isolated from the phage display library. Briefly, 2x10 11 PFU of the phage scFv library was added to Bavih-C5a and incubated at 37°C for 2 hours. Phage that bind to human C5a are captured by streptavidin-coated magnetic beads, while unbound phage are washed away. After washing 8-15 times with TBST solution (as the number of screening rounds increases, the number of washes increases), Glycine-HCl solution (pH 2.2) is used to elute the phage that specifically binds human C5a. Use these phages to infect TG1 cells in the exponential growth phase, add ampicillin and culture for 1 hour, add helper phages, and culture overnight at 28°C and 200rpm shaker. The culture fluid was collected the next day, centrifuged to obtain the supernatant, and entered into the next round of screening until a positive scFv antibody library was obtained.
采用ELISA筛选C5a单链抗体(scFv)：将MACS淘选后富集的噬菌体进行ELISA筛选。简言之，将重组人C5a抗原溶解在PBS溶液中，按照0.1μg/孔包被96孔板，4℃过夜。在加入抗体之前，用PBST溶液洗涤96孔板。每孔中加入90μL含有4％脱脂奶粉的PBS，随后向对应的孔中加入10μL phage-scFv表达上清，37℃孵育1-2小时。用PBST溶液洗涤后，加入HRP标记的抗M13单抗(Sino Biological，11973-MM05T-H)，100μL/孔，37℃孵育1h。PBST洗板3次，加TMB显色液，100μL/孔，室温避光孵育10min。用2M H2SO4终止显色反应，用酶标仪读取450nm处的吸光度值。挑选阳性克隆菌进行测序，测序正确的阳性克隆菌扩大培养后，提取噬菌体展示质粒，备用。在筛选过程结束后，获得一系列阳性scFv抗体。Use ELISA to screen C5a single chain antibodies (scFv): The phages enriched after MACS panning were screened by ELISA. Briefly, recombinant human C5a antigen was dissolved in PBS solution, coated on a 96-well plate at 0.1 μg/well, and incubated at 4°C overnight. Before adding the antibody, wash the 96-well plate with PBST solution. Add 90 μL of PBS containing 4% skim milk powder to each well, then add 10 μL of phage-scFv expression supernatant to the corresponding well, and incubate at 37°C for 1-2 hours. After washing with PBST solution, add HRP-labeled anti-M13 monoclonal antibody (Sino Biological, 11973-MM05T-H), 100 μL/well, and incubate at 37°C for 1 hour. Wash the plate three times with PBST, add TMB chromogenic solution, 100 μL/well, and incubate at room temperature in the dark for 10 minutes. The color reaction was terminated with 2M H 2 SO 4 , and the absorbance value at 450 nm was read with a microplate reader. Select the positive clones for sequencing. After the correctly sequenced positive clones are expanded and cultured, the phage display plasmid is extracted and used for later use. At the end of the screening process, a series of positive scFv antibodies were obtained.
实施例2：制备和表征全长抗C5a嵌合抗体Example 2: Preparation and characterization of full-length anti-C5a chimeric antibodies
2.1制备全长抗C5a嵌合抗体2.1 Preparation of full-length anti-C5a chimeric antibodies
将获得的阳性scFv抗体重构成具有人IgG1或IgG4的重链恒定区和人kappa或lambda轻链恒定区的人IgG1或IgG4抗体分子。从噬菌体表达载体中扩增VL和VH，分别构建入真核表达载体pTT5-K(包含kappa恒定区)或pTT5-L(包含lambda恒定区)和pTT5-H1(包含IgG1重链恒定区)或pTT5-H4(包含IgG4重链恒定区)中。将表达轻链的质粒和表达重链的质粒共转染293F细胞。37℃，8％CO2，120rpm培养5天，用Protein A亲和层析柱纯化培养液。简言之，首先采用6倍柱体积的PBS缓冲液(包含50mM PBS和0.15M NaCl，pH7.2)以150cm/h的流速平衡蛋白A柱。将培养液上清的pH调整至7.2，室温上样，流速150cm/h。至完全平衡后，加入50mM的柠檬酸钠缓冲液(pH3.5)进行洗脱，收集洗脱液。The obtained positive scFv antibody is reconstituted into a human IgG1 or IgG4 antibody molecule having a heavy chain constant region of human IgG1 or IgG4 and a human kappa or lambda light chain constant region. V L and V H were amplified from the phage expression vector and constructed into the eukaryotic expression vector pTT5-K (containing the kappa constant region) or pTT5-L (containing the lambda constant region) and pTT5-H1 (containing the IgG1 heavy chain constant region) respectively. ) or pTT5-H4 (containing the IgG4 heavy chain constant region). The plasmid expressing the light chain and the plasmid expressing the heavy chain were co-transfected into 293F cells. Cultivate for 5 days at 37°C, 8% CO 2 , 120 rpm, and purify the culture solution using a Protein A affinity chromatography column. Briefly, the Protein A column was first equilibrated with 6 column volumes of PBS buffer (containing 50 mM PBS and 0.15 M NaCl, pH 7.2) at a flow rate of 150 cm/h. The pH of the culture supernatant was adjusted to 7.2, and the sample was loaded at room temperature with a flow rate of 150cm/h. After complete equilibrium, 50 mM sodium citrate buffer (pH 3.5) was added for elution, and the eluate was collected.
2.2抗C5a抗体的亲和力2.2 Affinity of anti-C5a antibodies
C5a ELISA结合实验：C5a ELISA binding experiment:
采用ELISA结合实验评价全长抗C5a抗体RH23、RH25、RH32、RH35、RH36、RH58、RH85、RH86、RH92、RH95(重构建成人IgG4形式)与人C5a的亲和力，并以INab308作为对照。简言之，将实施例1制备的重组人C5a抗原溶解在PBS溶液中，按照0.1μg/孔包被96孔板，4℃过夜。在加入抗体之前，用TBST溶液洗涤96孔板，用5％牛奶37℃封闭1-2小时，再用TBST溶液洗涤。首先将每个抗体样品稀释至10μg/mL，随后以1：3比例进行梯度稀释。将梯度稀释后的样品分别加入96孔板中，每孔100μL，37℃孵育1小时。随后用TBST溶液洗涤6次，加入羊抗人IgG Fc-AP抗体(SouthernBiotech，2048-04；1：2500)，每孔100μL，37℃孵育1小时。TBST洗板机洗涤6次后，每孔加入pNPP 50μL，37℃孵育10min，用3M NaOH终止反应。读取410nm处的吸光值，并通过PRISM生成结合曲线。An ELISA binding experiment was used to evaluate the affinity of full-length anti-C5a antibodies RH23, RH25, RH32, RH35, RH36, RH58, RH85, RH86, RH92, and RH95 (reconstructed into human IgG4 form) with human C5a, and INab308 was used as a control. Briefly, the recombinant human C5a antigen prepared in Example 1 was dissolved in PBS solution, coated on a 96-well plate at 0.1 μg/well, and incubated at 4°C overnight. Before adding the antibody, wash the 96-well plate with TBST solution, block with 5% milk at 37°C for 1-2 hours, and then wash with TBST solution. Each antibody sample was first diluted to 10 μg/mL, followed by gradient dilution in a 1:3 ratio. Add the gradient diluted samples to a 96-well plate, 100 μL per well, and incubate at 37°C for 1 hour. Then wash 6 times with TBST solution, add goat anti-human IgG Fc-AP antibody (SouthernBiotech, 2048-04; 1:2500), 100 μL per well, and incubate at 37°C for 1 hour. After washing with TBST plate washer 6 times, add 50 μL of pNPP to each well, incubate at 37°C for 10 min, and terminate the reaction with 3M NaOH. The absorbance value at 410 nm was read and a binding curve was generated by PRISM.
结果如图1A-1C所示，与对照抗体INab308相比，全长抗C5a抗体RH23、RH25、RH32、RH35、RH36、RH58、RH85、RH86、RH92、RH95(重构建成人IgG4形式)与人C5a的结合能力更优。The results are shown in Figure 1A-1C. Compared with the control antibody INab308, the full-length anti-C5a antibodies RH23, RH25, RH32, RH35, RH36, RH58, RH85, RH86, RH92, RH95 (reconstituted human IgG4 form) and human C5a The combination ability is better.
C5a-desArg ELISA结合实验：C5a-desArg ELISA binding experiment:
采用ELISA结合实验评价全长抗C5a抗体RH23、RH25、RH32、RH35、RH36、RH58、RH85、RH86、RH92、RH95(重构建成人IgG4形式)与人C5a-desArg的亲和力，并以INab308作为对照。简言之，将天然人C5a-desArg抗原(Complement，A145)溶解在PBS溶液中，按照0.1μg/孔包被96孔板，4℃过夜。在加入抗体之前，用TBST溶液洗涤96孔板，用5％牛奶37℃封闭1-2小时，再用TBST溶液洗涤。首先将每个抗体样品稀释至10μg/mL，随后以1：3比例进行梯度稀释。将梯度稀释后的样品分别加入96孔板中，每孔100μL，37℃孵育1小时。随后用TBST溶液洗涤6次，加入抗人Fc-AP抗体(1:2500)，每孔100μL，37℃孵育1小时。TBST洗板机洗涤6次后，每孔加入pNPP 50μL，37℃孵育10min，用3M NaOH终止反应。读取410nm处的吸光值，并通过PRISM生成结合曲线。An ELISA binding experiment was used to evaluate the affinity of full-length anti-C5a antibodies RH23, RH25, RH32, RH35, RH36, RH58, RH85, RH86, RH92, and RH95 (reconstructed into human IgG4 form) with human C5a-desArg, and INab308 was used as a control. Briefly, natural human C5a-desArg antigen (Complement, A145) was dissolved in PBS solution, coated on a 96-well plate at 0.1 μg/well, and incubated at 4°C overnight. Before adding the antibody, wash the 96-well plate with TBST solution, block with 5% milk at 37°C for 1-2 hours, and then wash with TBST solution. Each antibody sample was first diluted to 10 μg/mL, followed by gradient dilution in a 1:3 ratio. Add the gradient diluted samples to a 96-well plate, 100 μL per well, and incubate at 37°C for 1 hour. Then wash with TBST solution 6 times, add anti-human Fc-AP antibody (1:2500), 100 μL per well, and incubate at 37°C for 1 hour. After washing with TBST plate washer 6 times, add 50 μL of pNPP to each well, incubate at 37°C for 10 min, and terminate the reaction with 3M NaOH. The absorbance value at 410 nm was read and a binding curve was generated by PRISM.
结果如图2所示，全长抗C5a抗体RH23、RH25、RH32、RH35、RH36、RH58、RH85、RH86、RH92、RH95(重构建成人IgG4形式)均能够很好的与C5a-desArg结合；且与对照抗体INab308相比，其与C5a-desArg的结合能力更优或与之相当。The results are shown in Figure 2. The full-length anti-C5a antibodies RH23, RH25, RH32, RH35, RH36, RH58, RH85, RH86, RH92, and RH95 (reconstructed into human IgG4 form) can all bind to C5a-desArg well; and Compared with the control antibody INab308, its binding ability to C5a-desArg is better or equivalent.
2.3活性氧(ROS)释放试验2.3 Reactive oxygen species (ROS) release test
C5a可刺激中性粒细胞释放活性氧(ROS)，从而促进中性粒细胞参与广泛的炎症反应。基于此机理，利用诱导型中性粒细胞检测抗C5a抗体的阻断作用。HL-60细胞系为人早幼粒细胞白血病细胞，简言之，使用1mM二丁酰cAMP钠盐(sigma,D0260)处理HL60细胞48h，以诱导其分化，细胞缩小呈梭形，并向中性粒细胞分化。C5a以剂量依赖的方式刺激分化的HL-60细胞产生ROS。C5a can stimulate neutrophils to release reactive oxygen species (ROS), thereby promoting neutrophils to participate in a wide range of inflammatory responses. Based on this mechanism, induced neutrophils were used to detect the blocking effect of anti-C5a antibodies. The HL-60 cell line is human promyelocytic leukemia cells. Briefly, HL60 cells were treated with 1mM dibutyryl cAMP sodium salt (sigma, D0260) for 48 hours to induce their differentiation. The cells shrank to a spindle shape and became neutral. Granulocytic differentiation. C5a stimulates ROS production in differentiated HL-60 cells in a dose-dependent manner.
分别将倍比稀释的抗C5a抗体RH23、RH25、RH32、RH35、RH36、RH58、RH85、RH86、RH92、RH95、对照抗体INab308(重构建成人IgG4形式)与C5a(5nM)预先混合，并用混合液处理分化的细胞30分钟。加入DCFH-DA荧光探针(sigma,D6883)，孵育。利用酶标仪检测在激发波长480nm，发射波长525nm条件下的荧光强度。将仅C5a刺激和无C5a刺激时的抑制率分别计为0％和100％，归一化实验数据，计算出C5a抗体的IC50值。Premix the diluted anti-C5a antibodies RH23, RH25, RH32, RH35, RH36, RH58, RH85, RH86, RH92, RH95, control antibody INab308 (reconstructed into adult IgG4 form) and C5a (5nM) respectively, and use the mixed solution Treat differentiated cells for 30 min. Add DCFH-DA fluorescent probe (sigma, D6883) and incubate. Use a microplate reader to detect the fluorescence intensity under the conditions of excitation wavelength 480nm and emission wavelength 525nm. The inhibition rates with only C5a stimulation and without C5a stimulation were calculated as 0% and 100%, respectively. The experimental data were normalized to calculate the IC 50 value of the C5a antibody.
结果如表5所示，抗C5a抗体RH23、RH25、RH32、RH35、RH36、RH58、RH85、RH86、RH92、RH95(重构建成人IgG4形式)均能够良好的抑制ROS释放，且与对照抗体INab308相当。The results are shown in Table 5. Anti-C5a antibodies RH23, RH25, RH32, RH35, RH36, RH58, RH85, RH86, RH92, and RH95 (reconstituted in the form of adult IgG4) can all inhibit ROS release well and are comparable to the control antibody INab308. .
表5 table 5
实施例3：抗C5a抗体治疗冠状病毒导致的ARDS的体内效果Example 3: In vivo effect of anti-C5a antibody in treating ARDS caused by coronavirus
构建ARDS动物疾病模型，评估抗C5a抗体在体内的治疗效果。ARDS动物疾病模型和正常对照组：共采用46只C5a人源化的小鼠(购自上海南方模式生物科技股份有限公司)，在实验前4天、3天和2天，向其中的40只小鼠注射携带并表达SARS-CoV-2 N蛋白的腺病毒(可参见Ting Gao等，https://doi.org/10.1101/2020.03.29.20041962)，7.5×108PFU/100μL/只/次/天，疾病模型组分为模型对照组、高剂量组(10mg/kg)、中剂量组(3mg/kg)和低剂量组(1mg/kg)。在第0天时(实验当天)向小鼠注射括号内相应剂量的抗体。剩余的6只小鼠则注射100μL 0.9％氯化钠溶液，作为正常对照组。向疾病模型对照组和各实验组小鼠注射LPS-K235(Sigma-Aldrich)，浓度1mg/mL，100μL/只。本研究中所有的试剂以尾静脉注射的方式施用。在给药后12h，24h，36h，48h，60h和72h时分别观察和分析各组小鼠的存活情况。抗体给药后72小时，将小鼠麻醉，眼眶采血。采用系列血液分析仪进行全血白细胞计数及分类，包括白细胞计数(WBC)、中性粒细胞(Neut)、淋巴细胞(Lymph)、单核细胞(Mono)。采用ELISA方法检测GM-CSF、IL-1β、IL-6、TNF-α、MCP-1等细胞因子水平。Construct an ARDS animal disease model and evaluate the therapeutic effect of anti-C5a antibodies in vivo. ARDS animal disease model and normal control group: A total of 46 C5a humanized mice (purchased from Shanghai Southern Model Biotechnology Co., Ltd.) were used. 40 of them were administered 4, 3 and 2 days before the experiment. Mice were injected with adenovirus carrying and expressing SARS-CoV-2 N protein (see Ting Gao et al., https://doi.org/10.1101/2020.03.29.20041962), 7.5×10 8 PFU/100 μL/mouse/time/ day, the disease model groups were divided into model control group, high-dose group (10mg/kg), medium-dose group (3mg/kg) and low-dose group (1mg/kg). On day 0 (the day of the experiment), mice were injected with the corresponding doses of antibodies in parentheses. The remaining 6 mice were injected with 100 μL of 0.9% sodium chloride solution as a normal control group. Mice in the disease model control group and each experimental group were injected with LPS-K235 (Sigma-Aldrich) at a concentration of 1 mg/mL, 100 μL/mouse. All reagents in this study were administered by tail vein injection. The survival of mice in each group was observed and analyzed at 12h, 24h, 36h, 48h, 60h and 72h after administration. 72 hours after antibody administration, the mice were anesthetized, and blood was collected from the orbits. use A series of blood analyzers perform whole blood white blood cell counting and classification, including white blood cell count (WBC), neutrophils (Neut), lymphocytes (Lymph), and monocytes (Mono). ELISA method was used to detect the levels of GM-CSF, IL-1β, IL-6, TNF-α, MCP-1 and other cytokines.
本申请的抗C5a抗体RH86能够有效降低或防止冠状病毒所引起的小鼠死亡，提高小鼠的存活率。与正常对照组相比，模型对照组中小鼠的WBC、Lymph及Mono的水平降低，GM-CSF、IL-1β、IL-6、TNF-α、MCP-1水平均显著升高，二者之间有统计学显著差异(P<0.05)。与疾病模型对照组比较，施用抗C5a抗体(RH86)的三个剂量实验组中均显示出WBC和Lymph数量的升高，GM-CSF、IL-1β、IL-6、TNF-α、MCP-1、C5a水平均呈剂量依赖性的下降。上述结果表明，本申请的抗C5a抗体RH86有助于恢复ARDS疾病模型小鼠体内免疫细胞的平衡状态，能够在体内显著降低新型冠状病毒导致的细胞因子风暴和炎症反应(数据未展示)。The anti-C5a antibody RH86 of the present application can effectively reduce or prevent the death of mice caused by coronavirus and improve the survival rate of mice. Compared with the normal control group, the levels of WBC, Lymph and Mono of mice in the model control group were reduced, and the levels of GM-CSF, IL-1β, IL-6, TNF-α, and MCP-1 were significantly increased. There is a statistically significant difference between them (P<0.05). Compared with the disease model control group, the three dose experimental groups administered anti-C5a antibody (RH86) all showed an increase in the number of WBC and Lymph, GM-CSF, IL-1β, IL-6, TNF-α, MCP- 1. C5a levels decreased in a dose-dependent manner. The above results show that the anti-C5a antibody RH86 of the present application can help restore the balance of immune cells in ARDS disease model mice and can significantly reduce the cytokine storm and inflammatory response caused by the new coronavirus in vivo (data not shown).

Claims (23)

  1. 一种分离的特异性识别C5a的抗体，其中所述抗C5a抗体包含：An isolated antibody that specifically recognizes C5a, wherein the anti-C5a antibody comprises:
    (i)VH，其包含如氨基酸序列SEQ ID NO:25所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；(i) VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO: 25;
    以及VL，其包含如氨基酸序列SEQ ID NO:35所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；and VL , which comprises LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO:35;
    (ii)VH，其包含如氨基酸序列SEQ ID NO:26所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；(ii) VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO: 26;
    以及VL，其包含如氨基酸序列SEQ ID NO:36所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；and VL , which comprises LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO:36;
    (iii)VH，其包含如氨基酸序列SEQ ID NO:27所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；(iii) VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO: 27;
    以及VL，其包含如氨基酸序列SEQ ID NO:37所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；and VL , which comprises LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO:37;
    (iv)VH，其包含如氨基酸序列SEQ ID NO:28所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；(iv) VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO: 28;
    以及VL，其包含如氨基酸序列SEQ ID NO:38所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；and VL , which comprises LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO:38;
    (v)VH，其包含如氨基酸序列SEQ ID NO:29所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；(v) VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO: 29;
    以及VL，其包含如氨基酸序列SEQ ID NO:39所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；and VL , which comprises LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO:39;
    (vi)VH，其包含如氨基酸序列SEQ ID NO:30所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；(vi) VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO: 30;
    以及VL，其包含如氨基酸序列SEQ ID NO:40所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；and VL , which comprises LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO:40;
    (vii)VH，其包含如氨基酸序列SEQ ID NO:31所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；(vii) V H comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by V H as shown in the amino acid sequence SEQ ID NO: 31;
    以及VL，其包含如氨基酸序列SEQ ID NO:41所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3； and VL , which comprises LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO: 41;
    (viii)VH，其包含如氨基酸序列SEQ ID NO:32所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；(viii) V H comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by V H as shown in the amino acid sequence SEQ ID NO: 32;
    以及VL，其包含如氨基酸序列SEQ ID NO:42所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；and VL , which comprises LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO:42;
    (ix)VH，其包含如氨基酸序列SEQ ID NO:33所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；(ix) VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO: 33;
    以及VL，其包含如氨基酸序列SEQ ID NO:43所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3；或and a VL comprising LC-CDR1, LC-CDR2 and LC-CDR3 comprised by VL as shown in the amino acid sequence SEQ ID NO: 43; or
    (x)VH，其包含如氨基酸序列SEQ ID NO:34所示的VH包含的HC-CDR1、HC-CDR2和HC-CDR3；以及VL，其包含如氨基酸序列SEQ ID NO:44所示的VL包含的LC-CDR1、LC-CDR2和LC-CDR3。(x) VH comprising HC-CDR1, HC-CDR2 and HC-CDR3 comprised by VH as shown in the amino acid sequence SEQ ID NO:34; and VL comprising as shown in the amino acid sequence SEQ ID NO:44 The V L shown contains LC-CDR1, LC-CDR2 and LC-CDR3.
  2. 一种分离的特异性识别C5a的抗体，其中所述抗体包含：An isolated antibody that specifically recognizes C5a, wherein the antibody comprises:
    (i)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:2，和HC-CDR3，其包含氨基酸序列SEQ ID NO:6，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(i) V H comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 2, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 6, or a variant of the V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO: 12. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 20, or a variant of the V L , which contains up to about 5 LC-CDRs. substitution of amino acids;
    (ii)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:20，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(ii) V H comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO: 12. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 20, or a variant of the V L , which contains up to about 5 LC-CDRs. substitution of amino acids;
    (iii)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:8，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:13，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:21，或 者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(iii) V H comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 8, or a variant of the V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 13. LC-CDR2, which contains the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which contains the amino acid sequence SEQ ID NO: 21, or A variant of the V L whose LC-CDRs contain at most about 5 amino acid substitutions;
    (iv)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(iv) V H comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 9, or a variant of the V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 14. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or a variant of the V L , which contains up to about 5 LC-CDRs. substitution of amino acids;
    (v)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:5，和HC-CDR3，其包含氨基酸序列SEQ ID NO:10，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(v) V H comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 5, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 10, or a variant of the V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 14. LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , which contains up to about 5 LC-CDRs substitution of amino acids;
    (vi)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(vi) V H comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO: 14. LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , which contains up to about 5 LC-CDRs substitution of amino acids;
    (vii)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:9，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:12，LC-CDR2，其包含氨基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:23，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(vii) V H comprising: HC-CDR1 comprising the amino acid sequence SEQ ID NO: 1, HC-CDR2 comprising the amino acid sequence SEQ ID NO: 4, and HC-CDR3 comprising the amino acid sequence SEQ ID NO: 9, or a variant of the V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 12. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 23, or a variant of the V L , which contains up to about 5 LC-CDRs. substitution of amino acids;
    (viii)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:15，LC-CDR2，其包含氨 基酸序列SEQ ID NO:17，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；(viii) V H comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO: 15. LC-CDR2, which contains ammonia The amino acid sequence SEQ ID NO: 17, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 22, or a variant of the V L whose LC-CDRs contain up to about 5 amino acid substitutions;
    (ix)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:4，和HC-CDR3，其包含氨基酸序列SEQ ID NO:7，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:14，LC-CDR2，其包含氨基酸序列SEQ ID NO:18，和LC-CDR3，其包含氨基酸序列SEQ ID NO:22，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代；或(ix) V H comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 4, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 7, or a variant of the V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC-CDR1 comprising the amino acid sequence SEQ ID NO: 14. LC-CDR2, which comprises the amino acid sequence SEQ ID NO: 18, and LC-CDR3, which comprises the amino acid sequence SEQ ID NO: 22, or a variant of said V L , which contains up to about 5 LC-CDRs Substitution of amino acids; or
    (x)VH，所述VH包含：HC-CDR1，其包含氨基酸序列SEQ ID NO:1，HC-CDR2，其包含氨基酸序列SEQ ID NO:3，和HC-CDR3，其包含氨基酸序列SEQ ID NO:11，或者所述VH的变体，其HC-CDRs中包含至多约5个氨基酸的取代；以及VL，所述VL包含：LC-CDR1，其包含氨基酸序列SEQ ID NO:16，LC-CDR2，其包含氨基酸序列SEQ ID NO:19，和LC-CDR3，其包含氨基酸序列SEQ ID NO:24，或者所述VL的变体，其LC-CDRs中包含至多约5个氨基酸的取代。(x) V H comprising: HC-CDR1, which comprises the amino acid sequence SEQ ID NO: 1, HC-CDR2, which comprises the amino acid sequence SEQ ID NO: 3, and HC-CDR3, which comprises the amino acid sequence SEQ ID NO: 11, or a variant of the V H comprising up to about 5 amino acid substitutions in its HC-CDRs; and V L comprising: LC- CDR1 comprising the amino acid sequence SEQ ID NO: 16. LC-CDR2, which includes the amino acid sequence SEQ ID NO: 19, and LC-CDR3, which includes the amino acid sequence SEQ ID NO: 24, or a variant of the V L , which contains up to about 5 LC-CDRs. Amino acid substitutions.
  3. 根据权利要求1或2中所述的分离的特异性识别C5a的抗体，其包含：The isolated antibody specifically recognizing C5a according to claim 1 or 2, comprising:
    (i)VH，其包含SEQ ID NO:25所示的氨基酸序列或其变体，所述变体与SEQ ID NO:25所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:35所示的氨基酸序列或其变体，所述变体与SEQ ID NO:35所示的氨基酸序列具有至少约80％序列同一性；(i) V H comprising the amino acid sequence set forth in SEQ ID NO: 25 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 25; and V L , which comprises the amino acid sequence shown in SEQ ID NO:35 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:35;
    (ii)VH，其包含SEQ ID NO:26所示的氨基酸序列或其变体，所述变体与SEQ ID NO:26所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:36所示的氨基酸序列或其变体，所述变体与SEQ ID NO:36所示的氨基酸序列具有至少约80％序列同一性；(ii) V H comprising the amino acid sequence set forth in SEQ ID NO: 26 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 26; and V L , which comprises the amino acid sequence shown in SEQ ID NO:36 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:36;
    (iii)VH，其包含SEQ ID NO:27所示的氨基酸序列或其变体，所述变体与SEQ ID NO:27所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:37所示的氨基酸序列或其变体，所述变体与SEQ ID NO:37所示的氨基酸序列具有至少约80％序列同一性；(iii) V H comprising the amino acid sequence set forth in SEQ ID NO: 27 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 27; and V L , which comprises the amino acid sequence shown in SEQ ID NO: 37 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO: 37;
    (iv)VH，其包含SEQ ID NO:28所示的氨基酸序列或其变体，所述变体与SEQ ID NO:28所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ  ID NO:38所示的氨基酸序列或其变体，所述变体与SEQ ID NO:38所示的氨基酸序列具有至少约80％序列同一性；(iv) V H comprising the amino acid sequence set forth in SEQ ID NO: 28 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 28; and V L , which contains SEQ The amino acid sequence shown in ID NO:38 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:38;
    (v)VH，其包含SEQ ID NO:29所示的氨基酸序列或其变体，所述变体与SEQ ID NO:29所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:39所示的氨基酸序列或其变体，所述变体与SEQ ID NO:39所示的氨基酸序列具有至少约80％序列同一性；(v) V H comprising the amino acid sequence set forth in SEQ ID NO: 29 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 29; and V L , which comprises the amino acid sequence shown in SEQ ID NO:39 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:39;
    (vi)VH，其包含SEQ ID NO:30所示的氨基酸序列或其变体，所述变体与SEQ ID NO:30所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:40所示的氨基酸序列或其变体，所述变体与SEQ ID NO:40所示的氨基酸序列具有至少约80％序列同一性；(vi) V H comprising the amino acid sequence set forth in SEQ ID NO: 30 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 30; and V L , which comprises the amino acid sequence shown in SEQ ID NO:40 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:40;
    (vii)VH，其包含SEQ ID NO:31所示的氨基酸序列或其变体，所述变体与SEQ ID NO:31所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:41所示的氨基酸序列或其变体，所述变体与SEQ ID NO:41所示的氨基酸序列具有至少约80％序列同一性；(vii) V H comprising the amino acid sequence set forth in SEQ ID NO: 31 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 31; and V L , which comprises the amino acid sequence shown in SEQ ID NO:41 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:41;
    (viii)VH，其包含SEQ ID NO:32所示的氨基酸序列或其变体，所述变体与SEQ ID NO:32所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:42所示的氨基酸序列或其变体，所述变体与SEQ ID NO:42所示的氨基酸序列具有至少约80％序列同一性；(viii) V H comprising the amino acid sequence set forth in SEQ ID NO: 32 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 32; and V L , which comprises the amino acid sequence shown in SEQ ID NO:42 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:42;
    (ix)VH，其包含SEQ ID NO:33所示的氨基酸序列或其变体，所述变体与SEQ ID NO:33所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:43所示的氨基酸序列或其变体，所述变体与SEQ ID NO:43所示的氨基酸序列具有至少约80％序列同一性；或(ix) V H comprising the amino acid sequence set forth in SEQ ID NO: 33 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 33; and V L , which comprises the amino acid sequence shown in SEQ ID NO:43 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:43; or
    (x)VH，其包含SEQ ID NO:34所示的氨基酸序列或其变体，所述变体与SEQ ID NO:34所示的氨基酸序列具有至少约80％序列同一性；以及VL，其包含SEQ ID NO:44所示的氨基酸序列或其变体，所述变体与SEQ ID NO:44所示的氨基酸序列具有至少约80％序列同一性。(x) V H comprising the amino acid sequence set forth in SEQ ID NO: 34 or a variant thereof that has at least about 80% sequence identity with the amino acid sequence set forth in SEQ ID NO: 34; and V L , which comprises the amino acid sequence shown in SEQ ID NO:44 or a variant thereof, which variant has at least about 80% sequence identity with the amino acid sequence shown in SEQ ID NO:44.
  4. 根据权利要求1-3中任一项的分离的特异性识别C5a的抗体，其中所述抗体包含Fc片段。An isolated antibody specifically recognizing C5a according to any one of claims 1-3, wherein said antibody comprises an Fc fragment.
  5. 根据权利要求4中的分离的特异性识别C5a的抗体，其中所述抗体是全长的IgG 抗体。An isolated antibody specifically recognizing C5a according to claim 4, wherein said antibody is a full-length IgG Antibody.
  6. 根据权利要求5中的分离的特异性识别C5a的抗体，其中所述抗体是全长的IgG1、IgG2、IgG3或IgG4抗体。An isolated antibody specifically recognizing C5a according to claim 5, wherein said antibody is a full-length IgGl, IgG2, IgG3 or IgG4 antibody.
  7. 根据权利要求1-6中任一项的分离的特异性识别C5a的抗体，其中所述抗体是嵌合的、人源化的或全人的抗体。An isolated antibody specifically recognizing C5a according to any one of claims 1 to 6, wherein said antibody is a chimeric, humanized or fully human antibody.
  8. 根据权利要求1-3中任一项的分离的特异性识别C5a的抗体是抗原结合片段，其中所述抗原结合片段选自Fab、Fab’、F(ab)’2、Fab’-SH、单链抗体(scFv)、Fv片段、dAb、Fd、纳米抗体(nanobody)、双链抗体(diabody)和线性抗体。The isolated antibody that specifically recognizes C5a according to any one of claims 1-3 is an antigen-binding fragment, wherein the antigen-binding fragment is selected from the group consisting of Fab, Fab', F(ab)' 2 , Fab'-SH, single Chain antibodies (scFv), Fv fragments, dAb, Fd, nanobodies, diabodies and linear antibodies.
  9. 一种核酸分子，其编码权利要求1-8中任一项所述的特异性识别C5a的抗体。A nucleic acid molecule encoding the antibody specifically recognizing C5a according to any one of claims 1-8.
  10. 一种包含权利要求9中所述的核酸分子的载体。A vector comprising the nucleic acid molecule of claim 9.
  11. 一种分离的宿主细胞，其包含权利要求1-8中任一项所述的特异性识别C5a的抗体、权利要求9中所述的核酸分子或权利要求10中所述的载体。An isolated host cell comprising the antibody specifically recognizing C5a according to any one of claims 1-8, the nucleic acid molecule according to claim 9 or the vector according to claim 10.
  12. 一种制备特异性识别C5a的抗体的方法，其包含：A method of preparing an antibody that specifically recognizes C5a, comprising:
    a)在能有效表达特异性识别C5a的抗体的条件下培养权利要求11中所述的宿主细胞；和a) culturing the host cell described in claim 11 under conditions that can effectively express an antibody that specifically recognizes C5a; and
    b)从宿主细胞中获得所表达的特异性识别C5a的抗体。b) Obtaining the expressed antibody that specifically recognizes C5a from the host cell.
  13. 一种药物组合物，其包含权利要求1-8中任一项所述的特异性识别C5a的抗体、权利要求9中所述的核酸分子、权利要求10中所述的载体或权利要求11中所述的分离的宿主细胞，以及药学上可接受的载体。A pharmaceutical composition comprising the antibody specifically recognizing C5a described in any one of claims 1-8, the nucleic acid molecule described in claim 9, the vector described in claim 10 or the vector described in claim 11 The isolated host cells, and pharmaceutically acceptable carriers.
  14. 权利要求1-8中任一项所述的抗体、权利要求9中所述的核酸分子、权利要求10中所述的载体、权利要求11中所述的分离的宿主细胞、由权利要求12中所述方法制备得到的抗体、或权利要求13中所述的药物组合物在制备治疗、预防和/或改善疾病或病症的药物中的用途。The antibody of any one of claims 1 to 8, the nucleic acid molecule of claim 9, the vector of claim 10, the isolated host cell of claim 11, the nucleic acid molecule of claim 12 The use of the antibody prepared by the method or the pharmaceutical composition described in claim 13 in the preparation of drugs for treating, preventing and/or ameliorating diseases or conditions.
  15. 根据权利要求14中的用途，其中所述疾病或病症为自身免疫性疾病、炎症、癌症、疼痛、呼吸和/或移植相关疾病。14. Use according to claim 14, wherein the disease or condition is an autoimmune disease, inflammation, cancer, pain, respiratory and/or transplant-related disease.
  16. 根据权利要求15中的用途，其中所述疾病或病症选自炎症反应综合征(SIRS)、败血症、严重败血症、感染性休克、缺血/再灌注相关损伤、急性肺损伤、肺炎、移植患者中急性和慢性移植排斥、移植物抗宿主反应、肾小球疾病、肾小球肾炎、实体肾功能衰竭、风湿性关节炎、自身免疫性疾病、Bechterew氏病、狼疮类疾病、炎症性肠病、克罗恩氏病、 肿瘤生长和实体器官癌症。Use according to claim 15, wherein the disease or condition is selected from the group consisting of inflammatory response syndrome (SIRS), sepsis, severe sepsis, septic shock, ischemia/reperfusion related injury, acute lung injury, pneumonia, transplant patients Acute and chronic transplant rejection, graft-versus-host reaction, glomerular disease, glomerulonephritis, solid renal failure, rheumatoid arthritis, autoimmune diseases, Bechterew's disease, lupus-like diseases, inflammatory bowel disease, Crohn's disease, Tumor growth and solid organ cancers.
  17. 一种偶联物，其包含单克隆抗体以及偶联部分，其中所述单克隆抗体为权利要求1-8中任一项所述的特异性识别C5a的抗体，所述偶联部分为放射性同位素、荧光物质、发光物质、有色物质或酶。A conjugate comprising a monoclonal antibody and a coupling portion, wherein the monoclonal antibody is the antibody specifically recognizing C5a according to any one of claims 1-8, and the coupling portion is a radioactive isotope , fluorescent substances, luminescent substances, colored substances or enzymes.
  18. 一种检测样品中C5a蛋白存在或其水平的制品或试剂盒，其包含权利要求1-8中任一项所述的特异性识别C5a的抗体、或权利要求17中所述的偶联物。A product or kit for detecting the presence or level of C5a protein in a sample, which contains the antibody specifically recognizing C5a according to any one of claims 1 to 8, or the conjugate described in claim 17.
  19. 权利要求1-8中任一项所述的特异性识别C5a的抗体、或权利要求17中所述的偶联物在制备用于检测C5a存在或水平的制品或试剂盒中的用途。Use of the antibody specifically recognizing C5a according to any one of claims 1 to 8, or the conjugate described in claim 17 in the preparation of products or kits for detecting the presence or level of C5a.
  20. 一种检测样品中C5a存在或水平的体外方法，所述方法包括：a)在允许抗体和C5a之间形成复合物的条件下将样品与权利要求1-8中任一项所述的特异性识别C5a的抗体、或权利要求17中所述的偶联物相接触；b)检测复合物的形成；和c)确定样品中C5a的存在或其水平。An in vitro method for detecting the presence or level of C5a in a sample, the method comprising: a) contacting the sample with the specificity of any one of claims 1-8 under conditions that allow the formation of a complex between the antibody and C5a contacting an antibody that recognizes C5a, or a conjugate as claimed in claim 17; b) detecting the formation of a complex; and c) determining the presence or level of C5a in the sample.
  21. 一种诊断C5a水平异常的方法，所述方法包括：a)获得待测样品，b)在允许抗体和C5a之间形成复合物的条件下将待测样品与权利要求1-8中任一项所述的特异性识别C5a的抗体、或权利要求17中所述的偶联物相接触；c)检测复合物的形成；d)确定待测样品中C5a的水平；和e)判断待测样品中C5a的水平是否高于正常组样品中C5a的水平。A method for diagnosing abnormal C5a levels, the method comprising: a) obtaining a sample to be tested, b) combining the sample to be tested with any one of claims 1-8 under conditions that allow the formation of a complex between the antibody and C5a The antibody specifically recognizing C5a, or the conjugate described in claim 17 is contacted; c) detecting the formation of the complex; d) determining the level of C5a in the sample to be tested; and e) judging the sample to be tested Whether the level of C5a in the sample is higher than the level of C5a in the normal group sample.
  22. 根据权利要求21中所述的方法，其中所述待测样品来源于有需要的患者，进一步地，所述患者已接受过或未接受过C5a抗体治疗。The method according to claim 21, wherein the sample to be tested is derived from a patient in need, and further, the patient has received or has not received C5a antibody treatment.
  23. 根据权利要求18中所述的制品或试剂盒、权利要求20中所述的检测方法或权利要求21-22中任一项所述的诊断方法，其中所述样品为血清样品或血浆样品。 The article or kit of claim 18, the detection method of claim 20 or the diagnostic method of any one of claims 21-22, wherein the sample is a serum sample or plasma sample.
PCT/CN2023/085301 2022-04-02 2023-03-31 Antibody specifically recognizing c5a and application of antibody WO2023186054A1 (en)

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