Antibodies Targeting C5ar

Abstract

The present invention provides novel antibodies or antibody fragments specifically binding to human C5aR. In particular, it relates to antibodies or antibody fragments that have combined beneficial properties and are therefore useful for the treatment of inflammatory or autoimmune diseases or cancer.

Description

FIELD OF THE INVENTION

[0001] The present disclosure relates to antibodies, which interact with C5aR, in particular with human C5aR. The present disclosure also relates to nucleic acid compositions, vector composition and host cells capable of expressing said antibodies, pharmaceutical compositions comprising said antibodies and uses of said antibodies for the treatment of specific diseases and/or for diagnostic purposes.

BACKGROUND

[0002] The C5a anaphylatoxin chemotactic receptor 1 (C5aR) (also known as CD88) is a G-protein-coupled receptor (GPCR) belonging to the rhodopsin family and is one of the two high-affinity receptors for the ligand, C5a, which is produced in serum as one of the core effector components of the complement response. Under physiological conditions, C5a acts as chemotactic agent for inflammatory cells, stimulates their respiratory burst as well as cytokine and chemokine release, and functions to increase vascular permeability.

[0003] C5aR appears widely expressed by various cell types. Highest C5aR expression levels are described for neutrophils. Low-to-moderate expression levels have been shown for macrophages/monocytes, dendritic cells, mast cells, eosinophils, lung vascular smooth muscle cells, astrocytes, microglia, osteoblasts, osteoclasts, epithelial and endothelial cells (Monk P N et al., Br J Pharmacol. 2007, 152: 429-448; Wetsel R A, Immunol Lett. 1995, 44: 183-187). For human T cells, low expression levels have been observed (Nataf S, J Immunol. 1999, 162: 4018-4023).

[0004] Cell responses to C5a are tightly controlled by ligand-induced receptor internalization. C5aR is described to rapidly and dose-dependently internalize upon C5a treatment and up to 90% of the receptor is recycled back to the cell surface. Thus, the high expression levels of C5aR in combination with its fast turn-over rate could be limiting in terms of efficacy due to target-mediated drug disposition (TMDD) effects.

[0005] The interaction of C5aR with C5a has been described in many different disease settings; most of them being involved in inflammatory and autoimmune diseases (Morgan B P et al., Nat Rev Drug Discov. 2015, 14: 857-877; Hawksworth O A et al., Mol Immunol. 2017, 89: 36-43). Some initial research has been performed to identify the underlying mechanisms by which C5a stimulates tumor growth (Markiewski M M et al., Nat Immunol. 2008, 9: 1225-1235; Corrales L. et al, J Immunol. 2012, 189: 4674-4683; Cho M S et al., Cell Rep. 2014, 6: 1085-1095). Most of the data implicates that C5a increases cancer cell proliferation, intra-tumor angiogenesis and enhances tumor invasiveness and metastasis. More recent data also implicate a role for C5a/C5aR in the generation of immunosuppressive environments in the context of solid tumors (Sayegh E T et al., Cancer Med. 2014, 3: 747-758; Darling V R et al., Expert Rev Clin Immunol. 2015, 11: 255-263; Markiewski M M et al., Cancer Res. 2009, 69: 6367-6370) resulting in enhanced primary tumor growth by inhibiting antitumor responses (e.g. increased recruitment of C5aR-expressing myeloid cells such as myeloid-derived suppressor cell (MDSC) or M2 macrophages). Based on these findings, combination strategies with already known anti-tumor agents, such as immune checkpoint protein inhibitors in order to boost a subject's immune response by reducing the immunosuppressive microenvironment are in focus of research and clinical development (Wang Y, et al.: Cancer Discov. 2016, 6: 1022-1035).

[0006] To date, only one specific complement therapeutic has been approved, which targets the upstream molecule of C5a, namely C5. The humanized anti-C5 mAb Eculizumab is capable of binding C5, preventing its cleavage and formation of C5a and C5b proteins, and subsequent MAC formation. Various other therapeutic monoclonal C5 specific antibodies, such as ALXN1210 or LFG316 are under clinical evaluation (Hawksworth O A et al., Mol Immunol. 2017, 89: 36-43). However, since increased infection risk is a major concern with chronic C5 treatment, specific targeting of downstream molecules whilst preventing the biological activities of other complement components is clearly advantageous. Accordingly, a number of antagonistic C5a specific monoclonal antibodies are under development.

[0007] Direct targeting of C5aR has a number of advantages over targeting C5 or C5a, respectively. First, the inhibition of the receptor alone would preserve MAC activity, thereby reducing the potential risk of infections. Second, C5aR blockade permits continued C5a interaction with its second receptor C5L2. Since C5L2 has been reported to have anti-inflammatory effects, maintaining an effective C5L2 signalling pathway may result in increased efficacy or reduced dosing requirements. Third, direct C5aR targeting may provide pharmacodynamic advantages over inhibition of soluble C5a, due to its small molecular weight and its high turnover rate. Overall, there is strong interest in developing C5aR inhibitors, such as aptamers, peptides, and non-peptide small molecules being tested in pre-clinical and clinical trials.

[0008] Neutralizing polyclonal antiserum or monoclonal antibodies directed against the N-terminal extracellular region of human C5aR and being able to interfere with C5aR-C5a interaction has been described in the art (see e.g. Morgan et al., The Journal of Immunology, Vol 151, 377-388. No. 1, Jul. 1, 1993; Oppermann et al., The Journal of Immunology, Vol 151, 3785-3794, No. 7, Oct. 1, 1993),

[0009] However, these C5aR specific antibodies are not suited for the clinical development and therapeutic use in human, especially due to their animal origin (which makes them immunogenic in human patients), clonality, and/or lack of cross-reactivity to relevant animal species.

[0010] Therapeutic antibodies targeting C5aR has been considered for clinical development. However, the clinical development of the antagonistic C5aR specific antibody Neutrazumab, a humanized IgG4 mAb was stopped in Phase II clinical trials due to issues with immune cell depletion and immunogenicity (Daniluk S et al., Annals of the Rheumatic Diseases. 2014, 73: 684-685). Since C5aR appears constitutively expressed on a variety of cell types, it is important that an antagonistic antibody does not induce any depletion of the target cells.

[0011] To overcome the limitations of Neutrazumab, a second generation of a C5aR specific antibody has been generated, namely NNC0215-0384 (US2013/0295116 (NOVO NORDISK); clone 32F3A6GL). This antibody is a human IgG1 antibody derived from transgenic mice and is currently under clinical development as IPH5401 in the field of cancer (Olivier Demaria et al., Innate Pharma 2017. Poster #B184. CRI-CIMT-EATI-AACR Mainz). IPH5401 bears a silenced human IgG1 Fc region to eliminate the ability of the antibody to induce effector function. This antibody is herein referred to as RefMAB#1.

SUMMARY OF THE INVENTION

[0012] The present disclosure provides novel antibodies and antibody fragments.

[0013] The antibodies and antibody fragments disclosed herein can specifically bind to human C5aR and preferably cross-react with C5aR from cynomolgus monkey. Accordingly, in some embodiments, the disclosed antibodies are specific for human C5aR and cynomolgus C5aR. In some other embodiments, the disclosed antibodies or antibody fragments bind to the N-terminal extracellular region of human and cynomolgus C5aR.

[0014] This is in contrast to the above referenced prior art antibody IPH5401, which binds to the second extracellular loop of human C5aR resulting in the lack of binding to cynomolgus monkey C5aR, a commonly used relevant toxicology species.

[0015] In addition, the inventors of the present invention surprisingly found that the presently claimed C5aR specific antibodies are not only significantly more potent in neutralizing pathophysiological C5a concentrations when compared to IPH5401 but also revealed an increased potency over time in inhibiting C5 mediated activation of neutrophils in vitro.

[0016] Accordingly, in some embodiments, the disclosed antibodies can efficiently inhibit C5a induced C5aR activity in vitro, most notably at pathophysiological C5a concentration. The disclosed antibodies or antibody fragments may also inhibit C5a induced leucocyte activation in vitro as determined by their ability to inhibit C5a induced upregulation of CD11b in granulocytes and/or monocytes. In some embodiments, the disclosed antibodies or antibody fragments inhibit human C5a induced CD11b expression in human granulocytes with an IC.sub.50 concentration of 42 nM in the presence of 150 nM human C5a in vitro. In some other embodiments, the disclosed antibodies may exhibit an increased potency to inhibit C5a induced upregulation of CD11b in granulocytes and/or monocytes after a prolonged period of incubation time. The disclosed antibodies may be also efficient in inhibiting C5a induced neutrophil migration.

[0017] In sum, the present disclosure provides novel antibodies, which are superior to the C5aR specific antibodies known from the art. In particular, the antibodies of the present disclosure are human antibodies with high affinity binding to human C5aR, which preferably cross-react with cynomolgus monkey C5aR and have favourable functional and safety properties never have been observed before. These features makes the antibodies of the present disclosure highly desirable for therapeutic use such as for preventing and/or treating inflammatory and autoimmune diseases as well as cancer.

[0018] The present disclosure provides isolated antibodies or antibody fragments that specifically bind to human C5aR having CDR regions according to Table 1 or Table 2 of the present specification. The present disclosure also provides isolated antibodies or antibody fragments specific for human C5aR having a variable heavy chain region (VH) and a variable light chain region (VL) comprising the amino acid sequences according to Table 1 or Table 2 of the present specification. The present disclosure also provides isolated antibodies or antibody fragments specific for C5aR having a heavy chain (HC) and a light chain (LC) comprising the amino acid sequences according to Table 1 or Table 2 of the present specification.

[0019] The isolated antibodies of the present disclosure do not substantially induce effector function in vitro. Such effector function may comprise ADCP, ADCC or CDC. Furthermore, the isolated antibody or antibody fragments of the present disclosure comprise one or more amino acid substitution selected from the group of: L234A, L235E, G237A, A330S and P331S, with numbering according to EU index. In particular, the isolated antibodies or antibody fragments of the present disclosure comprise a variant human IgG1 Fc region, which comprises the following amino acid substitutions: L234A, L235E, G237A, A330S and P331S with numbering according EU index.

[0020] The present disclosure also provides the isolated antibodies or antibody fragments of the present disclosure for use in medicine.

[0021] The present disclosure also provides methods for treating a subject suffering from a disease, such as an inflammatory or autoimmune disease or cancer, by administering to said subject an effective amount of the antibodies or antibody fragments of the present disclosure. Preferably, said subject is a human.

[0022] The present disclosure also provides pharmaceutical compositions comprising the isolated antibodies or antibody fragments of the present disclosure, and a pharmaceutically acceptable carrier.

[0023] The present disclosure also provides nucleic acid compositions encoding the isolated antibodies or antibody fragments of the present disclosure. The present disclosure also provides vector compositions comprising the nucleic acid compositions encoding the isolated antibodies or antibody fragments of the present disclosure. The present disclosure also provides host cells comprising the vector compositions or nucleic acids compositions encoding the isolated antibodies or antibody fragments of the present disclosure.

[0024] The present disclosure also provides methods for treating a subject suffering from a disease, such as an inflammatory disease, autoimmune disease or cancer by administering to said subject an effective amount of the isolated antibodies or antibody fragments of the present disclosure. Preferably, said subject is a human.

[0025] The present disclosure also provides pharmaceutical compositions comprising the isolated antibodies or antibody fragments of the present disclosure, and a pharmaceutically acceptable carrier.

[0026] There is utility in the claimed antibodies or antibody fragments. Furthermore, there is utility in the claimed method to identify such antibodies or antibody fragments.

[0027] Utilization of the claimed antibodies or antibody fragments is to alter the biological activity of human C5aR. In particular, the claimed antibodies or antibody fragments are for therapeutic use, such as the treatment of inflammatory or autoimmune disease or cancer

BRIEF DESCRIPTION OF THE DRAWINGS

[0028] FIG. 1: Cell binding of MAB#1, MAB#2, RefMAB#1 and negative isotype control MOR03207 to human and cynomolgus monkey C5aR determined via FACS. A Dose response binding to human C5aR overexpressed on Flp-In CHO cells. B Dose response binding to cynomolgus monkey C5aR overexpressed on Flp-In CHO cells. C Average dose response binding to purified human neutrophils obtained from whole blood of three different donors. D Average dose response binding to purified cynomolgus monkey neutrophils obtained from whole blood from three different monkeys.

[0029] FIG. 2: Cell binding of MAB#1, MAB#2, RefMAB#1 and negative isotype control MOR03207 to human, cynomolgus and rodent C5aR as well as to the C5aR related GPCRs human C5L2, human C3aR, human FPR1 and human ChemR23 determined via FACS at an IgG concentration of 600 nM.

[0030] FIG. 3: ELISA binding of MAB#1 to two natural variants of human C5aR as well as to mouse C5aR expressed on virus-like-particles (VLPs).

[0031] FIG. 4: PathHunter.RTM.--.beta.-arrestin assay from DiscoveRx. Neutralization of human C5a induced .beta.-arrestin recruitment. A Log dose-response curves for increasing concentrations of recombinant human C5a in absence or presence of MAB#1 (50 nM), MAB#2 (50 nM) and RefMAB#1 (50 nM). B Percentage inhibition was calculated for three increasing concentrations of human C5a (1.2 nM, 11 nM and 100 nM, respectively) at a final IgG concentration of 50 nM.

[0032] FIG. 5: Inhibition of human C5a induced CD11b upregulation in human granulocytes at regular and pathophysiological C5a concentrations. A+B Comparison of MAB#1, MAB#2 and RefMAB#1 in a CD11b whole blood assay. Log dose-inhibition curves are shown. Human granulocytes were gated as target cells. As quantitative read-out, the IgG concentration needed to reach 50% inhibition of CD11b upregulation was calculated (IC.sub.50 concentration). IC.sub.50 concentrations for each IgG are depicted below the x-axis. A Log dose-response curves for increasing concentrations of IgG and 15 nM human C5a. B Log dose-response curves for increasing concentrations of IgG and 150 nM human C5a.

[0033] FIG. 6: Inhibition of human C5a induced CD11b upregulation in human granulocytes and monocytes determined over a prolonged period of time of incubation. Comparison of MAB#1 and RefMAB#1 in a CD11b whole blood assay after incubation of IgGs with target cells for either 20 minutes or 300 minutes. IgGs were added in serial dilutions and incubated for either 20 minutes or 300 minutes with subsequent stimulation with 15 nM human C5a. Either granulocytes (FIGS. 6A and B) or monocytes (FIGS. 6C and D) were gated as target cells. CD11b levels were determined. Log dose-inhibition curves are shown. Data are expressed as % inhibition of CD11b upregulation at 15 nM human C5a. Results for MAB#1 are shown in FIGS. 6A and C and results for RefMAB#1 are provided in FIGS. 6B and D.

[0034] FIG. 7: Inhibition of human C5a induced human neutrophil migration. MAB#1 and negative control MOR03207 were each tested at two IgG concentration (100 nM and 600 nM, respectively) in the presence of 10 nM human C5a. Average values from three independent assay runs at 3 different time points (15 min., 25 min., 35 min.) are shown. Neutrophils were obtained from 3 different human donors. Percentage inhibition was calculated on the basis of neutrophil migration in the absence of antibody.

[0035] FIG. 8: Promega ADCC and ADCP reporter bioassay. Comparison of MAB#1 and a monoclonal anti-C5aR control IgG bearing either a wild-type (non-silent) human IgG1 Fc region or a variant (silent) Fc region identical to the Fc region of MAB#1. In addition, isotype control antibody MOR03207 was included with either the variant or wild-type human IgG1 Fc region. Assays were performed according to the supplier's instructions using engineered Jurkat cells either expressing Fc.gamma.RIIa_H to mimic the ADCP pathway or Jurkat cells expressing the Fc.gamma.RIIIa, V158 high affinity variant to mimic the ADCC pathway and C5aR expressing CHO cells. A Results from the ADCP reporter bioassay at an IgG concentration of 10 .mu.g/ml. B Results from the ADCC reporter bioassay at an IgG concentration of 10 .mu.g/ml. Data are provided as average fluorescence signal over background.

[0036] FIG. 9: Group mean pharmacokinetic profile of MAB#1 in Han Wistar rats following a single intravenous administration of 10 mg/kg IgG. Data are provided as mean values (IgG concentration over time).+-.standard deviation (S) (n=3).

[0037] FIG. 10: Protein Panel Profiling (3P) results for MAB#1 and MAB#2. The numbers for each antibody represents the obtained binding signal for each antibody and tested protein compared to the binding of isotype negative control antibody MOR03207. Antibodies were tested at a concentration of 10 nM and 100 nM, respectively.

[0038] FIG. 11: Cytokine release by monocyte-derived in vitro-matured M1 and M2 macrophages. IL-10 and IL-12 levels were determined by ELISA after treatment with MAB#1 and incubation with C5a overnight.

DETAILED DESCRIPTION OF THE INVENTION

[0039] The disclosure pertains to a number of human antibodies, which recognize human C5aR.

Definitions

[0040] The term "C5aR" refers to a protein known as C5a anaphylatoxin chemotactic receptor 1 or CD88.

Human C5aR (Uniprot: P2173011-350) (referred to herein as the "D/K variant") has the amino acid sequence of:

TABLE-US-00001 (SEQ ID NO: 1) MDSFNYTTPDYGHYDDKDTLDLNTPVDKTSNTLRVPDILALVIFAVVFLVG VLGNALVVWVTAFEAKRTINAIWFLNLAVADFLSCLALPILFTSIVQHHHW PFGGAACSILPSLILLNMYASILLLATISADRFLLVFKPIWCQNFRGAGLA WIACAVAWGLALLLTIPSFLYRVVREEYFPPKVLCGVDYSHDKRRERAVAI VRLVLGFLWPLLTLTICYTFILLRTWSRRATRSTKTLKVVVAVVASFFIFW LPYQVTGIMMSFLEPSSPTFLLLKKLDSLCVSFAYINCCINPIIYVVAGQG FQGRLRKSLPSLLRNVLTEESVVRESKSFTRSTVDTMAQKTQAV

[0041] Two natural missense mutations of human C5aR are described ((http://www.uniprot.org/uniprot/P21730): Reference SNP (refSNP) Cluster Report: rs4467185 (MAF: 0.03) and Cluster Report: rs11880097 (MAF: 0.03). One mutation is located within the N-terminal extracellular region of human C5aR (Position 2 of SEQ ID NO: 1) resulting in a D to N substitution.

[0042] A human C5aR protein which comprises both natural missense mutations in its sequence (also referred to herein as the "N/N variant") has the amino acid sequence of:

TABLE-US-00002 (SEQ ID NO: 2) MNSFNYTTPDYGHYDDKDTLDLNTPVDKTSNTLRVPDILALVIFAVVFLVG VLGNALVVWVTAFEAKRTINAIWFLNLAVADFLSCLALPILFTSIVQHHHW PFGGAACSILPSLILLNMYASILLLATISADRFLLVFKPIWCQNFRGAGLA WIACAVAWGLALLLTIPSFLYRVVREEYFPPKVLCGVDYSHDKRRERAVAI VRLVLGFLWPLLTLTICYTFILLRTWSRRATRSTKTLKVVVAVVASFFIFW LPYQVTGIMMSFLEPSSPTFLLLNKLDSLCVSFAYINCCINPIIYVVAGQG FQGRLRKSLPSLLRNVLTEESVVRESKSFTRSTVDTMAQKTQAV

Cynomolgus monkey (Macaca fascicularis) C5aR has the amino acid sequence of:

TABLE-US-00003 (SEQ ID NO: 3) MDPFSSTTLDYEHYDGKNVLDSDTPVDKTSNTLRVPDILALVVFAVVFLVG VLGNALVVWVTAFEVKRTINAIWFLNLAVADFLSCLALPILFTSIVQHHHW PFGGTACRILPSLILLNMYASILLLATISADRFLLVFNPIWCQNFRGAGLA WIACAVAWGLALLLTIPSFLYRAVRQEEYSPKVLCGVDYNNDTRRERAVAI VRLVLGFLWPLLTLMICYTFLLLRTWSRRATRSTKTLKVVVAVVASFFIFW LPYQVTGTMMSFLRPSSPTYLQLKKLDSLSISFAYINCCINPVIYVVAGQG FQGRLRKSLPSLLRNVLTEESVVRESKSFTRSTVDTMTEKTQAV

Mouse (Mus musculus) C5aR has the amino acid sequence of

TABLE-US-00004 (SEQ ID NO: 4) MDPIDNSSFEINYDHYGTMAPNIPADGIHLPKRQPGDVAALIIYSVVFLVG VPGNALVVWVTAFEARRAVNAIWFLNLAVADLLSCLALPVLFTTVLNHNYV VYFDATACIVLPSLILLNMYASILLLATISADRFLLVFKPIWCQKVRGTGL AWMACGVAWVLALLLTIPSFVYREAYKDFYSEHTVCGINYGGGSFPKEKAV AILRLMVGFVLPLLTLNICYTFLLLRTWSRKATRSTKTLKVVMAVVICFFI FWLPYQVTGVMIAWLPPSSPTLKRVEKLNSLCVSLAYINCCVNPIIYVMAG QGFHGRLLRSLPSIIRNALSEDSVGRDSKTFTPSTTDTSTRKSQAV

Rat (Rattus norvegicus) C5aR has the amino acid sequence of

TABLE-US-00005 (SEQ ID NO: 5) MDPISNDSSEITYDYSDGTPNPDMPADGVYIPKMEPGDIAALIIYLAVFLV GVTGNALVVWVTAFEAKRTVNAIWFLNLAVADLLSCLALPILFTSIVKHNH WPFGDQACIVLPSLILLNMYSSILLLATISADRFLLVFKPIWCQKFRRPGL AWMACGVTWVLALLLTIPSFVFRRIHKDPYSDSILCNIDYSKGPFFIEKAI AILRLMVGFVLPLLTLNICYTFLLIRTWSRKATRSTKTLKVVMAVVTCFFV FWLPYQVTGVILAWLPRSSSTFQSVERLNSLCVSLAYINCCVNPIIYVMAG QGFHGRLRRSLPSIIRNVLSEDSLGRDSKSFTRSTMDTSTQKSQAV

[0043] The term "C5a" refers to a protein known as Human Complement Component C5a Human C5a (Uniprot: P01031|678-751) has the amino acid sequence of:

TABLE-US-00006 (SEQ ID NO: 6) TLQKKIEEIAAKYKHSVVKKCCYDGACVNNDETCEQRAARISLGPRCIKAF TECCVVASQLRANISHKDMQLGR

[0044] The term "C5L2" refers to a protein known as C5a anaphylatoxin chemotactic receptor 2. Human C5L2 has the amino acid sequence of:

TABLE-US-00007 (SEQ ID NO: 7) MGNDSVSYEYGDYSDLSDRPVDCLDGACLAIDPLRVAPLPLYAAIFLVGVP GNAMVAWVAGKVARRRVGATWLLHLAVADLLCCLSLPILAVPIARGGHWPY GAVGCRALPSIILLTMYASVLLLAALSADLCFLALGPAWWSTVQRACGVQV ACGAAWTLALLLTVPSAIYRRLHQEHFPARLQCVVDYGGSSSTENAVTAIR FLFGFLGPLVAVASCHSALLCWAARRCRPLGTAIVVGFFVCWAPYHLLGLV LTVAAPNSALLARALRAEPLIVGLALAHSCLNPMLFLYFGRAQLRRSLPAA CHWALRESQGQDESVDSKKSTSHDLVSEMEV.

[0045] The term "C3aR" refers to a protein known as C3a anaphylatoxin chemotactic receptor. Human C3aR has the amino acid sequence of:

TABLE-US-00008 (SEQ ID NO: 8) MASFSAETNSTDLLSQPWNEPPVILSMVILSLTFLLGLPGNGLVLWVAGLK MQRTVNTIWFLHLTLADLLCCLSLPFSLAHLALQGQWPYGRFLCKLIPSII VLNMFASVFLLTAISLDRCLVVFKPIWCQNHRNVGMACSICGCIWVVACVM CIPVFVYREIFTTDNHNRCGYKFGLSSSLDYPDFYGDPLENRSLENIVQPP GEMNDRLDPSSFQTNDHPWTVPTVFQPQTFQRPSADSLPRGSARLTSQNLY SNVFKPADVVSPKIPSGFPIEDHETSPLDNSDAFLSTHLKLFPSASSNSFY ESELPQGFQDYYNLGQFTDDDQVPTPLVAITITRLVVGFLLPSVIMIACYS FIVFRMQRGRFAKSQSKTFRVAVVVVAVFLVCWTPYHIFGVLSLLTDPETP LGKTLMSWDHVCIALASANSCFNPFLYALLGKDFRKKARQSIQGILEAAFS EELTRSTHCPSNNVISERNSTTV

[0046] The term "FPR1" refers to a protein known as fMet-Leu-Phe receptor Human FPR1 has the amino acid sequence of:

TABLE-US-00009 (SEQ ID NO: 9) METNSSLPTNISGGTPAVSAGYLFLDIITYLVFAVTFVLGVLGNGLVIWVA GFRMTHTVTTISYLNLAVADFCFTSTLPFFMVRKAMGGHWPFGWFLCKFLF TIVDINLFGSVFLIALIALDRCVCVLHPVWTQNHRTVSLAKKVIIGPWVMA LLLTLPVIIRVTTVPGKTGTVACTFNFSPWTNDPKERINVAVAMLTVRGII RFIIGFSAPMSIVAVSYGLIATKIHKQGLIKSSPPLRVLSFVAAAFFLCWS PYQVVALIATVRIRELLQGMYKEIGIAVDVTSALAFFNSCLNPMLYVFMGQ DFRERLIHALPASLERALTEDSTQTSDTATNSTLPSAEVALQAK

[0047] The term "ChemR23" refers to a protein known as Chemokine-like receptor 1. Human ChemR23 has the amino acid sequence of:

TABLE-US-00010 (SEQ ID NO: 10) MRMEDEDYNTSISYGDEYPDYLDSIVVLEDLSPLEARVTRIFLVVVYSIVC FLGILGNGLVIIIATFKMKKTVNMVWFLNLAVADFLFNVFLPIHITYAAMD YHWVFGTAMCKISNFLLIHNMFTSVFLLTIISSDRCISVLLPVWSQNHRSV RLAYMACMVIWVLAFFLSSPSLVFRDTANLHGKISCFNNFSLSTPGSSSWP THSQMDPVGYSRHMVVTVTRFLCGFLVPVLIITACYLTIVCKLHRNRLAKT KKPFKIIVTIIITFFLCWCPYHTLNLLELHHTAMPGSVFSLGLPLATALAI ANSCMNPILYVFMGQDFKKFKVALFSRLVNALSEDTGHSSYPSHRSFTKMS SMNERTSMNERETGML

[0048] The term "antibody" as used herein refers to a protein comprising at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, which interacts with an antigen. Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region is comprised of three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region is comprised of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR). Each VH and VL is composed of three CDRs and four FR's arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The constant regions of the antibodies may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (CIq) of the classical complement system. The term "antibody" includes for example, monoclonal antibodies, human antibodies, humanized antibodies, camelised antibodies and chimeric antibodies. The antibodies can be of any isotype (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or subclass. Both the light and heavy chains are divided into regions of structural and functional homology.

[0049] The phrase "antibody fragment", as used herein, refers to one or more portions of an antibody that retain the ability to specifically interact with (e.g., by binding, steric hindrance, stabilizing spatial distribution) an antigen. Examples of binding fragments include, but are not limited to, a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; a F(ab)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; a Fd fragment consisting of the VH and CH1 domains; a Fv fragment consisting of the VL and VH domains of a single arm of an antibody; a dAb fragment (Ward et al., (1989) Nature 341:544-546), which consists of a VH domain; and an isolated complementarity determining region (CDR). Furthermore, although the two domains of the Fv fragment, VL and VH, are coded for by separate genes, they can be joined, using recombinant methods, by a synthetic linker that enables them to be made as a single protein chain in which the VL and VH regions pair to form monovalent molecules (known as single chain Fv (scFv); see e.g., Bird et al., (1988) Science 242:423-426; and Huston et al., (1988) Proc. Natl. Acad. Sci. 85:5879-5883). Such single chain antibodies are also intended to be encompassed within the term "antibody fragment". These antibody fragments are obtained using conventional techniques known to those of skill in the art, and the fragments are screened for utility in the same manner as are intact antibodies. Antibody fragments can also be incorporated into single domain antibodies, maxibodies, minibodies, intrabodies, diabodies, triabodies, tetrabodies, v-NAR and bis-scFv (see, e.g., Hollinger and Hudson, (2005) Nature Biotechnology 23:1126-1136). Antibody fragments can be grafted into scaffolds based on polypeptides such as Fibronectin type III (Fn3) (see U.S. Pat. No. 6,703,199, which describes fibronectin polypeptide monobodies). Antibody fragments can be incorporated into single chain molecules comprising a pair of tandem Fv segments (VH-CH1-VH-CH1) which, together with complementary light chain polypeptides, form a pair of antigen-binding sites (Zapata et al., (1995) Protein Eng. 8:1057-1062; and U.S. Pat. No. 5,641,870).

[0050] A "human antibody" or "human antibody fragment", as used herein, is an antibody and antibody fragment having variable regions in which both the framework and CDR regions are from sequences of human origin. Human antibodies can also be isolated from synthetic libraries or from transgenic mice (e.g. Xenomouse) provided the respective system yield in antibodies having variable regions in which both the framework and CDR regions are derived from sequences of human origin. Furthermore, if the antibody contains a constant region, the constant region also is derived from such sequences. Human origin includes, e.g., human germline sequences, or mutated versions of human germline sequences or antibody containing consensus framework sequences derived from human framework sequences analysis, for example, as described in Knappik et al., (2000) J Mol Biol 296:57-86).

[0051] The structures and locations of immunoglobulin variable domains, e.g., CDRs, may be defined using well known numbering schemes, e.g., the Kabat numbering scheme, the Chothia numbering scheme, or a combination of Kabat and Chothia (see, e.g. Sequences of Proteins of Immunological Interest, U.S. Department of Health and Human Services (1991), eds. Kabat et al.; Lazikani et al., (1997) J. Mol. Bio. 273:927-948); Kabat et al., (1991) Sequences of Proteins of Immunological Interest, 5th edit., NIH Publication no. 91-3242 U.S. Department of Health and Human Services; Chothia et al., (1987) J. Mol. Biol. 196:901-917; Chothia et al., (1989) Nature 342:877-883; and Al-Lazikani et al., (1997) J. Mol. Biol. 273:927-948.

[0052] A "humanized antibody" or "humanized antibody fragment" is defined herein as an antibody molecule, which has constant antibody regions derived from sequences of human origin and the variable antibody regions or parts thereof or only the CDRs are derived from another species. For example, a humanized antibody can be CDR-grafted, wherein the CDRs of the variable domain are from a non-human origin, while one or more frameworks of the variable domain are of human origin and the constant domain (if any) is of human origin.

[0053] The term "chimeric antibody" or "chimeric antibody fragment" is defined herein as an antibody molecule, which has constant antibody regions derived from, or corresponding to, sequences found in one species and variable antibody regions derived from another species. Preferably, the constant antibody regions are derived from, or corresponding to, sequences found in humans, and the variable antibody regions (e.g. VH, VL, CDR or FR regions) are derived from sequences found in a non-human animal, e.g. a mouse, rat, rabbit or hamster.

[0054] The term "isolated antibody" refers to an antibody or antibody fragment that is substantially free of other antibodies or antibody fragments having different antigenic specificities. Moreover, an isolated antibody or antibody fragment may be substantially free of other cellular material and/or chemicals. Thus, in some aspects, antibodies provided are isolated antibodies, which have been separated from antibodies with a different specificity. An isolated antibody may be a monoclonal antibody. An isolated antibody may be a recombinant monoclonal antibody. An isolated antibody that specifically binds to an epitope, isoform or variant of a target may, however, have cross-reactivity to other related antigens, e.g., from other species (e.g., species homologs).

[0055] The term "recombinant antibody", as used herein, includes all antibodies that are prepared, expressed, created or segregated by means not existing in nature. For example, antibodies isolated from a host cell transformed to express the antibody, antibodies selected and isolated from a recombinant, combinatorial human antibody library, and antibodies prepared, expressed, created or isolated by any other means that involve splicing of all or a portion of a human immunoglobulin gene, sequences to other DNA sequences or antibodies isolated from an animal (e.g., a mouse) that is transgenic or transchromosomal for human immunoglobulin genes or a hybridoma prepared therefrom. Preferably, such recombinant antibodies have variable regions in which the framework and CDR regions are derived from human germline immunoglobulin sequences. In certain embodiments, however, such recombinant human antibodies can be subjected to in vitro mutagenesis (or, when an animal transgenic for human Ig sequences is used, in vivo somatic mutagenesis) and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are sequences that, while derived from and related to human germline VH and VL sequences, may not naturally exist within the human antibody germline repertoire in vivo. A recombinant antibody may be a monoclonal antibody. In an embodiment, the antibodies and antibody fragment disclosed herein are isolated from the HuCAL library (Rothe et al, J. Mol. Biol. (2008) 376, 1182-1200).

[0056] As used herein, an antibody "binds specifically to", "specifically binds to", is "specific to/for" or "specifically recognizes" an antigen, such as human C5aR, if such antibody is able to discriminate between such antigen and one or more reference antigen(s), since binding specificity is not an absolute, but a relative property. For example, a standard ELISA assay can be carried out. The scoring may be carried out by standard color development (e.g. secondary antibody with horseradish peroxide and tetramethyl benzidine with hydrogen peroxide). The reaction in certain wells is scored by the optical density, for example, at 450 nm. Typical background (=negative reaction) may be 0.1 OD; typical positive reaction may be 1 OD. This means the difference positive/negative can be more than 10-fold. Typically, determination of binding specificity is performed by using not a single reference antigen, but a set of about three to five unrelated antigens, such as milk powder, BSA, transferrin or the like.

[0057] As used herein, the term "affinity" refers to the strength of interaction between the polypeptide and its target at a single site. Within each site, the binding region of the polypeptide interacts through weak non-covalent forces with its target at numerous sites; the more interactions, the stronger the affinity.

[0058] The term "K.sub.D", as used herein, refers to the dissociation constant, which is obtained from the ratio of k.sub.off to K.sub.on (i.e. k.sub.off/k.sub.on) and is expressed as a molar concentration (M). K.sub.D values for antigen binding moieties like e.g. monoclonal antibodies can be determined using methods well established in the art. Methods for determining the K.sub.D of an antigen binding moiety like e.g. a monoclonal antibody are SET (solution equilibrium titration) or surface plasmon resonance using a biosensor system such as a Biacore.RTM. system. In the present disclosure an antibody specific for C5aR typically has a dissociation rate constant (K.sub.D) (k.sub.off/k.sub.on) of less than 5.times.10.sup.-2M, less than 10.sup.-2M, less than 5.times.10.sup.-3M, less than 10.sup.-3M, less than 5.times.10.sup.-4M, less than 10.sup.-4M, less than 5.times.10.sup.-5M, less than 10.sup.-5M, less than 5.times.10.sup.-6M, less than 10.sup.-6M, less than 5.times.10.sup.-7M, less than 10.sup.-7M, less than 5.times.10.sup.-6M, less than 10.sup.-8M, less than 5.times.10.sup.-9M, less than 10.sup.-9M, less than 5.times.10.sup.-10M, less than 10.sup.-10M, less than 5.times.10.sup.-11M, less than 10.sup.-11M, less than 5.times.10.sup.-12M, less than 10.sup.-12M, less than 5.times.10.sup.-13M, less than 10.sup.-13M, less than 5.times.10.sup.-14M, less than 10.sup.-14M, less than 5.times.10.sup.-15M, or less than 10.sup.-15M or lower.

[0059] The term "epitope" includes any proteinacious region which is specifically recognized by an antibody or antibody fragment thereof or otherwise interacts with a molecule. Generally, epitopes are of chemically active surface groupings of molecules such as amino acids or carbohydrate or sugar side chains and generally may have specific three-dimensional structural characteristics, as well as specific charge characteristics. As will be appreciated by one of skill in the art, practically anything to which an antibody can specifically bind could be an epitope.

[0060] "Compositions" of the present disclosure may be used for therapeutic or prophylactic applications. The present disclosure, therefore, includes a pharmaceutical composition containing an antibody or antibody fragment as disclosed herein and a pharmaceutically acceptable carrier or excipient therefor. In a related aspect, the present disclosure provides a method for treating cancer. Such method contains the steps of administering to a subject in need thereof an effective amount of the pharmaceutical composition that contains an antibody or antibody fragment as described herein.

[0061] The present disclosure provides therapeutic methods comprising the administration of a therapeutically effective amount of an antibody or antibody fragment as disclosed herein to a subject in need of such treatment. A "therapeutically effective amount" or "effective amount", as used herein, refers to the amount of a C5aR antibody necessary to elicit the desired biological response. In accordance with the subject disclosure, the therapeutic effective amount is the amount of a C5aR antibody necessary to treat and/or prevent a disease.

[0062] "Administered" or "administration" includes but is not limited to delivery of a drug by an injectable form, such as, for example, an intravenous, intramuscular, intradermal or subcutaneous route or mucosal route, for example, as a nasal spray or aerosol for inhalation or as an ingestible solution, capsule or tablet. Preferably, the administration is by an injectable form.

[0063] As used herein, "treatment", "treat" or "treating" and the like refers to clinical intervention in an attempt to alter the natural course of a disease in the subject being treated, and can be performed either for prophylaxis or during the course of clinical pathology. Desirable effects of treatment include, but are not limited to, preventing occurrence or recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis. In some embodiments, antibodies or antibody fragments according to the preset disclosure are used to delay development of a disease or to slow the progression of a disease.

[0064] The term "effector function" refers to those biological activities attributable to the Fc region of an antibody, which vary with the antibody isotype. Non-limiting examples of antibody effector functions include C1q binding and complement dependent cytotoxicity (CDC); Fc receptor binding and antibody-dependent cell-mediated cytotoxicity (ADCC) and/or antibody-dependent cellular phagocytosis (ADCP); down regulation of cell surface receptors (e.g. B cell receptor); and B cell activation.

[0065] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a form of cytotoxicity in which antibodies bound onto Fc receptors (FcRs) present on certain cytotoxic cells (e.g. NK cells, neutrophils, and macrophages) enable these cytotoxic effector cells to bind specifically to an antigen-bearing target cell and subsequently kill the target cell with cytotoxins. The primary cells for mediating ADCC, NK cells, express Fc.gamma.RIII only, whereas monocytes express Fc.gamma.RI, Fc.gamma.RII, and Fc.gamma.RIII.

[0066] "Complement-dependent cytotoxicity" or "CDC" refers to the lysis of a target cell 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 (of the appropriate subclass) of the present disclosure, which are bound to their cognate antigen.

[0067] "Antibody-dependent cellular phagocytosis" or "ADCP" refers to a mechanism of elimination of antibody-coated target cells by internalization by phagocytic cells, such as macrophages or dendritic cells.

[0068] `Preventing` or `prevention` refers to a reduction in risk of acquiring or developing a disease (i.e. causing at least one of the clinical symptoms of the disease not to develop in a subject that may be exposed to a disease-causing agent, or predisposed to the disease in advance of disease onset). "Prevention" also refers to methods which aim to prevent the onset of a disease or its symptoms or which delay the onset of a disease or its symptoms.

[0069] "Subject" or "species" or as used in this context refers to any mammal, including rodents, such as mouse or rat, and primates, such as cynomolgus monkey (Macaca fascicularis), rhesus monkey (Macaca mulatta) or humans (Homo sapiens). Preferably, the subject is a primate, most preferably a human.

[0070] Throughout this specification, unless the context requires otherwise, the words "comprise", "have" and "include" and their respective variations such as "comprises", "comprising", "has", "having", "includes" and "including" will be understood to imply the inclusion of a stated element or integer or group of elements or integers but not the exclusion of any other element or integer or group of elements or integers.

[0071] The terms "engineered" or "modified" as used herein includes manipulation of nucleic acids or polypeptides by synthetic means (e.g. by recombinant techniques, in vitro peptide synthesis, by enzymatic or chemical coupling of peptides or some combination of these techniques). Preferably, the antibodies or antibody fragments according to the present disclosure are engineered or modified to improve one or more properties, such as antigen binding, stability, half-life, effector function, immunogenicity, safety and the like.

[0072] "Variant" as used herein refers to a polypeptide that differs from a reference polypeptide by one or more modifications for example amino acid substitutions, insertions or deletions.

[0073] The term "amino acid mutation" as used herein is meant to encompass amino acid substitutions, deletions, insertions, and modifications. Any combination of substitution, deletion, insertion, and modification can be made as long as the final construct possesses the desired characteristics, e.g., reduced binding to an Fc receptor. Amino acid sequence deletions and insertions include N- and/or C-terminal deletions and insertions of amino acid residues. Particular amino acid mutations are amino acid substitutions. Amino acid substitutions include replacement by non-naturally occurring amino acids or by naturally occurring amino acid derivatives of the twenty standard amino acids. Amino acid mutations can be generated using genetic or chemical methods well known in the art. Genetic methods may include site-directed mutagenesis, PCR, gene synthesis and the like. It is contemplated that methods of altering the side chain group of an amino acid residue by methods other than genetic engineering, such as chemical modification, may also be useful. Various designations may be used herein to indicate the same amino acid mutation. For example, a substitution from glyince at position 327 of the Fc region to alanine can be indicated as 237A, G337, G337A, or Gly329Ala.

[0074] The term "EC.sub.50" as used herein, refers to the concentration of an antibody or antibody fragment or ligand, which induces a response in an assay half way between the baseline and maximum. It therefore represents the antibody or ligand concentration at which 50% of the maximal effect is observed

[0075] The term "IC.sub.50" as used herein, refers to the concentration of an antibody or antibody fragment that inhibits a response in an assay half way between the maximal response and the baseline. It represents the antibody concentration that reduces a given response by 50%.

[0076] The terms "inhibition" or "inhibit" or "reduction" or "reduce" or "neutralization" or "neutralize" refer to a decrease or cessation of any phenotypic characteristic (such as binding or a biological activity or function) or to the decrease or cessation in the incidence, degree, or likelihood of that characteristic. The "inhibition", "reduction" or "neutralization" needs not to be complete as long as it is detectable using an appropriate assay. In some embodiments, by "reduce" or "inhibit" or "neutralize" is meant the ability to cause a decrease of 20% or greater. In another embodiment, by "reduce" or "inhibit" or "neutralize" is meant the ability to cause a decrease of 50% or greater. In yet another embodiment, by "reduce" or "inhibit" or "neutralize" is meant the ability to cause an overall decrease of 75%, 85%, 90%, 95%, or greater.

[0077] The term "antagonistic" antibody as used herein refers to an antibody or antibody fragment that interacts with an antigen and partially or fully inhibits or neutralizes a biological activity or function or any other phenotypic characteristic of a target antigen.

[0078] A "wild-type" protein is a version or variant of the protein as it is found in nature. An amino acid sequence of a wildtype protein, e.g., a Fc region of an human IgG1 antibody, is the amino acid sequence of the protein as it occurs in nature. Due to allotypic differences, there can be more than one amino acid sequence for a wildtype protein. For example, there are several allotypes of naturally occurring human IGg1 heavy chain constant regions (see, e.g., Jeffries et al. (2009) mAbs 1:1).

[0079] The "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain. The Fc region of an immunoglobulin generally comprises two constant domains, a CH2 domain and a CH3 domain. Although the boundaries of the Fc region of an IgG heavy chain might vary slightly, the human IgG heavy chain Fc region is usually defined to extend from Cys226, or from Pro230, to the C-terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. Unless otherwise specified herein, numbering of amino acid residues in the Fc region is according to the EU numbering system, also called the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991.

Embodiments

[0080] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises [0081] a) the HCDR1 region comprising the amino acid sequence of SEQ ID NO: 27, the HCDR2 region comprising the amino acid sequence of SEQ ID NO: 28, the HCDR3 region comprising the amino acid sequence of SEQ ID NO: 29, the LCDR1 region comprising the amino acid sequence of SEQ ID NO: 32, the LCDR2 region comprising the amino acid sequence of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34, or [0082] b) the HCDR1 region comprising the amino acid sequence of SEQ ID NO: 27, the HCDR2 region comprising the amino acid sequence of SEQ ID NO: 39, the HCDR3 region comprising the amino acid sequence of SEQ ID NO: 40, the LCDR1 region comprising the amino acid sequence of SEQ ID NO: 32, the LCDR2 region comprising the amino acid sequence of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34.

[0083] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises [0084] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0085] b) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0086] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises [0087] a) the HCDR1 region comprising the amino acid sequence of SEQ ID NO: 30, the HCDR2 region comprising the amino acid sequence of SEQ ID NO: 31, the HCDR3 region comprising the amino acid sequence of SEQ ID NO: 29, the LCDR1 region comprising the amino acid sequence of SEQ ID NO: 32, the LCDR2 region comprising the amino acid sequence of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34, or [0088] b) the HCDR1 region comprising the amino acid sequence of SEQ ID NO: 30, the HCDR2 region comprising the amino acid sequence of SEQ ID NO: 41, the HCDR3 region comprising the amino acid sequence of SEQ ID NO: 40, the LCDR1 region comprising the amino acid sequence of SEQ ID NO: 32, the LCDR2 region comprising the amino acid sequence of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34.

[0089] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises [0090] a) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0091] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0092] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises [0093] a) the HCDR1 region comprising the amino acid sequence of SEQ ID NO: 27, the HCDR2 region comprising the amino acid sequence of SEQ ID NO: 28, the HCDR3 region comprising the amino acid sequence of SEQ ID NO: 29, the LCDR1 region comprising the amino acid sequence of SEQ ID NO: 32, the LCDR2 region comprising the amino acid sequence of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34, or [0094] b) the HCDR1 region comprising the amino acid sequence of SEQ ID NO: 30, the HCDR2 region comprising the amino acid sequence of SEQ ID NO: 31, the HCDR3 region comprising the amino acid sequence of SEQ ID NO: 29, the LCDR1 region comprising the amino acid sequence of SEQ ID NO: 32, the LCDR2 region comprising the amino acid sequence of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34, or [0095] c) the HCDR1 region comprising the amino acid sequence of SEQ ID NO: 27, the HCDR2 region comprising the amino acid sequence of SEQ ID NO: 39, the HCDR3 region comprising the amino acid sequence of SEQ ID NO: 40, the LCDR1 region comprising the amino acid sequence of SEQ ID NO: 32, the LCDR2 region comprising the amino acid sequence of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34, or [0096] d) the HCDR1 region comprising the amino acid sequence of SEQ ID NO: 30, the HCDR2 region comprising the amino acid sequence of SEQ ID NO: 41, the HCDR3 region comprising the amino acid sequence of SEQ ID NO: 40, the LCDR1 region comprising the amino acid sequence of SEQ ID NO: 32, the LCDR2 region comprising the amino acid sequence of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34.

[0097] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises [0098] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0099] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0100] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0101] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0102] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0103] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0104] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0105] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0106] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR comprising 6 CDRs defined by Kabat of any one of the antibodies disclosed in Table 1 or Table 2.

[0107] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR comprising 6 CDRs defined by Chothia of any one of the antibodies disclosed in Table 1 or Table 2.

[0108] In an embodiment of the present disclosure, the isolated antibody or antibody fragment is a monoclonal antibody or antibody fragment. In an embodiment of the present disclosure, the isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment. In another embodiment of the present disclosure, the isolated antibody or antibody fragment is recombinant antibody or antibody fragment. In an embodiment of the present disclosure, the isolated antibody or antibody fragment is of the IgG isotype. In an embodiment of the present disclosure, the isolated antibody or antibody fragment is of the IgG1 class. In another embodiment of the present disclosure, the isolated antibody or antibody fragment does not substantially induce effector function in vitro.

[0109] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises [0110] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34 and further comprises the VH of SEQ ID NO: 35 or the VL of SEQ ID NO: 36, or [0111] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34 and further comprises the VH of SEQ ID NO: 35 or the VL of SEQ ID NO: 36, or [0112] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34 and further comprises the VH of SEQ ID NO: 42 or the VL of SEQ ID NO: 43, or [0113] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34 and further comprises the VH of SEQ ID NO: 42 or the VL of SEQ ID NO: 43.

[0114] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises [0115] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0116] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43.

[0117] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36.

[0118] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43.

[0119] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38.

[0120] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45.

[0121] In an embodiment, the present disclosure refers to an isolated_antibody or antibody fragment specific for human C5aR comprising the variable heavy chain (VH) and the variable light chain (VL) of any one of the antibodies disclosed in Table 1 or Table 2.

[0122] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR comprising the heavy chain (HC) and the light chain (LC) of any one of the antibodies disclosed in Table 1 or Table 2.

[0123] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34 and further comprises the VH of SEQ ID NO: 35 or the VL of SEQ ID NO: 36

[0124] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34 and further comprises the VH of SEQ ID NO: 42 or the VL of SEQ ID NO: 43

[0125] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34 and further comprises the VH of SEQ ID NO: 35 or the VL of SEQ ID NO: 36

[0126] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34 and further comprises the VH of SEQ ID NO: 42 or the VL of SEQ ID NO: 43.

[0127] In an embodiment of the present disclosure, the isolated antibody or antibody fragment is a monoclonal antibody or antibody fragment. In an embodiment of the present disclosure, the isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment. In another embodiment of the present disclosure, the isolated antibody or antibody fragment is recombinant antibody or antibody fragment.

[0128] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises [0129] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0130] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or

[0131] a VH and a VL that has at least at 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or SEQ ID NO: 42 and to the VL of SEQ ID NO: 36 or SEQ ID NO: 43.

[0132] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36, or a VH and a VL that has at least at 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 and to the VL of SEQ ID NO: 36.

[0133] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43, or a VH and a VL that has at least at 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 42 and to the VL of SEQ ID NO: 43.

[0134] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises [0135] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0136] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or

[0137] a HC and a LC that has at least at 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or SEQ ID NO: 44 and to the LC of SEQ ID NO:38 or SEQ ID NO: 45.

[0138] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or a HC and a LC that has at least at 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 and to the LC of SEQ ID NO: 38.

[0139] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or a HC and a LC that has at least at 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 44 and to the LC of SEQ ID NO: 45.

[0140] In an embodiment of the present disclosure, the isolated antibody or antibody fragment is a monoclonal antibody or antibody fragment. In an embodiment of the present disclosure, the isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment. In another embodiment of the present disclosure, the isolated antibody or antibody fragment is recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or an antibody fragment is a human or humanized antibody or antibody fragment. In one embodiment, said isolated antibody or an antibody fragment is a human antibody or antibody fragment.

[0141] In an embodiment, the isolated antibody or antibody fragment specific for human C5aR of the present disclosure is a recombinant or synthetic antibody or antibody fragment. In a further embodiment, the isolated antibody or antibody fragment specific for C5aR according to the present disclosure is an isolated recombinant monoclonal antibody or antibody fragment. In a further embodiment, the isolated antibody or antibody fragment specific for C5aR according to the present disclosure is an isolated recombinant monoclonal human antibody or antibody fragment.

[0142] In an embodiment, the isolated antibody or antibody fragment specific for C5aR according to the present disclosure is of the IgG isotype. In another embodiment, said isolated antibody or antibody fragment is of the IgG1 class. In another embodiment, said isolated antibody or antibody fragment is of the human IgG1 class.

Nucleic Acids

[0143] In an embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated said antibody or antibody fragment comprise [0144] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0145] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0146] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0147] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0148] In another embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding the heavy chain sequence and light chain sequence of an isolated antibody or antibody fragment specific for human C5aR, wherein the nucleic acid sequence or a plurality of nucleic acid sequences comprises [0149] a) the HCDR1 region of SEQ ID NO: 46, the HCDR2 region of SEQ ID NO: 47, the HCDR3 region of SEQ ID NO: 48, the LCDR1 region of SEQ ID NO: 51, the LCDR2 region of SEQ ID NO: 52 and the LCDR3 region of SEQ ID NO: 53, or [0150] b) the HCDR1 region of SEQ ID NO: 49, the HCDR2 region of SEQ ID NO: 50, the HCDR3 region of SEQ ID NO: 48, the LCDR1 region of SEQ ID NO: 51, the LCDR2 region of SEQ ID NO: 52 and the LCDR3 region of SEQ ID NO: 53, or [0151] c) the HCDR1 region of SEQ ID NO: 58, the HCDR2 region of SEQ ID NO: 59, the HCDR3 region of SEQ ID NO: 60, the LCDR1 region of SEQ ID NO: 63, the LCDR2 region of SEQ ID NO: 64 and the LCDR3 region of SEQ ID NO: 65, or [0152] d) the HCDR1 region of SEQ ID NO: 61, the HCDR2 region of SEQ ID NO: 62, the HCDR3 region of SEQ ID NO: 60, the LCDR1 region of SEQ ID NO: 63, the LCDR2 region of SEQ ID NO: 64 and the LCDR3 region of SEQ ID NO: 65.

[0153] In another embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding the heavy chain sequence and light chain sequence of an isolated antibody or antibody fragment specific for human C5aR, wherein the nucleic acid sequence or a plurality of nucleic acid sequences comprise [0154] a) the HCDR1 region of SEQ ID NO: 70, the HCDR2 region of SEQ ID NO: 71, the HCDR3 region of SEQ ID NO: 72, the LCDR1 region of SEQ ID NO: 75, the LCDR2 region of SEQ ID NO: 76 and the LCDR3 region of SEQ ID NO: 77, or [0155] b) the HCDR1 region of SEQ ID NO: 73, the HCDR2 region of SEQ ID NO: 74, the HCDR3 region of SEQ ID NO: 72, the LCDR1 region of SEQ ID NO: 75, the LCDR2 region of SEQ ID NO: 76 and the LCDR3 region of SEQ ID NO: 77, or [0156] c) the HCDR1 region of SEQ ID NO: 82, the HCDR2 region of SEQ ID NO: 83, the HCDR3 region of SEQ ID NO: 84, the LCDR1 region of SEQ ID NO: 87, the LCDR2 region of SEQ ID NO: 88 and the LCDR3 region of SEQ ID NO: 89, or [0157] d) the HCDR1 region of SEQ ID NO: 85, the HCDR2 region of SEQ ID NO: 86, the HCDR3 region of SEQ ID NO: 84, the LCDR1 region of SEQ ID NO: 87, the LCDR2 region of SEQ ID NO: 88 and the LCDR3 region of SEQ ID NO: 89.

[0158] In an embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein said nucleic acid sequence or plurality of nucleic acid sequences comprise the VH of SEQ ID NO: 54 and the VL of SEQ ID NO: 55, or a VH and a VL that has at least 60%, at least 70%, at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 54 and/or the VL of SEQ ID NO: 55.

[0159] In an embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein said nucleic acid sequence or plurality of nucleic acid sequences comprise the VH of SEQ ID NO: 66 and the VL of SEQ ID NO: 67, or a VH and a VL that has at least 60%, at least 70%, at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 66 and/or the VL of SEQ ID NO: 67.

[0160] In an embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein said nucleic acid sequence or plurality of nucleic acid sequences comprise the VH of SEQ ID NO: 78 and the VL of SEQ ID NO: 79, or a VH and a VL that has at least 60%, at least 70%, at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 78 and/or the VL of SEQ ID NO: 79.

[0161] In an embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein said nucleic acid sequence or plurality of nucleic acid sequences comprise the VH of SEQ ID NO: 90 and the VL of SEQ ID NO: 91, or a VH and a VL that has at least 60%, at least 70%, at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 90 and/or the VL of SEQ ID NO: 91.

[0162] In an embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein said nucleic acid sequence or plurality of nucleic acid sequences comprise the HC of SEQ ID NO: 56 and the LC of SEQ ID NO: 57, or a HC and a LC that has at least 60%, at least 70%, at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 56 and/or the LC of SEQ ID NO: 57.

[0163] In an embodiment, the present disclosure refers to an nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein said nucleic acid sequence or plurality of nucleic acid sequences comprise the HC of SEQ ID NO: 68 and the LC of SEQ ID NO: 69, or a HC and a LC that has at least 60%, at least 70%, at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 68 and/or the LC of SEQ ID NO: 69.

[0164] In an embodiment, the present disclosure refers to an nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein said nucleic acid sequence or plurality of nucleic acid sequences comprise the HC of SEQ ID NO: 80 and the LC of SEQ ID NO: 81, or a HC and a LC that has at least 60%, at least 70%, at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 80 and/or the LC of SEQ ID NO: 81.

[0165] In an embodiment, the present disclosure refers to an nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein said nucleic acid sequence or plurality of nucleic acid sequences comprise the HC of SEQ ID NO: 92 and the LC of SEQ ID NO: 93, or a HC and a LC that has at least 60%, at least 70%, at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 92 and/or the LC of SEQ ID NO: 93.

[0166] In an embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein said nucleic acid sequence or plurality of nucleic acid sequences comprise the HC of SEQ ID NO: 56 and the LC of SEQ ID NO: 57.

[0167] In an embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein said nucleic acid sequence or plurality of nucleic acid sequences comprise the HC of SEQ ID NO: 68 and the LC of SEQ ID NO: 69.

[0168] In an embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein said nucleic acid sequence or plurality of nucleic acid sequences comprise the HC of SEQ ID NO: 80 and the LC of SEQ ID NO: 81.

[0169] In an embodiment, the present disclosure refers to an nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an antibody or antibody fragment specific for human C5aR disclosed herein, wherein said nucleic acid sequence or plurality of nucleic acid sequences comprise the HC of SEQ ID NO: 92 and the LC of SEQ ID NO: 93.

[0170] In another embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or fragment specific for human C5aR, wherein the nucleic acid sequence or the plurality of nucleic acid sequences comprise the VH and VL of any one of the antibodies disclosed in Table 1 or Table 2.

[0171] In an embodiment, the present disclosure provides an isolated antibody or antibody fragment specific for human C5aR, encoded by any one of the nucleic acid sequences or plurality of nucleic acid sequences disclosed in Table 1 or Table 2.

[0172] In an embodiment, the present disclosure provides a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding any one of the isolated antibodies or antibody fragments disclosed in Table 1 or Table 2.

[0173] In an embodiment, the present disclosure provides a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding any one of the isolated antibodies or antibody fragments specific for human C5aR according to the present disclosure.

[0174] In another embodiment, the present disclosure refers to a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR, wherein the nucleic acid sequence or the plurality of nucleic acid sequences comprises a HC and a LC of any one of the antibodies or antibody fragments disclosed in Table 1 or Table 2.

[0175] In an embodiment, said nucleic acid composition and/or said nucleic acid sequence and/or plurality of nucleic acid sequences are isolated.

Vectors

[0176] In an embodiment, the present disclosure provides a vector composition comprising a vector or a plurality of vectors comprising a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR according to the present disclosure.

[0177] In an embodiment, the present disclosure provides a vector composition comprising a vector or a plurality of vectors comprising a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding any one of the isolated antibodies or antibody fragments specific for human C5aR disclosed in Tables 1 or Table 2.

[0178] In an embodiment, the present disclosure provides a vector composition comprising a vector or a plurality of vectors comprising a nucleic acid sequence or a plurality of nucleic acid sequences disclosed in Table 1 or Table 2.

[0179] In an embodiment, said vector composition and/or vector and/or plurality of vectors are isolated.

Host Cells

[0180] In an embodiment, the present disclosure provides a host cell comprising a vector composition comprising a vector or a plurality of vectors comprising a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for human C5aR according to the present disclosure.

[0181] In an embodiment, the present disclosure refers to a host cell comprising a vector composition comprising a vector or a plurality of vectors comprising a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an isolated antibody or antibody fragment specific for C5aR disclosed in Table 1 or Table 2.

[0182] In an embodiment, the host cell according to the present disclosure is able to express the isolated antibody or antibody fragment specific for human C5aR encoded by the vector composition or the nucleic acid composition.

[0183] In a further embodiment, the host cell is an isolated host cell. In a further embodiment, said host cell is a mammalian cell. In an embodiment, said mammalian cell is a human cell. In another embodiment, said mammalian cell is a CHO cell. In an embodiment, said cell is a HEK cell. In another embodiment, said cell is a PERC.6 cell. In an embodiment, said cell is a HKB11 cell.

[0184] The skilled man will realize that the nucleic acid sequence or the plurality of nucleic acid sequences encoding the heavy and/or light chain of an antibody or antibody fragment of the present disclosure can be cloned into different vectors or into the same vector.

[0185] The vectors can be introduced into the appropriate host cells such as prokaryotic (e.g., bacterial) or eukaryotic (e.g., yeast or mammalian) cells by methods well known in the art (see e.g., "Current Protocol in Molecular Biology", Ausubel et al. (eds.), Greene Publishing Assoc. and John Wiley Interscience, New York, 1989 and 1992). Numerous cloning vectors are known to those of skill in the art, and the selection of an appropriate cloning vector is a matter of choice. The gene can be placed under the control of a promoter, ribosome binding site (for bacterial expression) and, optionally, an operator (collectively referred to herein as "control" elements), so that the nucleic acid sequence encoding the desired protein is transcribed into RNA in the host cell transformed by a vector containing this expression construction. The coding sequence may or may not contain a signal peptide or leader sequence. Upon expression in host cells, the antibodies or antibody fragments of the present disclosure are obtained. These steps can be achieved in different ways, as will be known by the person skilled in the art. In general, such steps typically include transforming or transfecting a suitable host cell with a nucleic acid composition or vector composition or an infectious particle, which encodes the antibody, or antibody fragments. Further, such steps typically include culturing said host cells under conditions suitable for the proliferation (multiplication, growth) of said host cells and a culturing step under conditions suitable for the production (expression, synthesis) of the encoded antibody or antibody fragment. The culturing of host cells under conditions suitable for proliferation or expression is typically accomplished in the presence of media comprising components suitable for cell growth or induction of expression. In particular, embodiments, the methods for the production of the antibodies or antibody fragments of the present disclosure further comprise the step of isolating and purifying the produced antibody or antibody fragment from the host cells or medium. If the expression system secretes the protein into growth media, the protein can be purified directly from the media. If the protein is not secreted, it is isolated from cell lysates or recovered from the cell membrane fraction. The selection of the appropriate growth conditions and recovery methods are within the skill of the art. The antibody or antibody fragment of the present disclosure can then be purified by a number of techniques as known to the person skilled in the art.

[0186] In an embodiment, the present disclosure refers to a method of producing an isolated antibody or antibody fragment specific for human C5aR of any of the antibodies disclosed in Table 1 or Table 2. In an embodiment, a method of producing an isolated antibody or antibody fragment according to the present disclosure is provided, wherein the method comprises culturing a host cell comprising a vector composition comprising a vector or a plurality of vectors comprising a nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding an antibody or antibody fragment according to the present disclosure, under conditions suitable for expression of the antibody or antibody fragment, and isolating the antibody or antibody fragment from the host cell or host cell culture medium. An antibody or antibody fragment isolated as described herein may be purified techniques know in the art, such as high performance liquid chromatography (HPLC), ion exchange chromatography, gel electrophoresis, affinity chromatography, size exclusion chromatography, and the like. The conditions used to purify a particular antibody or antibody fragment will depend, in part, on factors such as net charge, hydrophobicity, hydrophilicity etc., and will be apparent to those having skill in the art. For affinity chromatography purification an antibody, ligand, receptor or antigen can be used to which the antibody or antibody fragment binds. For example, for affinity chromatography purification of antibody or antibody fragment according to the present disclosure, a matrix with protein A or protein G may be used. The purity of an antibody or antibody fragment can be determined by any of a variety of well-known analytical methods including gel electrophoresis, high-pressure liquid chromatography, and the like.

Specificity

[0187] In an embodiment, the present disclosure pertains to an isolated antibody or antibody fragment specific for human C5aR disclosed in Table 1 or Table 2. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure is specific for human C5aR.

[0188] In an embodiment, the isolated antibody or antibody fragment according to the present disclosure is specific for human C5aR encoded by the amino acid sequence of SEQ ID NO: 1 and/or SEQ ID NO: 2. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure is specific for a polypeptide comprising the amino acid sequence of SEQ ID NO: 1 and/or SEQ ID NO: 2. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure is specific for a polypeptide consisting of the amino acid sequence of SEQ ID NO: 1 and/or SEQ ID NO: 2.

[0189] In an embodiment, the isolated antibody or antibody fragment according to the present disclosure specifically binds to the extracellular region human C5aR. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure specifically binds to the N-terminal extracellular region of human C5aR. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure specifically binds to the N-terminal extracellular region of human C5aR, wherein the N-terminal extracellular region comprises the amino acid sequence of SEQ ID NO: 13. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure specifically binds to the N-terminus of human C5aR comprising the amino acid sequence of SEQ ID NO: 13. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure specifically binds to a peptide comprising the amino acid sequence of SEQ ID NO: 13. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure specifically binds to a peptide consisting of the amino acid sequence of SEQ ID NO: 13.

[0190] In an embodiment, the said isolated antibody or antibody fragment specific for human C5aR, comprises [0191] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0192] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0193] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0194] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0195] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0196] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0197] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0198] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0199] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0200] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0201] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a human, humanized or chimeric antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is an isolated recombinant human monoclonal antibody or antibody fragment.

Species Cross-Reactivity

[0202] In further embodiments, the isolated antibody or antibody fragment according to the present disclosure is cross-reactive to cynomolgus monkey (cynomolgus) C5aR. In another embodiment, the isolated antibody or antibody fragment according to the present disclosure is specific for human C5aR and cynomolgus C5aR.

[0203] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment cross-reactively binds to cynomolgus C5aR. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure specifically binds to the extracellular region of human C5aR and cynomolgus C5aR. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure binds to the N-terminal extracellular region of human C5aR and cynomolgus C5aR. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure binds to the N-terminal extracellular region of human C5aR and cynomolgus C5aR, wherein the N-terminal extracellular region of C5aR comprises the amino acid sequence of SEQ ID NO: 13 or SEQ ID NO: 14.

[0204] In yet another embodiment, the isolated antibody or antibody fragment according to the present disclosure does not bind to rodent C5aR, such as mouse or rat C5aR.

[0205] In an embodiment, the antibody or antibody fragment according to the present disclosure binds to a peptide comprising the amino acid sequence of SEQ ID NO: 13 and/or SEQ ID NO: 14.

[0206] In an embodiment, the antibody or antibody fragment according to the present disclosure binds to a peptide consisting of the amino acid sequence of SEQ ID NO: 13 and/or SEQ ID NO: 14.

[0207] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR and cynomolgus C5aR, comprises [0208] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0209] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0210] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0211] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0212] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0213] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0214] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0215] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0216] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0217] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0218] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

Monovalent Affinity for C5aR Peptides

[0219] In further embodiments, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment has a monovalent affinity for a human C5aR peptide comprising SEQ ID NO: 13 with a K.sub.D of 100 nM or less, such as 90 nM or less, 80 nM or less, 70 nM or less, 60 nM or less, 50 nM or less, 40 nM or less, 30 nM or less, 20 nM or less, 10 nM or less, 5 nM or 1 nM less.

[0220] In an embodiment, said monovalent affinity is determined in IgG format. In certain embodiments, said monovalent affinity is determined by surface plasmon resonance (SPR) as described herein in Example 4.

[0221] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR, comprises [0222] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0223] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0224] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0225] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0226] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0227] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0228] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0229] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0230] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0231] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0232] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0233] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

Apparent Affinity (Bivalent) for C5aR Peptides

[0234] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody has an apparent affinity for a human C5aR peptide comprising SEQ ID NO: 13 with a K.sub.D of 1 nM or less, such as 0.5 nM or less, 0.4 nM or less, 0.3 nM or less, 0.2 nM or less, 0.1 nM or less, 90 pM or less, 80 pM or less, 70 pM or less, 60 pM or less, 50 pM or less, 40 pM or less, 30 pM or less, 20 pM or less, 10 pM or less, 5 pM or less or 1 pM or less.

[0235] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody has an apparent affinity for a cynomolgus C5aR peptide comprising SEQ ID NO: 14 with a K.sub.D of 300 nM or less, such as 250 nM or less, 200 nM or less, 150 nM or less, 100 nM or less, 90 nM or less, 80 nM or less, 70 nM or less, 60 nM or less, 50 nM or less, 40 nM or less, 30 nM or less, 20 nM or less, 10 nM or 1 nM or less.

[0236] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody has an apparent affinity to a human C5aR peptide comprising SEQ ID NO: 13 with a K.sub.D of 0.3 nM or less and to a cynomolgus C5aR peptide comprising SEQ ID NO: 14 with a K.sub.D of 150 nM or less.

[0237] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody has an apparent affinity for a human C5aR peptide comprising SEQ ID NO: 13 with a K.sub.D of 0.05 nM or less and for a cynomolgus C5aR peptide comprising SEQ ID NO: 14 with a K.sub.D of 80 nM or less.

[0238] In certain embodiments, said apparent affinity is determined in IgG format. In an embodiment, said apparent affinity is determined by biolayer interferometry (BLI) as described herein in Example 5.

[0239] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR, comprises [0240] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0241] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0242] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0243] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0244] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0245] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0246] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or [0247] a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0248] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0249] a) the FIC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0250] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0251] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0252] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

Apparent Affinity (Bivalent) for Full-Length C5aR

[0253] In further embodiments, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody has an apparent affinity for human C5aR with a K.sub.D of 10 nM or less, such as 5 nM or less, 4 nM or less, 3 nM or less, 2 nM or less, 1 nM or less, 0.5 nM or less, 0.4 nM or less, 0.3 nM or less, 0.2 nM or less or 0.1 nM or less.

[0254] In embodiments, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody has an apparent affinity for cynomolgus C5aR with a K.sub.D of 10 nM or less, such as 9 nM or less, 8 nM or less, 7 nM or less, 6 nM or less, 5 nM or less, 4 nM or less, 3 nM or less, or 1 nM or less.

[0255] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody has an apparent affinity to human C5aR with a K.sub.D of 0.5 nM or less and to cynomolgus C5aR with a K.sub.D of 5 nM or less.

[0256] In an embodiment, said human C5aR comprises the amino acid sequence of SEQ ID NO: 1. In an embodiment, said human C5aR comprises the amino acid sequence of SEQ ID NO: 2. In an embodiment, said cynomolgus C5aR comprises the amino acid sequence of SEQ ID NO: 3.

[0257] In certain embodiments, said apparent affinity is determined in IgG format. In embodiments, said human C5aR or cynomolgus C5aR is expressed on cells. In embodiments, said human C5aR or cynomolgus C5aR is expressed on engineered CHO cells expressing full-length human. In embodiments, said CHO cells are Flp-In CHO cells.

[0258] In certain embodiments, said apparent affinity is determined as described herein in Example 6.

[0259] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR, comprises [0260] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0261] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34, or [0262] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34, or [0263] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34.

[0264] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0265] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0266] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or [0267] a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0268] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0269] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0270] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0271] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0272] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

Apparent EC.sub.50 for Full-Length C5aR

[0273] In further embodiments, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody binds to human C5aR with an EC.sub.50 concentration of 20 nM or less, 15 nM or less, 10 nM or less, 5 nM or less, 4 nM or less, 3 nM or less, 2 nM or less, 1 nM or less, 0.5 nM or less, 0.4 nM or less, 0.3 nM or less, 0.2 nM or less or 0.1 nM or less.

[0274] In embodiments, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody binds to cynomolgus C5aR with an EC.sub.50 concentration of 20 nM or less, 10 nM or less, 9 nM or less, 8 nM or less, 7 nM or less, 6 nM or less, 5 nM or less, 4 nM or less, 3 nM or less, or 1 nM or less.

[0275] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody binds to human C5aR and cynomolgus C5aR with an EC.sub.50 concentration of K.sub.D of 10 nM or less.

[0276] In an embodiment, said human C5aR comprises the amino acid sequence of SEQ ID NO: 1. In an embodiment, said human C5aR comprises the amino acid sequence of SEQ ID NO: 2. In an embodiment, said cynomolgus C5aR comprises the amino acid sequence of SEQ ID NO: 3.

[0277] In certain embodiments, said EC.sub.50 concentration is determined in IgG format. In embodiments, said human C5aR or cynomolgus C5aR is expressed on cells. In embodiments, said human C5aR or cynomolgus C5aR is expressed on engineered CHO cells expressing full-length human. In embodiments, said CHO cells are Flp-In CHO cells. In certain embodiments, said human C5aR or cynomolgus C5aR is expressed on neutrophils. In embodiments, said human C5aR is expressed on human neutrophils. In embodiments, said cynomolgus C5aR is expressed on cynomolgus neutrophils. In certain embodiments, said neutrophils are derived from whole-blood. In certain embodiments, said EC.sub.50 concentration is determined as described herein in Example 7.

[0278] In further embodiments, said isolated antibody or antibody fragment specific for human C5aR does substantially not bind to a C5aR related antigen selected from the group consisting of human C5L2, human ChemR23, human FPR1 and C3aR. In certain embodiments, said isolated antibody or antibody fragment specific for human C5aR does not substantially bind to a C5aR related antigen selected from the group consisting of human C5L2, human ChemR23, human FPR1 and C3aR at an IgG concentration of 300 nM. In an embodiment, said binding is determined as described in Example 7.

[0279] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR, comprises [0280] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0281] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34, or [0282] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34, or [0283] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34.

[0284] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0285] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0286] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or [0287] a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0288] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0289] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0290] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0291] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 37 or 44 and to the VL of SEQ ID NO: 38 or 45.

[0292] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

[0293] In an embodiment, the present disclosure refers to an isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment binds to human C5aR comprising SEQ ID NO: 1 and/or SEQ ID NO: 2 with an EC.sub.50 concentration of 5 nM or less, such as 4 nM or less, 3 nM or less, 2 nM or less, 1 nM or less or 0.5 nM or less.

[0294] In certain embodiments, said human C5aR is presented on virus-like-particles. In certain embodiments, said human C5aR is expressed as a fusion protein. In certain embodiments, said human C5aR is fused to a GAG protein. In embodiments, said fusion protein comprises an amino acid sequence disclosed in Table 8. In certain embodiments, said binding is determined by ELISA as described in Example 8.

[0295] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR, comprises [0296] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0297] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0298] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0299] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0300] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR [0301] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0302] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or [0303] a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0304] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR [0305] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0306] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0307] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0308] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

Functionality--C5A-Induced Activation of C5aR

[0309] In general, the isolated antibody or antibody fragment specific for human C5aR according to the present disclosure can be used to prevent or to inhibit the interaction between human C5aR and human C5a, thereby preventing, inhibiting, neutralizing or reducing the signaling pathways that are mediated by C5aR and/or modulating the biological pathways and mechanisms in which C5aR is involved.

[0310] In an embodiment, the present disclosure pertains to an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody or antibody fragment specifically interferes with C5aR-mediated signal transduction.

[0311] In a further embodiment of the present disclosure, the isolated antibody or antibody fragment specific for human C5aR specifically interferes with the interaction of C5a with C5aR expressed on cells. In yet a further embodiment, said isolated antibody or antibody fragment is capable of specifically interfering with C5a induced signal transduction mediated by C5aR.

[0312] Methods for assaying for functional activity of a C5aR specific antibody may utilize binding assays, such as the enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), fluorescence activated cell sorting (FACS) and other methods that are well known in the art (see Hampton, R. et al. (1990; Serological Methods a Laboratory Manual, APS Press, St Paul, Minn.) and Maddox, D. E. et al. (1983; J. Exp. Med. 158:1211-1216)). Alternatively, assays may test the ability of the isolated antibody or antibody fragment of the present disclosure in eliciting a biological response as a result of binding to C5aR, either in vivo or in vitro. Such assays are described in the Examples disclosed herein. Other suitable assays will be known to those of skill in the art.

[0313] In certain embodiments, the isolated antibody or antibody fragment of the present disclosure antagonizes human C5aR activity. In an embodiment, the isolated antibody or antibody fragment neutralizes human C5aR activity. In an embodiment, the isolated antibody or antibody fragment of the present disclosure inhibits human C5aR activity. In an embodiment, the isolated antibody or antibody fragment of the present disclosure inhibits human C5aR signaling. In an embodiment, said human C5aR activity or human C5aR signaling is induced by C5a. In an embodiment, said human C5aR activity or human C5aR signaling is induced by the interaction of human C5a with human C5aR. In an embodiment, said C5aR activity or C5aR signaling is induced by the binding of human C5a to human C5aR. In an embodiment, said C5aR is expressed on cells. In an embodiment, said human C5aR activity or human C5aR signaling is inhibited in vitro and/or ex vivo and/or in vivo.

C5A-Induced .beta.-Arrestin Recruitment

[0314] The ability of the isolated antibody or antibody fragment specific for human C5aR according to the present disclosure to inhibit C5a induced C5aR activation was tested in a .beta.-arrestin recruitment assay as described in Example 14 and revealed that both antibodies are even more potent antagonists of C5aR activity when compared to the prior art antibody RefMAB#1, in particular at high pathophysiological C5a concentrations.

[0315] Accordingly, in an embodiment of the present disclosure, the isolated antibody or antibody fragment specific for human C5aR inhibits the ability of C5a to induce C5aR activity. In an embodiment, said C5a induced C5aR activity is determined in a .beta.-arrestin recruitment assay as described in Example 14. In an embodiment, said C5a induced C5aR activity is determined in vitro.

[0316] In one such embodiment, the present disclosure provides an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody or antibody fragment inhibits the ability of human C5a to induce human C5aR-mediated .beta.-arrestin recruitment. In an embodiment, said isolated antibody or antibody fragment inhibits human C5a induced .beta.-arrestin recruitment. In an embodiment, said isolated antibody or antibody fragment inhibits human C5aR mediated .beta.-arrestin recruitment.

[0317] In a further embodiment of the present disclosure, the isolated antibody or antibody fragment specific for human C5aR inhibits human C5a induced human C5aR interaction with .beta.-arrestin. In one such embodiment, said human C5a induced human C5aR interaction with .beta.-arrestin and/or that human C5a induced beta-arrestin recruitment is measured using beta-galactosidase enzyme fragment complementation. In an embodiment, said human C5a induced human C5aR interaction with .beta.-arrestin and/or that human C5a induced .beta.-arrestin recruitment is determined as described in Example 14. In one such embodiment, said human C5a induced human C5aR interaction with .beta.-arrestin and/or that human C5a induced .beta.-arrestin recruitment is tested at an IgG concentration of 50 nM.

[0318] The ability of an isolated antibody or antibody fragment according to the present disclosure to inhibit human C5a induced human C5aR activity, such as to inhibit human C5a induced human C5aR interaction with .beta.-arrestin and/or human C5a induced human C5aR mediated .beta.-recruitment can be determined by generating dose-response curves for increasing concentrations of human C5a and a fixed concentration of IgG and by calculating respective EC50 concentrations.

[0319] In an embodiment, the present disclosure provides an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody or antibody fragment increases the EC.sub.50 concentration determined for human C5a in a .beta.-arrestin recruitment assay by at least 5-fold or more, such as at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 11-fold, at least 12-fold or at least 13-fold at an IgG concentration of 50 nM when compared to the EC.sub.50 concentration determined in the absence of said isolated antibody or antibody fragment.

[0320] In an embodiment, the present disclosure provides an isolated antibody or antibody fragment specific for human C5aR, wherein said isolated antibody or antibody fragment increases the EC.sub.50 concentration determined for human C5a in a .beta.-arrestin recruitment assay at an IgG concentration of 50 nM of about 5-fold, of about 6-fold, of about 7-fold, of about 8-fold, of about 9-fold, of about 10-fold, of about 11-fold, of about 12-fold or of about 13-fold when compared to the EC.sub.50 concentration determined in the absence of said isolated antibody or antibody fragment.

[0321] In an embodiment, the present disclosure provides an antibody or antibody fragment specific for human C5aR, wherein said human C5a needs to be present in an at least 5-fold or higher, such as at least 6-fold or higher, at least 7-fold or higher, at least 8-fold or higher, at least 9-fold or higher, at least 10-fold or higher, at least 11-fold or higher, at least 12-fold or higher, or at least 13-fold or higher concentration in order to induce the same human C5aR activity in a .beta.-arrestin recruitment assay at an IgG concentration of 50 nM when compared to the concentration of human C5a in the absence of said antibody or antibody fragment.

[0322] In an embodiment, said .beta.-arrestin recruitment assay is performed as described in Example 14. In an embodiment, said .beta.-arrestin recruitment assay is performed in vitro.

[0323] Alternatively, the ability of the isolated antibody or antibody fragment specific for C5aR according to the present disclosure to inhibit C5a induced C5aR mediated .beta.-arrestin recruitment can be determined by calculating the % inhibition for different human C5a concentrations.

[0324] In one such embodiment, the isolated antibody or antibody fragment specific for C5aR according to the present disclosure inhibits human C5a induced human C5aR mediated .beta.-arrestin recruitment by at least 50%, by at least 55%, by at least 60%, by at least 70%, by at least 80% or by at least 90% at an IgG concentration of 50 nM and in the presence of 1.2 nM or 11 nM human C5a compared to the level of human C5a induced human C5aR mediated .beta.-arrestin recruitment in the presence of 1.2 nM or 11 nM human C5a and absence of said antibody or antibody fragment.

[0325] In one such embodiment, the isolated antibody or antibody fragments specific for C5aR according to the present disclosure inhibits human C5a induced human C5aR mediated beta-arrestin recruitment by at least 25%, such as by at least 30%, by at least 35%, by at least 40%, by at least 45% or by at least 50% at an IgG concentration of 50 nM and in the presence of 1.2 nM or 11 nM human C5a compared to the level of human C5a induced human C5aR mediated .beta.-arrestin recruitment in the presence of 100 nM human C5a and absence of said antibody or antibody fragment.

[0326] In an embodiment, said .beta.-arrestin recruitment assay is performed as described in Example 14. In an embodiment, said .beta.-arrestin recruitment assay is performed in vitro.

[0327] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0328] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0329] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0330] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0331] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region comprising the amino acid sequence of SEQ ID NO: 34.

[0332] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR for comprises [0333] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0334] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or [0335] a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0336] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR for comprises [0337] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0338] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0339] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0340] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

C5A-Induced Upregulation of CD11b Expression

[0341] C5a, as a potent activator of human neutrophils and monocytes, induces up-regulation of the cell surface antigen CD11b in such cells. Thus, the ability of an isolated antibody or antibody fragment specific for human C5aR according to the present disclosure to inhibit C5a induced activation of granulocytes and/or monocytes can be assessed by determine CD11b expression levels in such cells.

[0342] The ability of the isolated antibody or antibody fragments according to the present disclosure to inhibit CD11b expression in granulocytes and/or monocytes can be determined by generating dose-response curves for increasing concentrations of IgG and fixed concentration of human C5a and calculating the respective IC.sub.50 concentrations. Alternatively, the ability of an isolated antibody or antibody fragments according to the present disclosure to inhibit CD11b expression in granulocytes and/or monocytes C5a can be determined by calculating the % inhibition of CD11b expression for different IgG concentrations.

[0343] In one such embodiment, the isolated antibody or antibody fragment specific for human C5aR according to the present disclosure inhibits human C5a induced CD11b expression in human granuloyctes and/or human monocytes with an IC.sub.50 concentration of 30 nM or less, 25 nM or less, 20 nM or less, 15 nM or less, 10 nM or less or 5 nM or less, in the presence of 15 nM human C5a.

[0344] In another embodiment, the isolated antibody or antibody fragment specific for human C5aR according to the present disclosure inhibits human C5a induced CD11b expression in human granuloyctes and/or human monocytes by at least 70%, by at least 75%, by at 80%, by at least 85% or by at least 90% in the presence of 15 nM human C5a and an IgG concentration of 600 nM compared to the level of CD11b expression in the presence of 15 nM human C5a and absence of said antibody or antibody fragment.

[0345] In a further embodiment, the isolated antibody or antibody fragment specific for human C5aR according to the present disclosure inhibits human C5a induced CD11b expression in human granuloyctes with an IC.sub.50 concentration of 150 nM or less, 125 nM or less, 100 nM or less, 90 nM or less, 80 nM or less, 70 nM or less, 60 nM or less, 50 nM or less, or 40 nM or less in the presence of 150 nM human C5a.

[0346] In an embodiment, the isolated antibody or antibody fragment specific for human C5aR according to the present disclosure inhibits human C5a induced CD11b expression in human granulocytes with an IC.sub.50 concentration of 42 nM in the presence of 150 nM human C5a.

[0347] In a further embodiment, the isolated antibody or antibody fragment specific for human C5aR according to the present disclosure inhibits human C5a induced CD11b expression in human granuloyctes by at least by at least 65%, at least 70%, at least 75%, at least 80% or at least 90% in the presence of 150 nM human C5a and an IgG concentration of 600 nM compared to the level of CD11b expression in the presence of 150 nM human C5a and absence of said antibody or antibody fragment.

[0348] In an embodiment, the isolated antibody or antibody fragments specific for human C5aR according to the present disclosure inhibits C5a induced CD11b expression in human granuloyctes by at least 45%, at least 50%, at least 55%, at least 60% or at least 65% in the presence of 150 nM human C5a and an IgG concentration of 100 nM compared to the level of CD11b expression in the presence of 150 nM human C5a and absence of said antibody or antibody fragment.

[0349] In an embodiment, said determination of CD11b expression is performed as described in Example 15. In an embodiment, said determination of CD11b expression in performed in vitro and/or ex vivo.

[0350] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0351] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0352] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0353] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0354] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0355] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR for comprises [0356] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0357] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or [0358] a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0359] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR for comprises [0360] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0361] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0362] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0363] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

[0364] Furthermore, the inhibitory activity of the isolated antibody or antibody fragment according to the present disclosure on human C5a induced CD11b expression was further analyzed over a prolonged period of time meaning that the antibodies were pre-incubated with granulocytes and/or monocytes present in whole-blood over time of varying length (e.g. 300 minutes vs. 20 minutes) before human C5a was added. Surprisingly, the experiment revealed that the isolated antibodies according to the present disclosure are even more potent antagonists of C5aR activity over a longer period of time, e.g. a prolonged period of incubation time, in particular when compared to the prior art antibody RefMAB#1.

[0365] Accordingly, the ability of the isolated antibody or antibody fragment according to the present disclosure to inhibit human C5a induced CD11b expression in granulocytes and/or monocytes over a prolonged period of time can be determined by generating dose-response curves for increasing concentrations of IgG and a fixed concentration of human C5a and by calculating respective EC.sub.50 values after incubating said isolated antibodies with said granulocytes and/or monocytes for different incubation times.

[0366] As shown in FIGS. 6A and C, the isolated antibodies or antibody fragments of the present disclosure exhibited a clear shift of the determined dose-response curve to lower IgG concentrations determined after 300 minutes of incubation when compared to the dose-response curve determined after 20 minutes of incubation. Interestingly, RefMAB#1 revealed no increased potency over time (see FIGS. 6B and D).

[0367] Thus, in an embodiment, the isolated antibody or antibody fragment according to the present disclosure is more potent in inhibiting C5a induced CD11b expression in human granuloyctes and/or human monocytes after a prolonged period of incubation time with said cells.

[0368] In one such embodiment, the isolated antibody or antibody fragment according to the present disclosure inhibits human C5a induced CD11b expression in human granuloyctes and/or human monocytes with an at least 2-fold, at least 4-fold, at least 5-fold or at least 6-fold lower IC.sub.50 concentration determined after a prolonged period of incubation time of 50 minutes, of 100 minutes, of 150 minutes, of 200 minutes, of 250 minutes, or of 300 minutes when compared to the IC.sub.50 concentration determined after a period of incubation time of 20 minutes.

[0369] In an embodiment, the isolated antibody or antibody fragment according to the present disclosure inhibits human C5a induced CD11b expression in human granuloyctes and/or human monocytes with an about 5-fold lower IC.sub.50 concentration determined after a prolonged period of incubation time of 300 minutes when compared to the IC.sub.50 concentration determined after a period of incubation time of 20 minutes.

[0370] In an embodiment, the isolated antibody or antibody fragment according to the present disclosure inhibits human C5a induced CD11b expression in human granuloyctes and/or human monocytes with an at least 3-fold, at least 4-fold, at least 5-fold, at least 10-fold, at least 15-fold or at least 19-fold lower IC.sub.50 concentration determined after a prolonged period of incubation time of 300 minutes when compared to the corresponding IC.sub.50 concentration of RefMAB#1.

[0371] In an embodiment, the isolated antibody or antibody fragment according to the present disclosure inhibits human C5a induced CD11b expression in human granuloyctes and/or human monocytes with an IC.sub.50 concentration of 3 nM or less, 2.5 nM or less, 2 nM or less, 1.5 nM or less or 1 nM or less, after a prolonged period of incubation time of 300 minutes with said cells.

[0372] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0373] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0374] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0375] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0376] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0377] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0378] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0379] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or [0380] a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0381] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0382] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0383] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0384] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0385] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

C5A-Induced Migration of Neutrophils

[0386] In further assays, the ability of the isolated antibody or antibody fragment specific for human C5aR according to the present disclosure to inhibit human C5a induced migration of human neutrophils was evaluated and revealed that MAB#1 efficiently inhibited C5 induced neutrophil migration in vitro.

[0387] Thus, in an embodiment of the present disclosure, said isolated antibody or antibody fragment specific for human C5aR of the present disclosure inhibits human C5a induced migration of human neutrophils in vitro.

[0388] In a further embodiment, said isolated antibody or antibody fragment inhibits human C5a induced migration of human neutrophils by at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75 or at least 80% compared to the level of migration in the presence of 10 nM human C5a and absence of said isolated antibody or antibody fragment in vitro.

[0389] In an embodiment, said migration of human neutrophils is determined after 15 minutes, after 25 minutes and/or after 35 minutes. In certain embodiments, said isolated antibody or antibody fragment according to the present disclosure is tested at an IgG concentration of 100 nM and/or 600 nM.

[0390] In an embodiment, said isolated antibody or antibody fragment of the present disclosure inhibits human C5a induced migration of human neutrophils by at least 25% after 35 minutes and at an IgG concentration of 100 nM compared to the level of migration after 35 minutes in the presence of 10 nM human C5a and absence of said antibody or antibody fragment.

[0391] In an embodiment, said isolated antibody or antibody fragment of the present disclosure inhibits human C5a induced migration of human neutrophils by at least 60% after 25 minutes and at an IgG concentration of 100 nM and/or 600 nM compared to the level of migration after 25 minutes in the presence of 10 nM human C5a and absence of said antibody or antibody fragment.

[0392] In an embodiment, said isolated antibody or antibody fragment of the present disclosure inhibits human C5a induced migration of human neutrophils by at least 40% after 35 minutes at an IgG concentration of 600 nM compared to the level of migration after 35 minutes in the presence of 10 nM human C5a and absence of said antibody.

[0393] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0394] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0395] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0396] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0397] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0398] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0399] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0400] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or [0401] a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0402] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0403] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0404] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0405] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0406] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

Effector Function

[0407] The Fc region of an immunoglobulin generally confers to the favorable pharmacokinetic properties of antibodies, such as prolonged half-life in serum and to the ability to induce effector function via binding to Fc receptors expressed on cells. On the other hand, binding to Fc receptors might also results in an undesirable activation of certain cell surface receptors leading to unwanted cytokine release and severe side effects upon systemic administration.

[0408] Accordingly, for certain therapeutic situations, it is desirable to reduce or abolish the normal binding of the wild-type Fc region of an antibody, such as of an wild-type IgG Fc region to one or more or all of Fc receptors and/or binding to a complement component, such as C1q in order to reduce or abolish the ability of the antibody to induce effector function. For instance, it may be desirable to reduce or abolish the binding of the Fc region of an antibody to one or more or all of the Fc.gamma. receptors, such as: Fc.gamma.RI, Fc.gamma.RIIa, Fc.gamma.RIIb, Fc.gamma.RIIIa.

[0409] Effector function can include, but is not limited to, one or more of the following: complement dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), cytokine secretion, immune complex-mediated antigen uptake by antigen-presenting cells, binding to NK cells, binding to macrophages, binding to monocytes, binding to polymorphonuclear cells, direct signaling inducing apoptosis, crosslinking of target-bound antibodies, dendritic cell maturation, or T cell priming.

[0410] A reduced or abolished binding of an Fc region to an Fc receptor and/or to C1q is typically achieved by mutating a wild-type Fc region, such as of an IgG1 Fc region, more particular a human IgG1 Fc region, resulting in a variant or engineered Fc region of said wild-type Fc region, e.g. a variant human IgG1 Fc region. Substitutions that result in reduced binding can be useful. For reducing or abolishing the binding properties of an Fc region to an Fc receptor, non-conservative amino acid substitutions, i.e. replacing one amino acid with another amino acid having different structural and/or chemical properties, are preferred.

[0411] Accordingly, in an embodiment, the isolated antibody or antibody fragment specific for human C5aR according to the present disclosure comprises a variant Fc region having a reduced or abolished binding to an Fc receptor and/or to C1q when compared to the wild-type Fc region. In one such embodiment, the isolated antibody or antibody fragment according to the present disclosure comprises a variant Fc region that reduces or abolishes the ability of the antibody to induce effector function. In a further embodiment, the isolated antibody or antibody fragment according to the present disclosure does not substantially induce effector function.

[0412] In certain embodiments, the effector function is one or more selected from the group consisting of CDC, ADCC and ADCP. In an embodiment, the effector function is ADCC. In an embodiment, the effector function is CDC. In an embodiment, the effector function is ADCP. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure does not substantially induce ADCC and/or CDC and/or ADCP. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure does not induce ADCC or ADCP in vitro.

[0413] In an embodiment, the variant Fc region of the isolated antibody or antibody fragment according to the present disclosure comprises one or more amino acid substitutions that reduce or abolish the binding of the variant Fc region to one or more Fc receptors and/or to C1q when compared to the wild-type Fc region. In an embodiment, the variant Fc region of the isolated antibody or antibody fragment according to the present disclosure comprises one or more amino acid substitutions that reduce or abolish the ability of the antibody to induce effector function when compared to the wild-type Fc region.

[0414] In a particular embodiment, the one or more amino acid substitutions may reduce the binding affinity of the variant Fc region for one or more Fc receptors and/or to C1q by at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold or even at least 50-fold when compared to the wild-type Fc region. In alternative embodiments, the one or more amino acid substitutions may reduce the ability of the isolated antibody or antibody fragment according to the present disclosure to induce effector function by at least 2-fold, at least 5-fold, at least 10-fold, at least 20-fold or even at least 50-fold when compared to the wild-type Fc region.

[0415] In an embodiment, the variant Fc region of the isolated antibody or antibody fragment according to the present disclosure does not substantially bind to one or more Fc receptors and/or C1q. In an embodiment, the variant Fc region of the antibody according to the present disclosure does substantially abolish the ability of said antibody to induce effector function. In an embodiment, the antibody or antibody fragment according to the present disclosure does not substantially induce effector function. In an embodiment, said effect function is ADCC and/or ADCP and/or CDC. In an embodiment, the antibody or antibody fragment according to the present disclosure does not substantially induce effector function meaning that the level of induced effector function is not significantly above the background as measured in the absence of said antibody.

[0416] In an embodiment, the Fc receptor is a human Fc receptor. In an embodiment, the Fc receptor is an Fc.gamma. receptor. In an embodiment, the Fc receptor is a human Fc.gamma.RIIIa, Fc.gamma.RI, Fc.gamma.RIIa and/or Fc.gamma.RIIb.

[0417] In an embodiment, the effector function is one or more selected from the group of CDC, ADCC and ADCP. In a particular embodiment, the effector function is ADCC, CDC and ADCP. In a more particular embodiment, the effector function is ADCC and ADCP.

[0418] In an embodiment, the wild-type Fc region is an IgG1 Fc region. In an embodiment, the wild-type Fc region is a human IgG1 Fc region. In an embodiment, the wild-type Fc region is a human IgG1 Fc region. In an embodiment, the wild-type Fc region is a human IgG1 Fc region comprising the amino acid sequence of:

TABLE-US-00011 (SEQ ID NO: 11) PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGK.

[0419] In an embodiment, the isolated antibody or antibody fragment according to the present disclosure comprises a variant human IgG1 Fc region, which comprises one or more amino acid substitutions compared to the wild-type human IgG1 Fc region. In an embodiment, that one or more amino acid substitutions reduce or abolish the binding of the variant Fc region to an Fc receptor and/or to C1q and/or reduces the ability of said antibody to induce effector function when compared to the wild-type Fc region.

[0420] In an embodiment, the variant human IgG1 Fc region of the antibody or antibody fragment according to the present disclosure comprises an amino acid substitution at one or more positions selected from the group of 234, 235, 237, 330 and 331 with numbering according EU index compared to the wild type IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 11.

[0421] In an embodiment, said variant human IgG1 Fc region comprises an amino acid substitution at one or more positions selected from the group of L234, L235 and G237 with numbering according EU index compared to the wild-type IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 11.

[0422] In an embodiment, the variant human IgG1 Fc region comprises the amino acid substitutions at one or more positions selected from the group of L234A, L235E G237A with numbering according EU index compared to the wild-type IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 11.

[0423] In an embodiment, the variant human IgG1 Fc region comprises the amino acid substitutions L234A and L235E with numbering according EU index compared to the wild-type IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 11.

[0424] In an embodiment, the variant human IgG1 Fc region comprises the amino acid substitutions L234A, L235E and G237A with numbering according EU index compared to the wild-type IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 11.

[0425] In an embodiment, the variant human IgG1 Fc region comprises an amino acid substitution at one or more position selected from the group of A330 and P331 with numbering according EU index compared to the wild-type IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 11.

[0426] In an embodiment, the variant human IgG1 Fc region comprises the amino acid substitutions at one or more positions selected from the group of A330S and P331S with numbering according EU index compared to the wild-type IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 11.

[0427] In an embodiment, the variant human IgG1 Fc region comprises the amino acid substitutions A330S and P331S with numbering according EU index compared to the wild-type IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 11.

[0428] In an embodiment, the variant human IgG1 Fc region comprises the amino acid substitutions L234A, L235E, G237A, A330S and P331S with numbering according EU index compared to the wild type IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 11.

[0429] In an embodiment, the antibody or antibody fragment according to the present disclosure comprises a variant human IgG1 Fc region, which comprises one or more amino acid substitutions compared to the wild-type human IgG1 Fc region comprising the sequence of SEQ ID NO: 11, that reduce or abolish the binding affinity of the variant Fc region to one or more Fc receptors and/or to C1q and/or reduces the ability of said antibody to induce effector function, wherein the one or more amino acid substitutions are L234A, L235E, G237A, A330S and P331S with numbering according EU index.

[0430] In an embodiment, the isolated antibody or antibody fragment according to the present disclosure is of the human IgG1 class. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure is of a variant human IgG1 class. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure does not substantially induce effector function in vitro. In an embodiment, the variant human IgG1 Fc region does not substantially bind to one or more Fc receptors and/or C1q. In one such embodiment, the isolated antibody or antibody fragment according to the present disclosure comprises the amino acid substitution selected from the group of: L234A, L235E, G237A, A330S and P331S, with numbering according to EU index compared to the wild type IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 11.

[0431] In an embodiment, the isolated antibody or antibody fragment according to the present disclosure is of the human IgG1 class comprising the amino acid substitution L234A, L235E, G237A, A330S and P331S, with numbering according to EU index.

[0432] In an embodiment, the isolated antibody or antibody fragment according to the present disclosure comprises the amino acid substitution L234A, L235E, G237A, A330S and P331S, with numbering according to EU index.

[0433] In an embodiment, the isolated antibody or antibody fragment according to the present disclosure comprises the amino acid substitution L234A, L235E, G237A, A330S and P331S, with numbering according to EU index compared to the wild type IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 11.

[0434] In one such embodiment, the isolated antibody or antibody fragment according to the present disclosure comprises the amino acid substitution L234A, L235E, G237A, A330S and P331S, with numbering according to EU index and does not substantially induce effector function in vitro. In an embodiment, the effector function is one or more selected from the group of CDC, ADCC and ADCP.

[0435] In one such embodiment, the isolated antibody or antibody fragment according to the present disclosure comprises the amino acid substitution L234A, L235E, G237A, A330S and P331S, with numbering according to EU index compared to the wild-type human IgG1 Fc region comprising the amino acid sequence of SEQ ID NO: 11 and does not substantially bind to one or more Fc receptors and/or C1q in vitro.

[0436] In an embodiment, the isolated antibody or antibody fragment according to the preset disclosure is of the human IgG1 class. In an embodiment, the isolated antibody or antibody fragment to the preset disclosure does not substantially induce effector function in vitro. In an embodiment, the isolated antibody or antibody fragment according to the present disclosure comprises one or more amino acid substitution selected from the group of: L234A, L235E, G237A, A330S and P331S, with numbering according to EU index.

[0437] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0438] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0439] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0440] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0441] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0442] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR for comprises [0443] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0444] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or [0445] a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0446] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR for comprises [0447] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0448] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0449] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0450] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

[0451] Binding of an antibody to Fc receptors via its Fc region can be easily determined e.g. by ELISA, or by Surface Plasmon Resonance (SPR) using standard instrumentation such as a Biacore.RTM. instrument (GE Healthcare), and Fc receptors may be obtained by recombinant expression. Alternatively, the binding affinity of Fc regions may be evaluated using cell lines known to express particular Fc receptors, such as NK cells expressing Fc.gamma.IIIa receptor. Effector function of an antibody can be measured by methods known in the art. Suitable in vitro assays to assess ADCC activity of a molecule of interest are for instance described in WO2012130831. Useful effector cells for such assays include peripheral blood mononuclear cells (PBMC) and Natural Killer (NK) cells. Alternatively, or additionally, ADCC activity of the molecule of interest may be assessed in vivo, e.g. in an animal model such as that disclosed in Clynes et al., Proc Natl Acad Sci USA 95, 652-656 (1998). To assess complement activation, a CDC assay may be performed (see, for example, Gazzano-Santoro et al., J Immunol Methods 202, 163 (1996); Cragg et al., Blood 101, 1045-1052 (2003); and Cragg and Glennie, Blood 103, 2738-2743 (2004)). C1q binding assays (such as ELISA) may be carried out to determine whether an antibody is able to bind C1q and hence has CDC activity (see, for example WO 2006/029879). In vitro methods to asses binding to Fc receptors or to asses immune effector function are described herein in Examples 10-13.

Fusion Proteins

[0452] The isolated antibody or antibody fragment according to the present disclosure may or may not be fused to one or more other amino acid residues, polypeptides or moieties. Such a fusion protein may be prepared in any suitable manner, including genetically or chemically approaches. Said linked moieties may contain secretory or leader sequences, sequences that aid detection, expression, separation or purification, or sequences that confer to increased protein stability, for example, during recombinant production. Non-limiting examples of potential moieties include beta-galactosidase, glutathione-S-transferase, luciferase, a T7 polymerase fragment, a secretion signal peptide, an antibody or antibody fragment, a toxin, a reporter enzyme, a moiety being capable of binding a metal ion like a poly-histidine tag, a tag suitable for detection and/or purification, a homo- or hetero-association domain, a moiety which increases solubility of a protein, or a moiety which comprises an enzymatic cleavage site.

[0453] Accordingly, the isolated antibody or antibody fragment according to the present disclosure may optionally contain one or more moieties for binding to other targets or target proteins of interest. It should be clear that such further moieties may or may not provide further functionality to the antibody and may or may not modify the properties of the isolated antibody or antibody fragment according to the present disclosure. Diagnostic use

[0454] In an embodiment, the present disclosure provides the use of an isolated antibody or antibody fragment specific for human C5aR according to the present disclosure for the diagnosis of a disease. In an embodiment, the present disclosure provides the use of an antibody or antibody fragment according to the present disclosure for the detection of C5aR, in particular human C5aR and/or cynomolgus C5aR. In an embodiment, the present disclosure provides a method for detecting C5aR in a subject or a sample, comprising the step of contacting said subject or sample with an isolated antibody or antibody fragment specific for human C5aR of the present disclosure. In an embodiment, the present disclosure provides a method for diagnosing a disease in a subject, comprising the step of contacting said subject or sample with an isolated antibody or antibody fragment according to the present disclosure.

[0455] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0456] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0457] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0458] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0459] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0460] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR for comprises [0461] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0462] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or [0463] a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0464] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR for comprises [0465] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0466] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0467] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0468] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

Therapeutic Methods

[0469] The isolated antibody or antibody fragment according to the present disclosure may be used in therapeutic methods. The antibody or antibody fragment according to the present disclosure may be used for the treatment of a disease, such as cancer, an autoimmune disease or inflammatory disease.

[0470] In an embodiment, the present disclosure provides a method for the treatment of a disease.

[0471] In an embodiment, the present disclosure provides an isolated antibody or antibody fragment according to the present disclosure for the treatment of a disease. In an embodiment, the present disclosure provides an isolated antibody or antibody fragment according to the present disclosure for use in the treatment of a disease. In an embodiment, the present disclosure provides an isolated antibody or antibody fragment according to the present disclosure for use in the treatment of a disease in a subject in need thereof.

[0472] In an embodiment, the present disclosure provides the use of an isolated antibody or antibody fragment according to the present disclosure for the manufacture of a medicament. In an embodiment, the present disclosure provides an isolated antibody or antibody fragment according to the present disclosure for use as a medicament. In an embodiment, the present disclosure provides an isolated antibody or antibody fragment according to the present disclosure for use in medicine. In an embodiment, the present disclosure provides an isolated antibody or antibody fragment according to the present disclosure for use as a medicament for the treatment of a subject in need thereof.

[0473] In an embodiment, the disease is associated with the undesired presence of C5aR, in particular human C5aR. In an embodiment, the disease is associated with the undesired presence of C5a, in particular human C5a.

[0474] In an embodiment, the disease to be treated is a proliferative disease. In a particular embodiment, the disease is cancer. Non-limiting examples of cancers include bladder cancer, brain cancer, head and neck cancer, pancreatic cancer, lung cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, endometrial cancer, esophageal cancer, colon cancer, colorectal cancer, rectal cancer, gastric cancer, prostate cancer, blood cancer, sarcoma, skin cancer, squamous cell carcinoma, bone cancer, melanoma, renal cell carcinoma, and kidney cancer.

[0475] In an embodiment, the disease to be treated is an autoimmune or inflammatory disease. Non-limiting examples an autoimmune or inflammatory disease include rheumatoid arthritis (RA), psoriasis, psoriatic arthritis, systemic lupus erythematosus (SLE), lupus nephritis, type I diabetes, Grave's disease, Inflammatory bowel disease (IBD), Crohn's disease (CD), ulcerative colitis (UC), irritable bowel syndrome, multiple sclerosis (MS), autoimmune myocarditis, Kawasaki disease, coronary artery disease, chronic obstructive pulmonary disease (COPD), interstitial lung disease, autoimmune thyroiditis, scleroderma, systemic sclerosis, osteoarthritis, atoptic dermatitis, vitiligo, graft vs. host disease, Sjogren's syndrome, autoimmune nephritis, Goodpasture's syndrome, chronic inflammatory demyelinating polyneuropathy, ANCA-associated vasculitis, uveitis, scleroderma, bullous pemphigoid, Alzheimer's Disease, amyotrophic lateral sclerosis, Huntington's Chorea, cystic fibrosis, gout, age-related macular degeneration, allergy, asthma, antiphospholipid syndrome (APS), atherosclerosis, C3 glomerulopathy and IgA nephropathy, ischemia/reperfusion injury, peritonitis, sepsis and other autoimmune diseases that are a result of either acute or chronic inflammation.

[0476] In an embodiment, the present disclosure provides an isolated antibody or antibody fragment specific for human C5aR according to the present disclosure for use in a method of treating a subject having a disease comprising administering to the subject a therapeutically effective amount of an antibody or antibody fragment according to the present disclosure.

[0477] In an embodiment, the method further comprises administering to the subject a therapeutically effective amount of at least one additional therapeutic agent. The subject in need of treatment is typically a mammal, more specifically a human. For use in therapeutic methods, an isolated antibody or antibody fragment according to the present disclosure would be formulated, dosed, and administered in a way consistent with good medical practice.

[0478] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0479] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0480] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0481] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0482] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 of SEQ ID NO: 34.

[0483] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0484] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0485] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or [0486] a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0487] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR comprises [0488] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0489] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0490] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the HC of SEQ ID NO: 37 or 44 and to the LC of SEQ ID NO: 38 or 45.

[0491] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

Pharmaceutical Compositions

[0492] In an embodiment, the present disclosure provides a pharmaceutical composition comprising an isolated antibody or antibody fragment according to the present disclosure and a pharmaceutically acceptable carrier or excipient.

[0493] The pharmaceutical compositions may further comprise at least one other pharmaceutically active compound. The pharmaceutical composition according to the present disclosure can be used in the diagnosis, prevention and/or treatment of diseases associated with the undesired presence of C5aR, in particular human C5aR. In particular, the present disclosure provides a pharmaceutical compositions comprising an antibody or antibody fragment according to the present disclosure that is suitable for prophylactic, therapeutic and/or diagnostic use in a mammal, more particular in a human.

[0494] In general, an antibody or antibody fragment according to the present disclosure may be formulated as a pharmaceutical composition comprising at least one antibody or antibody fragment according to the present disclosure and at least one pharmaceutically acceptable carrier or excipient, and optionally one or more further pharmaceutically active compounds. Such a formulation may be suitable for oral, parenteral, topical administration or for administration by inhalation. Accordingly, a pharmaceutical composition comprising at least one antibody or antibody fragment according to the present disclosure may be administered parenterally, such as intravenously, or intramuscularly, or subcutaneously. Alternatively, an antibody of the invention may be administered via a non-parenteral route, such as per-orally or topically. In a preferred embodiment, a pharmaceutical composition comprising an antibody or antibody fragment according to the present disclosure is administered intravenously or subcutaneously.

[0495] In particular, an antibody or antibody fragment according to the present disclosure may be used in combination with one or more pharmaceutically active compounds that are or can be used for the prevention and/or treatment of the diseases in which a target antigen of interest is involved, as a result of which a synergistic effect may or may not be obtained. Examples of such compounds, as well as routes, methods and pharmaceutical formulations or compositions for administering them will be clear to the clinician.

[0496] In an embodiment, the present disclosure provides a pharmaceutical composition comprising an antibody or antibody fragment according to the present disclosure for use in the prevention and/or treatment of a disease associated with the undesired presence of C5aR. In an embodiment, the present disclosure provides a pharmaceutical composition comprising an antibody or antibody fragment according to the present disclosure for the use as a medicament. In an embodiment, the present disclosure provides a pharmaceutical composition comprising an antibody or antibody fragment according to the present disclosure for use in the prevention and/or treatment of an autoimmune disease and/or inflammatory disease and/or cancer.

[0497] In an embodiment, the present disclosure provides a method for the treatment of an autoimmune disease and/or inflammatory disease and/or cancer in a subject in need thereof using a pharmaceutical composition comprising an antibody or antibody fragment according to the present disclosure.

[0498] Further provided is a method of producing an antibody or antibody fragment according to the present disclosure in a form suitable for administration in vivo, the method comprising (a) obtaining an antibody or antibody fragment by a method according to the present disclosure, and (b) formulating said antibody or antibody fragment with at least one pharmaceutically acceptable carrier or excipient, whereby a preparation of antibody or antibody fragment is formulated for administration in vivo. Pharmaceutical compositions according to the present disclosure comprise a therapeutically effective amount of one or more antibodies or antibody fragments according to the present disclosure dissolved in a pharmaceutically acceptable carrier or excipient.

[0499] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR according to the present disclosure comprises [0500] a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0501] b) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 31, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0502] c) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or [0503] d) the HCDR1 region of SEQ ID NO: 30, the HCDR2 region of SEQ ID NO: 41, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

[0504] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR for comprises [0505] a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO: 36 or [0506] b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO: 43 or [0507] a VH and a VL that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 35 or 42 and to the VL of SEQ ID NO: 36 or 43.

[0508] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR for comprises [0509] a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO: 38 or [0510] b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO: 45 or [0511] a HC and a LC that has at least at least 80%, at least 85%, at least 90% or at least 95% identity to the VH of SEQ ID NO: 37 or 44 and to the VL of SEQ ID NO: 38 or 45.

[0512] In an embodiment, said isolated antibody or antibody fragment specific for human C5aR is a monoclonal antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a recombinant antibody or antibody fragment. In an embodiment, said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

Antibody Sequences

TABLE-US-00012 [0513] TABLE 1 Antibody sequences of MAB#1 MAB#1 SEQ ID NO: [aa] / DNA MAB#1 Protein HCDR1 (Kabat) SEQ ID NO: 27 SYAMH HCDR2 (Kabat) SEQ ID NO: 28 RIKSKAQGGTTDYAAHVKG HCDR3 (Kabat) SEQ ID NO: 29 VSFSTFDV HCDR1 (Chothia) SEQ ID NO: 30 GFTFSSY HCDR2 (Chothia) SEQ ID NO: 31 KSKAQGGT HCDR3 (Chothia) SEQ ID NO: 29 VSFSTFDV LCDR1 (Kabat) SEQ ID NO: 32 SGSSSNIGSYYVS LCDR2 (Kabat) SEQ ID NO: 33 RNNQRPS LCDR3 (Kabat) SEQ ID NO: 34 DSWDHSSMNV LCDR1 (Chothia) SEQ ID NO: 32 SGSSSNIGSYYVS LCDR2 (Chothia) SEQ ID NO: 33 RNNQRPS LCDR3 (Chothia) SEQ ID NO: 34 DSWDHSSMNV VH SEQ ID NO: 35 EVQLVESGGGLVKPGGSLRLSCAASGFTFSSY AMHWVRQAPGKGLEWVGRIKSKAQGGTTDY AAHVKGRFTISRDDSKNTLYLQMNSLKTEDTA VYYCARVSFSTFDVWGQGTLVTVSS VL SEQ ID NO: 36 QSVLTQPPSVSGAPGQRVTISCSGSSSNIGSY YVSWYQQLPGTAPKVLIYRNNQRPSGVPDRF SGSKSGTSASLAITGLQAEDEADYYCDSWDH SSMNVFGGGTKLTVLGQ Heavy chain SEQ ID NO: 37 EVQLVESGGGLVKPGGSLRLSCAASGFTFSSY IgG1_AEASS AMHWVRQAPGKGLEWVGRIKSKAQGGTTDY AAHVKGRFTISRDDSKNTLYLQMNSLKTEDTA VYYCARVSFSTFDVWGQGTLVTVSSASTKGP SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVT VPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS CDKTHTCPPCPAPEAEGAPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPSSIEKTISKAKGQPREP QVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK Light chain SEQ ID NO: 38 QSVLTQPPSVSGAPGQRVTISCSGSSSNIGSY YVSWYQQLPGTAPKVLIYRNNQRPSGVPDRF SGSKSGTSASLAITGLQAEDEADYYCDSWDH SSMNVFGGGTKLTVLGQPKAAPSVTLFPPSSE ELQANKATLVCLISDFYPGAVTVAWKADSSPV KAGVETTTPSKQSNNKYAASSYLSLTPEQWKS HRSYSCQVTHEGSTVEKTVAPTECS MAB#1 DNA HCDR1 (Kabat) SEQ ID NO: 46 AGCTATGCGATGCAC HCDR2 (Kabat) SEQ ID NO: 47 CGTATCAAATCCAAAGCCCAGGGCGGTACG ACCGACTACGCGGCGCACGTGAAAGGC HCDR3 (Kabat) SEQ ID NO: 48 GTTTCTTTCTCCACTTTCGATGTT HCDR1 (Chothia) SEQ ID NO: 49 GGATTTACCTTCAGCAGCTAT HCDR2 (Chothia) SEQ ID NO: 50 AAATCCAAAGCCCAGGGCGGTACG HCDR3 (Chothia) SEQ ID NO: 48 GTTTCTTTCTCCACTTTCGATGTT LCDR1 (Kabat) SEQ ID NO: 51 AGCGGCAGCTCCTCCAATATTGGTAGCTATT ACGTGAGC LCDR2 (Kabat) SEQ ID NO: 52 CGTAATAATCAACGTCCTAGC LCDR3 (Kabat) SEQ ID NO: 53 GACAGCTGGGATCACAGCTCCATGAATGTT LCDR1 (Chothia) SEQ ID NO: 51 AGCGGCAGCTCCTCCAATATTGGTAGCTATT ACGTGAGC LCDR2 (Chothia) SEQ ID NO: 52 CGTAATAATCAACGTCCTAGC LCDR3 (Chothia) SEQ ID NO: 53 GACAGCTGGGATCACAGCTCCATGAATGTT VH SEQ ID NO: 54 GAGGTGCAATTGGTGGAAAGCGGCGGTGGC CTGGTGAAACCAGGCGGCAGCCTGCGCCTG AGCTGCGCCGCCTCCGGATTTACCTTCAGC AGCTATGCGATGCACTGGGTGCGCCAGGCC CCGGGCAAAGGTCTCGAATGGGTGGGTCGT ATCAAATCCAAAGCCCAGGGCGGTACGACC GACTACGCGGCGCACGTGAAAGGCCGCTTT ACCATTAGCCGCGATGATTCGAAAAACACCC TGTATCTGCAAATGAACAGCCTGAAAACCGA AGATACGGCCGTGTATTATTGCGCGCGTGTT TCTTTCTCCACTTTCGATGTTTGGGGCCAAG GCACCCTGGTGACTGTCTCGAGC VL SEQ ID NO: 55 CAGAGCGTGCTGACCCAGCCTCCTAGCGTG AGCGGTGCACCGGGCCAGCGCGTGACCATT AGCTGTAGCGGCAGCTCCTCCAATATTGGTA GCTATTACGTGAGCTGGTATCAGCAGCTGC CGGGCACGGCGCCGAAAGTTCTGATCTATC GTAATAATCAACGTCCTAGCGGCGTGCCGG ATCGCTTTAGCGGATCCAAAAGCGGCACCA GCGCCAGCCTGGCGATTACCGGCCTGCAAG CAGAAGATGAAGCGGATTATTACTGCGACAG CTGGGATCACAGCTCCATGAATGTTTTTGGC GGCGGTACCAAGCTGACCGTGCTGGGCCAG Heavy chain (IgG1) SEQ ID NO: 56 GAGGTGCAATTGGTGGAAAGCGGCGGTGGC IgG1 AEASS CTGGTGAAACCAGGCGGCAGCCTGCGCCTG AGCTGCGCCGCCTCCGGATTTACCTTCAGC AGCTATGCGATGCACTGGGTGCGCCAGGCC CCGGGCAAAGGTCTCGAATGGGTGGGTCGT ATCAAATCCAAAGCCCAGGGCGGTACGACC GACTACGCGGCGCACGTGAAAGGCCGCTTT ACCATTAGCCGCGATGATTCGAAAAACACCC TGTATCTGCAAATGAACAGCCTGAAAACCGA AGATACGGCCGTGTATTATTGCGCGCGTGTT TCTTTCTCCACTTTCGATGTTTGGGGCCAAG GCACCCTGGTGACTGTCTCGAGCGCGTCGA CCAAAGGCCCCAGCGTGTTCCCTCTGGCCC CCAGCAGCAAGAGCACCTCTGGCGGAACAG CCGCCCTGGGCTGCCTGGTCAAGGACTACT TCCCCGAGCCCGTGACCGTGTCCTGGAACT CTGGCGCCCTGACCAGCGGCGTGCACACCT TTCCAGCCGTGCTCCAGAGCAGCGGCCTGT ACAGCCTGAGCAGCGTCGTGACCGTGCCCA GCAGCAGCCTGGGCACCCAGACCTACATCT GCAACGTGAACCACAAGCCCAGCAACACAA AGGTGGACAAGCGGGTGGAACCCAAGAGCT GCGACAAGACCCACACCTGTCCCCCCTGCC CTGCCCCTGAAGCGGAGGGAGCCCCCTCC GTGTTCCTGTTCCCCCCAAAGCCTAAGGACA CCCTGATGATCAGCCGGACCCCCGAAGTGA CCTGCGTGGTGGTGGACGTGTCCCACGAGG ACCCTGAAGTGAAGTTTAATTGGTACGTGGA CGGCGTGGAAGTGCACAACGCCAAGACCAA GCCCAGAGAGGAACAGTACAACAGCACCTA CCGGGTGGTGTCCGTGCTGACCGTGCTGCA CCAGGACTGGCTGAACGGCAAAGAGTACAA GTGCAAGGTGTCCAACAAGGCCCTGCCTTC CTCCATCGAGAAAACCATCAGCAAGGCCAAA GGCCAGCCCCGCGAGCCCCAGGTGTACACA CTGCCCCCTAGCCGGGAAGAGATGACCAAG AACCAGGTGTCCCTGACCTGCCTCGTGAAG GGCTTCTACCCCAGCGACATTGCCGTGGAA TGGGAGAGCAACGGCCAGCCCGAGAACAAC TACAAGACCACCCCCCCTGTGCTGGACAGC GACGGCTCATTCTTCCTGTACAGCAAGCTGA CCGTGGACAAGAGCCGGTGGCAGCAGGGC AACGTGTTCAGCTGCTCCGTGATGCACGAG GCCCTGCACAACCACTACACCCAGAAGTCC CTGAGCCTGAGCCCCGGCAAG Light chain (DNA) SEQ ID NO: 57 CAGAGCGTGCTGACCCAGCCTCCTAGCGTG AGCGGTGCACCGGGCCAGCGCGTGACCATT AGCTGTAGCGGCAGCTCCTCCAATATTGGTA GCTATTACGTGAGCTGGTATCAGCAGCTGC CGGGCACGGCGCCGAAAGTTCTGATCTATC GTAATAATCAACGTCCTAGCGGCGTGCCGG ATCGCTTTAGCGGATCCAAAAGCGGCACCA GCGCCAGCCTGGCGATTACCGGCCTGCAAG CAGAAGATGAAGCGGATTATTACTGCGACAG CTGGGATCACAGCTCCATGAATGTTTTTGGC GGCGGTACCAAGCTGACCGTGCTGGGCCAG CCCAAAGCCGCCCCTAGCGTGACCCTGTTC CCCCCCTCGAGTGAGGAACTCCAGGCCAAC AAGGCCACCCTCGTGTGCCTGATCAGCGAC TTCTACCCTGGCGCCGTGACCGTGGCCTGG AAGGCCGATAGCAGCCCTGTGAAGGCCGGC GTGGAAACCACCACCCCCAGCAAGCAGAGC AACAACAAATACGCCGCCAGCAGCTACCTG AGCCTGACCCCCGAGCAGTGGAAGTCCCAC AGATCCTACAGCTGCCAGGTCACACACGAG GGCAGCACCGTGGAAAAGACCGTGGCCCCC ACCGAGTGCAGC MAB#1 DNA (optimized) HCDR1 (Kabat) SEQ ID NO: 70 AGCTACGCTATGCAC HCDR2 (Kabat) SEQ ID NO: 71 CGGATCAAGAGCAAGGCTCAAGGCGGCACC ACCGATTACGCCGCTCATGTGAAGGGC HCDR3 (Kabat) SEQ ID NO: 72 GTGTCCTTCTCCACCTTCGATGTG HCDR1 (Chothia) SEQ ID NO: 73 GGCTTCACCTTCTCCAGCTAC HCDR2 (Chothia) SEQ ID NO: 74 AAGAGCAAGGCTCAAGGCGGCACC HCDR3 (Chothia) SEQ ID NO: 72 GTGTCCTTCTCCACCTTCGATGTG LCDR1 (Kabat) SEQ ID NO: 75 TCCGGCTCCTCCTCCAACATCGGCTCCTACT ACGTGTCC LCDR2 (Kabat) SEQ ID NO: 76 CGGAACAACCAGCGGCCTTCT LCDR3 (Kabat) SEQ ID NO: 77 GACTCTTGGGACCACTCCTCCATGAACGTG LCDR1 (Chothia) SEQ ID NO: 75 TCCGGCTCCTCCTCCAACATCGGCTCCTACT ACGTGTCC LCDR2 (Chothia) SEQ ID NO: 76 CGGAACAACCAGCGGCCTTCT LCDR3 (Chothia) SEQ ID NO: 77 GACTCTTGGGACCACTCCTCCATGAACGTG VH SEQ ID NO: 78 GAAGTGCAGCTGGTGGAATCTGGCGGCGGA CTTGTGAAACCTGGCGGCTCTCTGAGACTGT CTTGTGCCGCTTCCGGCTTCACCTTCTCCAG CTACGCTATGCACTGGGTCCGACAGGCCCC TGGCAAAGGATTGGAGTGGGTCGGACGGAT CAAGAGCAAGGCTCAAGGCGGCACCACCGA TTACGCCGCTCATGTGAAGGGCAGATTCAC CATCTCTCGGGACGACTCCAAGAACACCCT GTACCTGCAGATGAACTCCCTGAAAACCGA GGACACCGCCGTGTACTACTGCGCCAGAGT GTCCTTCTCCACCTTCGATGTGTGGGGCCA GGGCACACTGGTTACAGTCTCGAGC VL SEQ ID NO: 79 CAGTCCGTGCTGACCCAGCCTCCTTCTGTTT CTGGTGCTCCTGGCCAGAGAGTGACCATCT CTTGCTCCGGCTCCTCCTCCAACATCGGCTC CTACTACGTGTCCTGGTATCAGCAGCTGCCT GGCACCGCTCCTAAGGTGCTGATCTACCGG AACAACCAGCGGCCTTCTGGCGTGCCCGAT AGATTCTCCGGCTCTAAGTCTGGCACCTCTG CCAGCCTGGCTATCACTGGACTGCAGGCTG AGGACGAGGCCGACTACTACTGCGACTCTT GGGACCACTCCTCCATGAACGTGTTCGGCG GAGGTACCAAGCTGACCGTGCTGGGACAG Heavy chain (IgG1) SEQ ID NO: 80 GAAGTGCAGCTGGTGGAATCTGGCGGCGGA IgG1 AEASS CTTGTGAAACCTGGCGGCTCTCTGAGACTGT CTTGTGCCGCTTCCGGCTTCACCTTCTCCAG CTACGCTATGCACTGGGTCCGACAGGCCCC TGGCAAAGGATTGGAGTGGGTCGGACGGAT CAAGAGCAAGGCTCAAGGCGGCACCACCGA TTACGCCGCTCATGTGAAGGGCAGATTCAC CATCTCTCGGGACGACTCCAAGAACACCCT GTACCTGCAGATGAACTCCCTGAAAACCGA GGACACCGCCGTGTACTACTGCGCCAGAGT GTCCTTCTCCACCTTCGATGTGTGGGGCCA

GGGCACACTGGTTACAGTCTCGAGCGCCTC CACCAAAGGACCCTCTGTGTTTCCTCTGGCT CCCTCCAGCAAGTCTACCTCTGGTGGAACA GCTGCCCTGGGCTGCCTGGTCAAGGATTAC TTTCCTGAGCCTGTGACCGTGTCCTGGAACT CTGGCGCTCTGACATCTGGCGTGCACACCT TTCCAGCTGTGCTGCAGTCCTCTGGCCTGTA CAGCCTGTCCTCTGTCGTGACCGTGCCTTCT AGCTCTCTGGGCACCCAGACCTACATCTGC AATGTGAACCACAAGCCTTCCAACACCAAGG TGGACAAGAGAGTGGAACCCAAGTCCTGCG ACAAGACCCACACCTGTCCTCCATGTCCTGC TCCAGAAGCTGAGGGCGCTCCTTCCGTGTT CCTGTTTCCTCCAAAGCCTAAGGACACCCTG ATGATCTCTCGGACCCCTGAAGTGACCTGC GTGGTGGTGGATGTGTCTCACGAGGACCCA GAAGTGAAGTTCAATTGGTACGTGGACGGC GTGGAAGTGCACAACGCCAAGACCAAGCCT AGAGAGGAACAGTACAACTCCACCTACAGA GTGGTGTCCGTGCTGACCGTGCTGCACCAG GATTGGCTGAACGGCAAAGAGTACAAGTGC AAGGTGTCCAACAAGGCCCTGCCTTCCAGC ATCGAAAAGACCATCTCCAAGGCCAAGGGC CAGCCTNGGGAACCCCAGGITTACACCCTG CCTCCAAGCCGGGAAGAGATGACCAAGAAC CAGGTGTCCCTGACCTGCCTCGTGAAGGGC TTCTACCCTTCCGATATCGCCGTGGAATGGG AGAGCAATGGCCAGCCTGAGAACAACTACA AGACAACCCCTCCTGTGCTGGACTCCGACG GCTCATTCTTCCTGTACTCCAAGCTGACAGT GGACAAGTCCAGATGGCAGCAGGGCAACGT GTTCTCCTGCTCCGTGATGCACGAGGCCCT GCACAATCACTACACACAGAAGTCCCTGTCT CTGTCCCCTGGCAAG Light chain (DNA) SEQ ID NO: 81 CAGTCCGTGCTGACCCAGCCTCCTTCTGTTT CTGGTGCTCCTGGCCAGAGAGTGACCATCT CTTGCTCCGGCTCCTCCTCCAACATCGGCTC CTACTACGTGTCCTGGTATCAGCAGCTGCCT GGCACCGCTCCTAAGGTGCTGATCTACCGG AACAACCAGCGGCCTTCTGGCGTGCCCGAT AGATTCTCCGGCTCTAAGTCTGGCACCTCTG CCAGCCTGGCTATCACTGGACTGCAGGCTG AGGACGAGGCCGACTACTACTGCGACTCTT GGGACCACTCCTCCATGAACGTGTTCGGCG GAGGTACCAAGCTGACCGTGCTGGGACAGC CTAAGGCTGCCCCTTCCGTGACACTGTTCCC TCCATCCTCTGAGGAACTGCAGGCCAACAA GGCTACCCTCGTGTGCCTGATCTCCGACTTT TACCCTGGCGCTGTGACCGTGGCCTGGAAG GCTGATAGTTCTCCTGTGAAGGCCGGCGTG GAAACCACCACACCTTCCAAGCAGTCCAACA ACAAATACGCCGCCTCCTCCTACCTGTCTCT GACCCCTGAACAGTGGAAGTCCCACCGGTC CTACAGCTGCCAAGTGACCCATGAGGGCTC CACCGTGGAAAAGACCGTGGCTCCTACCGA GTGCTCT

TABLE-US-00013 TABLE 2 Antibody sequences of MAB#2 MAB#2 SEQ ID NO: [aa] / DNA MAB#2 Protein HCDR1 (Kabat) SEQ ID NO: 27 SYAMH HCDR2 (Kabat) SEQ ID NO: 39 RIKSVAQGGTTDYAAHVKG HCDR3 (Kabat) SEQ ID NO: 40 VSHSTFDV HCDR1 (Chothia) SEQ ID NO: 30 GFTFSSY HCDR2 (Chothia) SEQ ID NO: 41 KSVAQGGT HCDR3 (Chothia) SEQ ID NO: 40 VSHSTFDV LCDR1 (Kabat) SEQ ID NO: 32 SGSSSNIGSYYVS LCDR2 (Kabat) SEQ ID NO: 33 RNNQRPS LCDR3 (Kabat) SEQ ID NO: 34 DSWDHSSMNV LCDR1 (Chothia) SEQ ID NO: 32 SGSSSNIGSYYVS LCDR2 (Chothia) SEQ ID NO: 33 RNNQRPS LCDR3 (Chothia) SEQ ID NO: 34 DSWDHSSMNV VH SEQ ID NO: 42 EVQLVESGGGLVKPGGSLRLSCAASGFTFSSY AMHWVRQAPGKGLEWVGRIKSVAQGGTTDY AAHVKGRFTISRDDSKNTLYLQMNSLKTEDTA VYYCARVSHSTFDVWGQGTLVTVSS VL SEQ ID NO: 43 QSVLTQPPSVSGAPGQRVTISCSGSSSNIGSY YVSWYQQLPGTAPKVLIYRNNQRPSGVPDRF SGSKSGTSASLAITGLQAEDEADYYCDSWDH SSMNVFGGGTKLTVLGQ Heavy chain SEQ ID NO: 44 EVQLVESGGGLVKPGGSLRLSCAASGFTFSSY IgG1_AEASS AMHWVRQAPGKGLEWVGRIKSVAQGGTTDY AAHVKGRFTISRDDSKNTLYLQMNSLKTEDTA VYYCARVSHSTFDVWGQGTLVTVSSASTKGP SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVT VPSSSLGTQTYICNVNHKPSNTKVDKRVEPKS CDKTHTCPPCPAPEAEGAPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDW LNGKEYKCKVSNKALPSSIEKTISKAKGQPREP QVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSK LTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK Light chain SEQ ID NO: 45 QSVLTQPPSVSGAPGQRVTISCSGSSSNIGSY YVSWYQQLPGTAPKVLIYRNNQRPSGVPDRF SGSKSGTSASLAITGLQAEDEADYYCDSWDH SSMNVFGGGTKLTVLGQPKAAPSVTLFPPSSE ELQANKATLVCLISDFYPGAVTVAWKADSSPV KAGVETTTPSKQSNNKYAASSYLSLTPEQWKS HRSYSCQVTHEGSTVEKTVAPTECS MAB#2 DNA HCDR1 (Kabat) SEQ ID NO: 58 AGCTATGCGATGCAC HCDR2 (Kabat) SEQ ID NO: 59 CGTATCAAATCCGTGGCCCAGGGCGGTACG ACCGACTACGCGGCGCACGTGAAAGGC HCDR3 (Kabat) SEQ ID NO: 60 GTTTCTCATTCCACTTTCGATGTT HCDR1 (Chothia) SEQ ID NO: 61 GGATTTACCTTCAGCAGCTAT HCDR2 (Chothia) SEQ ID NO: 62 AAATCCGTGGCCCAGGGCGGTACG HCDR3 (Chothia) SEQ ID NO: 60 GTTTCTCATTCCACTTTCGATGTT LCDR1 (Kabat) SEQ ID NO: 63 AGCGGCAGCTCCTCCAATATTGGTAGCTATT ACGTGAGC LCDR2 (Kabat) SEQ ID NO: 64 CGTAATAATCAACGTCCTAGC LCDR3 (Kabat) SEQ ID NO: 65 GACAGCTGGGATCACAGCTCCATGAATGTT LCDR1 (Chothia) SEQ ID NO: 63 AGCGGCAGCTCCTCCAATATTGGTAGCTATT ACGTGAGC LCDR2 (Chothia) SEQ ID NO: 64 CGTAATAATCAACGTCCTAGC LCDR3 (Chothia) SEQ ID NO: 65 GACAGCTGGGATCACAGCTCCATGAATGTT VH SEQ ID NO: 66 GAGGTGCAATTGGTGGAAAGCGGCGGTGGC CTGGTGAAACCAGGCGGCAGCCTGCGCCTG AGCTGCGCCGCCTCCGGATTTACCTTCAGC AGCTATGCGATGCACTGGGTGCGCCAGGCC CCGGGCAAAGGTCTCGAATGGGTGGGTCGT ATCAAATCCGTGGCCCAGGGCGGTACGACC GACTACGCGGCGCACGTGAAAGGCCGCTTT ACCATTAGCCGCGATGATTCGAAAAACACCC TGTATCTGCAAATGAACAGCCTGAAAACCGA AGATACGGCCGTGTATTATTGCGCGCGTGTT TCTCATTCCACTTTCGATGTTTGGGGCCAAG GCACCCTGGTGACTGTCTCGAGC VL SEQ ID NO: 67 CAGAGCGTGCTGACCCAGCCTCCTAGCGTG AGCGGTGCACCGGGCCAGCGCGTGACCATT AGCTGTAGCGGCAGCTCCTCCAATATTGGTA GCTATTACGTGAGCTGGTATCAGCAGCTGC CGGGCACGGCGCCGAAAGTTCTGATCTATC GTAATAATCAACGTCCTAGCGGCGTGCCGG ATCGCTTTAGCGGATCCAAAAGCGGCACCA GCGCCAGCCTGGCGATTACCGGCCTGCAAG CAGAAGATGAAGCGGATTATTACTGCGACAG CTGGGATCACAGCTCCATGAATGTTTTTGGC GGCGGTACCAAGCTGACCGTGCTGGGCCAG Heavy chain (IgG1) SEQ ID NO: 68 GAGGTGCAATTGGTGGAAAGCGGCGGTGGC IgG1 AEASS CTGGTGAAACCAGGCGGCAGCCTGCGCCTG AGCTGCGCCGCCTCCGGATTTACCTTCAGC AGCTATGCGATGCACTGGGTGCGCCAGGCC CCGGGCAAAGGTCTCGAATGGGTGGGTCGT ATCAAATCCGTGGCCCAGGGCGGTACGACC GACTACGCGGCGCACGTGAAAGGCCGCTTT ACCATTAGCCGCGATGATTCGAAAAACACCC TGTATCTGCAAATGAACAGCCTGAAAACCGA AGATACGGCCGTGTATTATTGCGCGCGTGTT TCTCATTCCACTTTCGATGTTTGGGGCCAAG GCACCCTGGTGACTGTCTCGAGCGCGTCGA CCAAAGGCCCCAGCGTGTTCCCTCTGGCCC CCAGCAGCAAGAGCACCTCTGGCGGAACAG CCGCCCTGGGCTGCCTGGTCAAGGACTACT TCCCCGAGCCCGTGACCGTGTCCTGGAACT CTGGCGCCCTGACCAGCGGCGTGCACACCT TTCCAGCCGTGCTCCAGAGCAGCGGCCTGT ACAGCCTGAGCAGCGTCGTGACCGTGCCCA GCAGCAGCCTGGGCACCCAGACCTACATCT GCAACGTGAACCACAAGCCCAGCAACACAA AGGTGGACAAGCGGGTGGAACCCAAGAGCT GCGACAAGACCCACACCTGTCCCCCCTGCC CTGCCCCTGAAGCGGAGGGAGCCCCCTCC GTGTTCCTGTTCCCCCCAAAGCCTAAGGACA CCCTGATGATCAGCCGGACCCCCGAAGTGA CCTGCGTGGTGGTGGACGTGTCCCACGAGG ACCCTGAAGTGAAGTTTAATTGGTACGTGGA CGGCGTGGAAGTGCACAACGCCAAGACCAA GCCCAGAGAGGAACAGTACAACAGCACCTA CCGGGTGGTGTCCGTGCTGACCGTGCTGCA CCAGGACTGGCTGAACGGCAAAGAGTACAA GTGCAAGGTGTCCAACAAGGCCCTGCCTTC CTCCATCGAGAAAACCATCAGCAAGGCCAAA GGCCAGCCCCGCGAGCCCCAGGTGTACACA CTGCCCCCTAGCCGGGAAGAGATGACCAAG AACCAGGTGTCCCTGACCTGCCTCGTGAAG GGCTTCTACCCCAGCGACATTGCCGTGGAA TGGGAGAGCAACGGCCAGCCCGAGAACAAC TACAAGACCACCCCCCCTGTGCTGGACAGC GACGGCTCATTCTTCCTGTACAGCAAGCTGA CCGTGGACAAGAGCCGGTGGCAGCAGGGC AACGTGTTCAGCTGCTCCGTGATGCACGAG GCCCTGCACAACCACTACACCCAGAAGTCC CTGAGCCTGAGCCCCGGCAAG Light chain (DNA) SEQ ID NO: 69 CAGAGCGTGCTGACCCAGCCTCCTAGCGTG AGCGGTGCACCGGGCCAGCGCGTGACCATT AGCTGTAGCGGCAGCTCCTCCAATATTGGTA GCTATTACGTGAGCTGGTATCAGCAGCTGC CGGGCACGGCGCCGAAAGTTCTGATCTATC GTAATAATCAACGTCCTAGCGGCGTGCCGG ATCGCTTTAGCGGATCCAAAAGCGGCACCA GCGCCAGCCTGGCGATTACCGGCCTGCAAG CAGAAGATGAAGCGGATTATTACTGCGACAG CTGGGATCACAGCTCCATGAATGTTTTTGGC GGCGGTACCAAGCTGACCGTGCTGGGCCAG CCCAAAGCCGCCCCTAGCGTGACCCTGTTC CCCCCCTCGAGTGAGGAACTCCAGGCCAAC AAGGCCACCCTCGTGTGCCTGATCAGCGAC TTCTACCCTGGCGCCGTGACCGTGGCCTGG AAGGCCGATAGCAGCCCTGTGAAGGCCGGC GTGGAAACCACCACCCCCAGCAAGCAGAGC AACAACAAATACGCCGCCAGCAGCTACCTG AGCCTGACCCCCGAGCAGTGGAAGTCCCAC AGATCCTACAGCTGCCAGGTCACACACGAG GGCAGCACCGTGGAAAAGACCGTGGCCCCC ACCGAGTGCAGC MAB#2 DNA OPTIMIZED HCDR1 (Kabat) SEQ ID NO: 82 AGCTACGCTATGCAC HCDR2 (Kabat) SEQ ID NO: 83 CGGATCAAGAGCGTTGCCCAAGGCGGCACC ACCGATTACGCTGCTCATGTGAAGGGC HCDR3 (Kabat) SEQ ID NO: 84 GTGTCCCACTCTACCTTCGATGTG HCDR1 (Chothia) SEQ ID NO: 85 GGCTTCACCTTCTCCAGCTAC HCDR2 (Chothia) SEQ ID NO: 86 AAGAGCGTTGCCCAAGGCGGCACC HCDR3 (Chothia) SEQ ID NO: 84 GTGTCCCACTCTACCTTCGATGTG LCDR1 (Kabat) SEQ ID NO: 87 TCCGGCTCCTCCTCCAACATCGGCTCCTACT ACGTGTCC LCDR2 (Kabat) SEQ ID NO: 88 CGGAACAACCAGCGGCCTTCT LCDR3 (Kabat) SEQ ID NO: 89 GACTCTTGGGACCACTCCTCCATGAACGTG LCDR1 (Chothia) SEQ ID NO: 87 TCCGGCTCCTCCTCCAACATCGGCTCCTACT ACGTGTCC LCDR2 (Chothia) SEQ ID NO: 88 CGGAACAACCAGCGGCCTTCT LCDR3 (Chothia) SEQ ID NO: 89 GACTCTTGGGACCACTCCTCCATGAACGTG VH SEQ ID NO: 90 GAAGTGCAGCTGGTGGAATCTGGCGGCGGA CTTGTGAAACCTGGCGGCTCTCTGAGACTGT CTTGTGCCGCTTCCGGCTTCACCTTCTCCAG CTACGCTATGCACTGGGTCCGACAGGCCCC TGGCAAAGGATTGGAGTGGGTCGGACGGAT CAAGAGCGTTGCCCAAGGCGGCACCACCGA TTACGCTGCTCATGTGAAGGGCAGATTCACC ATCAGCCGGGACGACTCCAAGAACACCCTG TACCTGCAGATGAACTCCCTGAAAACCGAG GACACCGCCGTGTACTACTGCGCCAGAGTG TCCCACTCTACCTTCGATGTGTGGGGCCAG GGCACACTGGTTACAGTCTCGAGC VL SEQ ID NO: 91 CAGTCCGTGCTGACCCAGCCTCCTTCTGTTT CTGGTGCTCCTGGCCAGAGAGTGACCATCT CTTGCTCCGGCTCCTCCTCCAACATCGGCTC CTACTACGTGTCCTGGTATCAGCAGCTGCCT GGCACCGCTCCTAAGGTGCTGATCTACCGG AACAACCAGCGGCCTTCTGGCGTGCCCGAT AGATTCTCCGGCTCTAAGTCTGGCACCTCTG CCAGCCTGGCTATCACTGGACTGCAGGCTG AGGACGAGGCCGACTACTACTGCGACTCTT GGGACCACTCCTCCATGAACGTGTTCGGCG GAGGTACCAAGCTGACCGTGCTGGGACAG Heavy chain (IgG1) SEQ ID NO: 92 GAAGTGCAGCTGGTGGAATCTGGCGGCGGA IgG1 AEASS CTTGTGAAACCTGGCGGCTCTCTGAGACTGT CTTGTGCCGCTTCCGGCTTCACCTTCTCCAG CTACGCTATGCACTGGGTCCGACAGGCCCC TGGCAAAGGATTGGAGTGGGTCGGACGGAT CAAGAGCGTTGCCCAAGGCGGCACCACCGA TTACGCTGCTCATGTGAAGGGCAGATTCACC ATCAGCCGGGACGACTCCAAGAACACCCTG TACCTGCAGATGAACTCCCTGAAAACCGAG GACACCGCCGTGTACTACTGCGCCAGAGTG TCCCACTCTACCTTCGATGTGTGGGGCCAG

GGCACACTGGTTACAGTCTCGAGCGCCTCC ACCAAAGGACCCTCTGTGTTTCCTCTGGCTC CCTCCAGCAAGTCTACCTCTGGTGGAACAG CTGCCCTGGGCTGCCTGGTCAAGGATTACT TTCCTGAGCCTGTGACCGTGTCCTGGAACTC TGGCGCTCTGACATCTGGCGTGCACACCTTT CCAGCTGTGCTGCAGTCCTCTGGCCTGTAC AGCCTGTCCTCTGTCGTGACCGTGCCTTCTA GCTCTCTGGGCACCCAGACCTACATCTGCA ATGTGAACCACAAGCCTTCCAACACCAAGGT GGACAAGAGAGTGGAACCCAAGTCCTGCGA CAAGACCCACACCTGTCCTCCATGTCCTGCT CCAGAAGCTGAGGGCGCTCCTTCCGTGTTC CTGTTTCCTCCAAAGCCTAAGGACACCCTGA TGATCTCTCGGACCCCTGAAGTGACCTGCG TGGTGGTGGATGTGTCTCACGAGGACCCAG AAGTGAAGTTCAATTGGTACGTGGACGGCG TGGAAGTGCACAACGCCAAGACCAAGCCTA GAGAGGAACAGTACAACTCCACCTACAGAG TGGTGTCCGTGCTGACCGTGCTGCACCAGG ATTGGCTGAACGGCAAAGAGTACAAGTGCA AGGTGTCCAACAAGGCCCTGCCTTCCAGCA TCGAAAAGACCATCTCCAAGGCCAAGGGCC AGCCTAGGGAACCCCAGGTTTACACCCTGC CTCCAAGCCGGGAAGAGATGACCAAGAACC AGGTGTCCCTGACCTGCCTCGTGAAGGGCT TCTACCCTTCCGATATCGCCGTGGAATGGGA GAGCAATGGCCAGCCTGAGAACAACTACAA GACAACCCCTCCTGTGCTGGACTCCGACGG CTCATTCTTCCTGTACTCCAAGCTGACAGTG GACAAGTCCAGATGGCAGCAGGGCAACGTG TTCTCCTGCTCCGTGATGCACGAGGCCCTG CACAATCACTACACACAGAAGTCCCTGTCTC TGTCCCCTGGCAAG Light chain (DNA) SEQ ID NO: 93 CAGTCCGTGCTGACCCAGCCTCCTTCTGTTT CTGGTGCTCCTGGCCAGAGAGTGACCATCT CTTGCTCCGGCTCCTCCTCCAACATCGGCTC CTACTACGTGTCCTGGTATCAGCAGCTGCCT GGCACCGCTCCTAAGGTGCTGATCTACCGG AACAACCAGCGGCCTTCTGGCGTGCCCGAT AGATTCTCCGGCTCTAAGTCTGGCACCTCTG CCAGCCTGGCTATCACTGGACTGCAGGCTG AGGACGAGGCCGACTACTACTGCGACTCTT GGGACCACTCCTCCATGAACGTGTTCGGCG GAGGTACCAAGCTGACCGTGCTGGGACAGC CTAAGGCTGCCCCTTCCGTGACACTGTTCCC TCCATCCTCTGAGGAACTGCAGGCCAACAA GGCTACCCTCGTGTGCCTGATCTCCGACTTT TACCCTGGCGCTGTGACCGTGGCCTGGAAG GCTGATAGTTCTCCTGTGAAGGCCGGCGTG GAAACCACCACACCTTCCAAGCAGTCCAACA ACAAATACGCCGCCTCCTCCTACCTGTCTCT GACCCCTGAACAGTGGAAGTCCCACCGGTC CTACAGCTGCCAAGTGACCCATGAGGGCTC CACCGTGGAAAAGACCGTGGCTCCTACCGA GTGCTCT

TABLE-US-00014 TABLE 3 Antibody sequences of RefMAB#1 SEQ ID RefMAB#1 NO: [aa] RefMAB#1 Protein Heavy SEQ ID EVQLVESGGGLVQPGGSLRLSCAASGFTFSS chain NO: 94 YVMHVVVRQATGKGLEVVVSAIDTGGGTYYA IgG1_AEASS DSVKGRFTISRENAKNSLYLQMNSLRAGDTA VYYCARDYYYYASGSYYKAFDIWGQGTMVT VSSASTKGPSVFPLAPSSKSTSGGTAALGCL VKDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNHKP SNTKVDKRVEPKSCDKTHTCPPCPAPEAEG APSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKEYKCKVSNKAL PSSIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENN YKTTPPVLDSDGSFFLYSKLTVDKSRWQQG NVFSCSVMHEALHNHYTQKSLSLSPGK Light SEQ ID EIVLTQSPGTLSLSPGERATLSCRASQSVSS chain NO: 95 RYLAVVYQQKPGQAPRLLIYGASSRATGIPD RFSGSGSGTDFTLTISRLEPEDFAVYYCQQY GSPLTFGQGTKLEIKRTVAAPSVFIFPPSDE QLKSGTASVVCLLNNFYPREAKVQWKVDNAL QSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC

WORKING EXAMPLES

Example 1: Antigen Generation and Quality Control

[0514] Amino acid sequences of C5aR and C5aR related GPCRs from various species were retrieved from publicly available sources (e.g. Uniprot), verified and produced in-house or by external service providers.

Synthetic Peptides

[0515] As antigens for the initial panning and screening, linear peptides covering the N-terminal extracellular region of human C5aR were used. The peptides were chemically synthesized with a biotin tag (JPT), RP-HPLC purified and delivered as lyophilized material. The lyophilized peptides were stored at -80.degree. C. Alternatively, the peptides were conjugated to Transferrin or bovine serum albumin (BSA).

TABLE-US-00015 TABLE 4 Amino acid sequence of N-terminal human C5aR peptide used for initial panning and screening. N-terminal C5aR peptides Human SEQ ID MDSFNYTTPDYGHYDDKDTLDLNTPVDKTSN NO: 12

[0516] As antigens for later binding studies, linear peptides comprising the N-terminal region of human and cynomolgus monkey C5aR were used. The peptides were chemically synthesized with a biotin tag (Genscript), RP-HPLC purified and delivered as lyophilized material. The lyophilized peptides were stored at -80.degree. C. For reconstitution, peptides were dissolved in the desired volume of PBS and stored at -80.degree. C.

TABLE-US-00016 TABLE 5 Amino acid sequences of N-terminal C5aR peptides used for binding studies. N-terminal C5aR peptides (C5aR_NT peptide) Human SEQ ID NO: 13 MDSFNYTTPDYGHYDDKDTLDLNTP VDKTSNTLRVPD Cynomolgus SEQ ID NO: 14 MDPFSSTTLDYEHYDGKNVLDSDTP VDKTSNTLRVPD

Recombinant Proteins

C1q Protein

[0517] C1q protein purified from pooled normal human plasma was purchased from Complement Technology, Inc. (Catalog #A099).

Human C5a Protein

[0518] Recombinant human C5a was either purchased from R&D Systems (CAT #: 2037-05) or was produced in house.

[0519] For in-house production, DNA encoding the amino acids of human C5a (Uniprot: P010311 Lys679-Arg751) was cloned into a pET21a expression vector (Novagen) in frame with an N-terminal ompA signal sequence followed by a sequence coding for maltose-binding protein (MBP), a FXa cleavage site and a GS linker.

[0520] Human C5a (hC5a) was expressed in E. coli BL21 (DE3) cells (Novagen) as a N-terminally tagged maltose-binding protein (MBP)-fusion protein. Protein expression was induced by the addition of IPTG and cultures were further cultivated for 20-23 h. Cells were harvested by centrifugation and the pellet was resuspended in lysis buffer (PBS buffer plus 2 mM MgCl.sub.2, 20 U/ml Benzonase (Roche) and 1 tablet/50 ml complete, EDTA-free protease inhibitor cocktail tablets (Roche)). Cells were disrupted either by chemical lysis or high-pressure homogenization. The resulting suspension was centrifuged and the supernatant was sterile filtered for further purification steps.

[0521] The hC5a-MBP-fusion protein was purified by Dextrin-Sepharose affinity chromatography using a MBP-Trap column (GE-Healthcare) and optionally polished by cation exchange chromatography using a Hi-Trap SP FF column (GE LifeSciences). The purified MBP-fusions were buffer-exchanged by PD10 columns (GE Healthcare) into FXa-digest buffer (20 mM Tris/HCl pH 8.0; 100 mM NaCl; 2 mM CaCl.sub.2)). The hC5a protein was released from the maltose-binding protein by addition of Factor Xa (1:100 (w/w)) and incubation 0/N in a rotary shaker at room temperature. The released hC5a was purified by cation exchange chromatography using a Hi-Trap SP FF column (GE LifeSciences). All affinity chromatography steps were performed using an AKTA Avant 25 preparative chromatography system.

[0522] Buffer exchange to PBS was performed using PD 10 columns (GE Healthcare). Samples were sterile filtered and hC5a concentration was determined by UV-spectrophotometry. The purity and integrity of the samples were analyzed in denaturing, reducing or non-reducing SDS-PAGE, SEC-HPLC and mass spectrometry.

Fc Gamma Receptors (Fc.gamma.R) and FcRn Receptors

[0523] DNA encoding the extracellular region of human Fc.gamma.RI, human Fc.gamma.RIIa (131H), human Fc.gamma.RIIa (131R), human Fc.gamma.RIIb, human Fc.gamma.RIIIa (158F) and human Fc.gamma.RIIIa (158V) were cloned in frame with an N-terminal V.sub.K leader sequence and a C-terminal 6.times.His-tag into a pMAX expression vector, which is a modified expression vector based on pcDNA3.1 (Thermo Fisher).

[0524] DNA encoding the extracellular region of human, cynomolgus, mouse or rat FcRn large subunit p51 was cloned in frame with an N-terminal V.sub.K leader sequence and a C-terminal AVI-6.times.His-tag into a pMAX expression vector, which is a modified expression vector based on pcDNA3.1 (Thermo Fisher). In addition, DNA encoding human, cynomolgus monkey, mouse or rat FcRn small subunit p14 (=identical with beta-2 microglobulin) protein was cloned into a second open reading frame in frame with an N-terminal Vk leader sequence. The amino acid sequences of the produced receptors are summarized in Table 6 and 7.

[0525] The HEK293-6E cell line was developed by the National Research Council of Canada (NRC). Cells were maintained in Freestyle F17 medium (Thermo Scientific) in a humidified CO2-incubator at 37.degree. C. and 6% CO.sub.2. HKB11 (Parental clone: U.S. Pat. No. 6,136,599. J. Biomed. Sci. 2002; 9:631-638) is a human hybrid cell line resulting from a fusion of HEK293 human embryonic kidney and 2B8 Burkitt lymphoma cells. HKB11 #52 cells were maintained in MAC1.0 medium containing 1% FCS in a humidified CO.sub.2 incubator at 37.degree. C. and 6% CO.sub.2.

[0526] HKB11#52 or HEK293-6E cells were transiently transfected one day post seeding with a commercially available transfection reagent according to the manufacturer's instructions. The cells were cultured for 3 days and the conditioned cell culture supernatant was harvested by centrifugation followed by sterile filtration (0.22 .mu.m). Stably transfected HKB11#52 pools were generated by transfection of cells followed by selection with 800 .mu.g/mL G418 (Thermo Scientific). Expression of antigens from stable pools was done for 4 days post seeding. The conditioned cell culture supernatants were harvested by centrifugation followed by sterile filtration (0.22 .mu.m).

[0527] The respective proteins were purified by IMAC using Protino Ni-NTA columns (Macherey-Nagel). All chromatography steps were performed using AKTA chromatography systems (GE Healthcare). The samples were buffer-exchanged to D-PBS using PD10 columns (GE Healthcare). In some cases, a polishing preparative SEC step was performed in D-PBS using a Superdex 200 column (GE Healthcare).

[0528] Biotinylation of FcRn heterodimers was performed by in vitro biotinylation using the BirA Kit (Avidity) followed by a preparative SEC using a Superdex 200 column (GE Healthcare).

[0529] The quality of the samples was analyzed by denaturing, reducing or non-reducing SDS-PAGE, Streptavidin-Shift Assay, HP-SEC and DLS.

TABLE-US-00017 TABLE 6 Amino acid sequences of produced FcRn proteins. SEQ ID FcRn Fusion Protein UniProt IDs: NO: Protein sequence Human FcRn p51 [1- P55899 (p51) SEQ ID AESHLSLLYHLTAVSSPAPGTPAFWVSGWLGPQ 297] / AviHis / p14 P61769 (p14) NO: 15 QYLSYNSLRGEAEPCGAWVWENQVSWYWEKE biotinylated TTDLRIKEKLFLEAFKALGGKGPYTLQGLLGCEL GPDNTSVPTAKFALNGEEFMNFDLKQGTWGGD WPEALAISQRWQQQDKAANKELTFLLFSCPHRL REHLERGRGNLEWKEPPSMRLKARPSSPGFSV LTCSAFSFYPPELQLRFLRNGLAAGTGQGDFGP NSDGSFHASSSLTVKSGDEHHYCCIVQHAGLAQ PLRVELESPAKSSVNSRGLNDIFEAQKIEWHEHH HHHHIQRTPKIQVYSRHPAENGKSNFLNCYVSG FHPSDIEVDLLKNGERIEKVEHSDLSFSKDWSFY LLYYTEFTPTEKDEYACRVNHVTLSQPKIVKWDR DM Cynomolgus FcRn p51 Q8SPV9 (p51) SEQ ID AESHLSLLYHLTAVSSPAPGTPAFVVVSGWLGPQ [1-297]_AviHis / p14 P61769 (p14) NO: 16 QYLSYDSLRGQAEPCGAWVWENQVSWYWEKE biotinylated TTDLRIKEKLFLEAFKALGGKGPYTLQGLLGCEL SPDNTSVPTAKFALNGEEFMNFDLKQGTWGGD WPEALAISQRWQQQDKAANKELTFLLFSCPHRL REHLERGRGNLEWKEPPSMRLKARPGNPGFSV LTCSAFSFYPPELQLRFLRNGMAAGTGQGDFGP NSDGSFHASSSLTVKSGDEHHYCCIVQHAGLAQ PLRVELETPAKSSVNSRGLNDIFEAQKIEWHEHH HHHHIQRTPKIQVYSRHPPENGKPNFLNCYVSG FHPSDIEVDLLKNGEKMGKVEHSDLSFSKDWSF YLLYYTEFTPNEKDEYACRVNHVTLSGPRTVKW DRDM Mouse FcRn p51 [1- Q61559 (p51) SEQ ID SETRPPLMYHLTAVSNPSTGLPSFWATGWLGP 297] / AviHis / p14 P61769 (p14) NO: 17 QQYLTYNSLRQEADPCGAWMWENQVSWYWEK biotinylated ETTDLKSKEQLFLEALKTLEKILNGTYTLQGLLGC ELASDNSSVPTAVFALNGEEFMKFNPRIGNWTG EWPETEIVANLWMKQPDAARKESEFLLNSCPER LLGHLERGRRNLEWKEPPSMRLKARPGNSGSS VLTCAAFSFYPPELKFRFLRNGLASGSGNCSTG PNGDGSFHAWSLLEVKRGDEHHYQCQVEHEGL AQPLTVDLDSSARSSVNSRGLNDIFEAQKIEWHE HHHHHHIQKTPQIQVYSRHPPENGKPNILNCYVT QFHPPHIEIQMLKNGKKIPKVEMSDMSFSKDWS FYILAHTEFTPTETDTYACRVKHDSMAEPKTVYW DRDM Rat FcRn p51 [1- P13599 (p51) SEQ ID AEPRLPLMYHLAAVSDLSTGLPSFWATGWLGAQ 298] / AviHis / p14 P61769 (p14) NO: 96 QYLTYNNLRQEADPCGAWIWENQVSWYWEKET biotinylated TDLKSKEQLFLEAIRTLENQINGTFTLQGLLGCEL APDNSSLPTAVFALNGEEFMRFNPRTGNWSGE WPETDIVGNLWMKQPEAARKESEFLLTSCPERL LGHLERGRQNLEWKEPPSMRLKARPGNSGSSV LTCAAFSFYPPELKFRFLRNGLASGSGNCSTGP NGDGSFHAWSLLEVKRGDEHHYQCQVEHEGLA QPLTVDLDSPARSSVNSRGLNDIFEAQKIEWHEH HHHHHIQKTPQIQVYSRHPPENGKPNFLNCYVS QFHPPQIEIELLKNGKKIPNIEMSDLSFSKDWSFY ILAHTEFTPTETDVYACRVKHVTLKEPKTVTWDR DM

TABLE-US-00018 TABLE 7 Amino acid sequences of produced human Fc.gamma.R proteins. UniProt SEQ ID Fc.gamma.R Fusion Protein IDs: NO: Protein sequence Human Fc.gamma.RI (1- P12314 SEQ ID QVDTTKAVITLQPPWVSVFQEETVTLHCEVLHLP 279)_HIS.sub.6 NO: 18 GSSSTQWFLNGTATQTSTPSYRITSASVNDSGE YRCQRGLSGRSDPIQLEIHRGWLLLQVSSRVFT EGEPLALRCHAWKDKLVYNVLYYRNGKAFKFFH WNSNLTILKTNISHNGTYHCSGMGKHRYTSAGIS VTVKELFPAPVLNASVTSPLLEGNLVTLSCETKLL LQRPGLQLYFSFYMGSKTLRGRNTSSEYQILTAR REDSGLYWCEAATEDGNVLKRSPELELQVNSR HHHHHH Human Fc.gamma.RIIa (1- P12318 SEQ ID QAAAPPKAVLKLEPPWINVLQEDSVTLTCQGAR 209)-(131H)- NO: 19 SPESDSIQWFHNGNLIPTHTQPSYRFKANNNDS HIS.sub.6(133-2) GEYTCQTGQTSLSDPVHLTVLSEWLVLQTPHLE FQEGETIMLRCHSWKDKPLVKVTFFQNGKSQKF SHLDPTFSIPQANHSHSGDYHCTGNIGYTLFSSK PVTITVQVPSVNSRHHHHHH Human Fc.gamma.RIIa (1- P12318 SEQ ID QAAAPPKAVLKLEPPWINVLQEDSVTLICQGAR 209)-(131R)-HIS.sub.6 NO: 20 SPESDSIQWFHNGNLIPTHTQPSYRFKANNNDS (137-3) GEYTCQTGQTSLSDPVHLTVLSEWLVLQTPHLE FQEGETIMLRCHSWKDKPLVKVTFFQNGKSQKF SRLDPTFSIPQANHSHSGDYHCTGNIGYTLFSSK PVTITVQVPSVNSRHHHHHH Human Fc.gamma.RIIIa (1- P08637 SEQ ID GMRTEDLPKAVVFLEPQWYRVLEKDSVTLKCQG 193)-(158F)-HIS.sub.6 NO: 21 AYSPEDNSTQWFHNESLISSQASSYFIDAATVDD (141-2) SGEYRCQTNLSTLSDPVQLEVHIGWLLLQAPRW VFKEEDPIHLRCHSWKNTALHKVTYLQNGKGRK YFHHNSDFYIPKATLKDSGSYFCRGLFGSKNVSS ETVNITITQGVNSRHHHHHH Human Fc.gamma.RIIIa (1- P08637 SEQ ID GMRTEDLPKAVVFLEPQWYRVLEKDSVTLKCQG 193)-(158V)-HIS.sub.6 NO: 22 AYSPEDNSTQWFHNESLISSQASSYFIDAATVDD SGEYRCQTNLSTLSDPVQLEVHIGWLLLQAPRW VFKEEDPIHLRCHSWKNTALHKVTYLQNGKGRK YFHHNSDFYIPKATLKDSGSYFCRGLVGSKNVS SETVNITITQGVNSRHHHHHH Human Fc.gamma.RIIB (1- P31994 SEQ ID TPAAPPKAVLKLEPQWINVLQEDSVTLTCRGTHS 202)-HIS.sub.6 NO: 23 PESDSIQWFHNGNLIPTHTQPSYRFKANNNDSG EYTCQTGQTSLSDPVHLTVLSEWLVLQTPHLEF QEGETIVLRCHSWKDKPLVKVTFFQNGKSKKFS RSDPNFSIPQANHSHSGDYHCTGNIGYTLYSSK PVTITVQAPSDNSRHHHHHH

Virus-Like-Particles (VLPs)

[0530] VLPs stably expressing either one of the two natural variants of human C5aR (D/K variant or N/N variant) as well as mouse C5aR were generated in house as described in WO 2015/193143. All cloning experiments were performed using standard technologies. Antigens of interest were cloned in a suited two vector system for the expression in mammalian cells. In this system, one vector expresses GAG and the other vector expresses the GPCR-GAG fusion protein. Expression in these vectors is under the control of the CMV promoter. Subsequently, host cells were transfected with the two vectors. The generated constructs were produced as fusion proteins, in which the antigen of interest is fused N-terminal to the GAG-protein (HV1B1 (Uni-Prot ID: P03347)). Expression of the proteins and production of the VLPs was done under standard conditions in suspension cultures. Host cells used in the present experiments were HKB11 cells (ATCC; CRL-12568) and HEK cells (Life Technologies). Three days post transfection the supernatants containing the VLPs were harvested and purified using standard procedures (including precipitation and ion exchange chromatography). The proteins isolated were subjected to SDS-PAGE chromatography. The supernatants were probed with a commercial anti-GAG antibody or a commercial anti-C5aR antibody and revealed that co-expression of GAG and a GPCR-GAG fusion protein resulted in a high expression level of GAG and GPCR-GAG in the VLPs. This confirmed that the C5aR antigens were efficiently integrated into the VLPs, and that the C5aR antigens were detectable with antibodies. The amino acid sequences of the produced fusion proteins are summarized in Table 8.

TABLE-US-00019 TABLE 8 Protein sequences of C5aR-GAG fusion protein expressed on virus-like-particles. C5aR-HV1B1 fusion SEQ ID protein NO: Protein sequence HIS.sub.6-human C5aR SEQ ID DGSHHHHHHGTMDSFNYTTPDYGHYDDKDTLDLNTPVDKTSNT (D/K variant)-HV1B1- NO: 24 LRVPDILALVIFAVVFLVGVLGNALVVWVTAFEAKRTINAIWFLNLA GAG VADFLSCLALPILFTSIVQHHHWPFGGAACSILPSLILLNMYASILL LATISADRFLLVFKPIWCQNFRGAGLAWIACAVAWGLALLLTIPSF LYRVVREEYFPPKVLCGVDYSHDKRRERAVAIVRLVLGFLWPLL TLTICYTFILLRTWSRRATRSTKTLKVVVAVVASFFIFWLPYQVTGI MMSFLEPSSPTFLLLKKLDSLCVSFAYINCCINPIIYVVAGQGFQG RLRKSLPSLLRNVLTEESVVRESKSFTRSTVDTMAQKTQAVDIDY KDDDDKIEGRMDGARASVLSGGELDRWEKIRLRPGGKKKYKLK HIVWASRELERFAVNPGLLETSEGCRQILGQLQPSLQTGSEELR SLYNTVATLYCVHQRIEIKDTKEALDKIEEEQNKSKKKAQQAAAD TGHSSQVSQNYPIVQNIQGQMVHQAISPRTLNAWVKVVEEKAFS PEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEA AEWDRVHPVHAGPIAPGQMREPRGSDIAGTTSTLQEQIGWMTN NPPIPVGEIYKRWIILGLNKIVRMYSPTSILDIRQGPKEPFRDYVDR FYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPAATL EEMMTACQGVGGPGHKARVLAEAMSQVTNTATIMMQRGNFRN QRKMVKCFNCGKEGHTARNCRAPRKKGCWKCGKEGHQMKDC TERQANFLGKIWPSYKGRPGNFLQSRPEPTAPPFLQSRPEPTAP PEESFRSGVETTTPPQKQEPIDKELYPLTSLRSLFGNDPSSQVN SRGLNDIFEAQKIEWHE HIS.sub.6--human C5aR SEQ ID DGSHHHHHHGTMNSFNYTTPDYGHYDDKDTLDLNTPVDKTSNT (N/N variant)-HV1B1- NO: 25 LRVPDILALVIFAVVFLVGVLGNALVVWVTAFEAKRTINAIWFLNLA GAG VADFLSCLALPILFTSIVQHHHWPFGGAACSILPSLILLNMYASILL LATISADRFLLVFKPIWCQNFRGAGLAWIACAVAWGLALLLTIPSF LYRVVREEYFPPKVLCGVDYSHDKRRERAVAIVRLVLGFLWPLL TLTICYTFILLRTWSRRATRSTKTLKVVVAVVASFFIFWLPYQVTGI MMSFLEPSSPTFLLLNKLDSLCVSFAYINCCINPIIYVVAGQGFQG RLRKSLPSLLRNVLTEESVVRESKSFTRSTVDTMAQKTQAVDIGA RASVLSGGELDRWEKIRLRPGGKKKYKLKHIVWASRELERFAVN PGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTVATLYCVHQRI EIKDTKEALDKIEEEQNKSKKKAQQAAADTGHSSQVSQNYPIVQ NIQGQMVHQAISPRTLNAWVKVVEEKAFSPEVIPMFSALSEGAT PQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRVHPVHAGPI APGQMREPRGSDIAGTTSTLQEQIGWMTNNPPIPVGEIYKRWIIL GLNKIVRMYSPTSILDIRQGPKEPFRDYVDRFYKTLRAEQASQEV KNWMTETLLVQNANPDCKTILKALGPAATLEEMMTACQGVGGP GHKARVLAEAMSQVTNTATIMMQRGNFRNQRKMVKCFNCGKE GHTARNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPS YKGRPGNFLQSRPEPTAPPFLQSRPEPTAPPEESFRSGVETTTP PQKQEPIDKELYPLTSLRSLFGNDPSSQ HIS.sub.6--mouse C5aR- SEQ ID DGSHHHHHHGTMDPIDNSSFEINYDHYGTMAPNIPADGIHLPKR HV1B1-GAG NO: 26 QPGDVAALIIYSVVFLVGVPGNALWWVTAFEARRAVNAIWFLNL AVADLLSCLALPVLFTTVLNHNYWYFDATACIVLPSLILLNMYASIL LLATISADRFLLVFKPIWCQKVRGTGLAWMACGVAVVVLALLLTIP SFVYREAYKDFYSEHTVCGINYGGGSFPKEKAVAILRLMVGFVLP LLTLNICYTFLLLRTWSRKATRSTKTLKVVMAVVICFFIFWLPYQV TGVMIAWLPPSSPTLKRVEKLNSLCVSLAYINCCVNPIIYVMAGQ GFHGRLLRSLPSIIRNALSEDSVGRDSKTFTPSTTDTSTRKSQAV DIDYKDDDDKIEGRMDGARASVLSGGELDRWEKIRLRPGGKKKY KLKHIVWASRELERFAVNPGLLETSEGCRQILGQLQPSLQTGSE ELRSLYNTVATLYCVHQRIEIKDTKEALDKIEEEQNKSKKKAQQA AADTGHSSQVSQNYPIVQNIQGQMVHQAISPRTLNAWVKVVEEK AFSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETIN EEAAEWDRVHPVHAGPIAPGQMREPRGSDIAGTTSTLQEQIGW MTNNPPIPVGEIYKRWIILGLNKIVRMYSPTSILDIRQGPKEPFRDY VDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPA ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNTATIMMQRGN FRNQRKMVKCFNCGKEGHTARNCRAPRKKGCWKCGKEGHQM KDCTERQANFLGKIWPSYKGRPGNFLQSRPEPTAPPFLQSRPE PTAPPEESFRSGVETTTPPQKQEPIDKELYPLTSLRSLFGNDPSS QVNSRGLNDIFEAQKIEWHE

Cell Lines

[0531] CHO Flp-In cells stably expressing full length human C5aR, cynomolgus C5aR, mouse C5aR, rat C5aR and the C5aR related GPCRs human C5L2, human C3aR, human FPR1 and human ChemR23 were generated. For the generation of Flp-In CHO cells, various vector constructs were gene synthesized in-house and transfection of cells was performed according to the instructor's manual (Thermofischer/Invitrogen). All constructs contained an N-terminal V5/His tag. Commercially available anti-His (e.g. Dianova CAT #DIA-910) or anti-V5 (e.g. AbD Serotec CAT #MCA2285GA) detection antibodies were used to confirm the expression of the respective GPCR on the surface of the cell line, even in the absence of a commercially available specific anti-GPCR tool antibody.

Example 2: Generation of Human C5aR Specific Antibodies from the HuCAL PLATINUM.RTM. Library

[0532] For antibody generation, the HuCAL Platinum.RTM. library was used to select for antibodies with specificity for human C5aR. The HuCAL PLATINUM.RTM. library is a phagemid library based on the HuCAL concept (Knappik et al., (2000) J Mol Biol 296:57-86) and employs the CysDisplay.RTM. technology for displaying the Fab on the phage surface (Lohning et al., WO2001/05950).

[0533] To identify human C5aR specific antibodies, panning strategies were performed using human and cynomolgus monkey C5aR antigen material to select species cross-reactive antibodies. Each conducted panning comprised of at least 3 individual rounds of selection against various C5aR antigens (either as soluble recombinant antigens or overexpressed on cells).

[0534] Although the overall homology between cynomolgus and human C5aR with 90% is rather high, the extracellular domains share only 75% identity of the protein sequences. Indeed, the identification of cynomolgus C5aR cross-reactive antibodies turned out to be challenging, with the ancestor antibody of MAB#1 and MAB#2 being one of a few candidates revealing specific cell binding to human C5aR expressed on cells, cross-reactivity to cynomolgus monkey C5aR and no binding to other related GPCRs.

[0535] Bead based solution pannings against peptides representing the N-terminus (NT) of human C5aR were conducted which resulted in the identification of the ancestor antibody of MAB#1 and MAB#2 with specificity for human and cynomolgus monkey C5aR and being able to bind to full-length human C5aR expressed on cells.

[0536] This clone was subjected to two consecutively conducted affinity maturation panning using CHO Flp-In cells engineered to overexpress either human or cynomolgus C5aR in order to further increase affinity and specificity for human and cynomolgus C5aR. In addition, antibody engineering was conducted to further increase specificity, to remove potential posttranslational modification sites (PTM motifs) and for germlining purposes.

[0537] With this last step of engineering process, MAB#1 and MAB#2 were identified as potential therapeutic candidates. Since both candidates, MAB#1 and MAB#2 are derived from the same ancestors, they share similar amino acid sequences and in vitro characteristics.

These two antibodies are further described in the examples as outlined below.

Example 3: Production of Human C5aR Specific Antibodies

[0538] Both, MAB#1 and MAB#2 are of the human IgG1f isotype but are engineered in the Fc region to abolish the ability of the antibodies to mediate immune effector function. The Fc region comprises 5 amino acid substitutions compared to the wild-type human IgG1 Fc region, namely L234A, L235E, G237A, A330S and P331S (h_IgG1f_AEASS) with numbering according EU index.

[0539] The antibodies consist of the heavy chain framework VH3-15 and the antibody light chain framework lambda 1.

Transient Antibody Production--Advanced Micro Scale Production HKB11

[0540] Eukaryotic HKB11#52 cells were transiently transfected with mammalian expression vectors encoding heavy and light chains of MAB#1 or MAB#2 (human IgG1_AEASS), respectively. Cell culture supernatants were harvested 7 days post transfection and subjected to Protein A affinity chromatography (MabSelect SuRe|GE Healthcare) using a liquid handling station. The samples remained in neutralized elution buffer (NaPS: 137 mM NaPhosphate, 81 mM NaCl, pH 7). Samples were sterile filtered (0.2 .mu.m pore size). Protein concentrations were determined by UV-spectrophotometry at 280 .mu.m and purity of IgG was analysed under denaturing, reducing conditions using CE-SDS (LabChip GXII|Perkin Elmer). UHP-SEC was performed to analyse IgG preparations in native state.

Transient Antibody Production--Exploratory Scale Production in CHO

[0541] CHO3-E7 cells were transiently transfected with mammalian expression vector encoding heavy and light chains of MAB#1 or MAB#2 (human IgG1_AEASS), respectively. Cell culture supernatants were harvested on day 6 post transfection and subjected to standard Protein A affinity chromatography (MabSelect SuRe|GE Healthcare). If not stated otherwise, buffer exchange was performed to 1.times.Dulbecco's PBS (pH 7.2|Invitrogen) and samples were sterile filtered (0.2 .mu.m pore size). Protein concentrations were determined by UV-spectrophotometry at 280 nm and purity of IgG was analysed under denaturing, reducing and non-reducing conditions using CE-SDS (LabChip GXII|Perkin Elmer). UHP-SEC was performed to analyse IgG preparations in native state.

Results Transient Production

[0542] Data on product quality (SEC monomer content) and productivity of MAB#1 and MAB#2 are summarized in Table 9. Overall, acceptable monomer contents (>95%) and yields (>55 mg/L in HKB11 cells) were achieved. Volumetric yields derived from CHO3E-7 transient Exploratory Scale productions were also in the expected range.

TABLE-US-00020 TABLE 9 Production data of MAB#1 and MAB#2 in transient expression SEC Volumetric Antibody Production Platform Monomer % yield [mg/L] MAB#1 Advanced Micro HKB11 97.5 56.1 Exploratory CHO 96.4 3.0 MAB#2 Advanced Micro HKB11 97.4 58.9 Exploratory CHO 98.5 2.4

Generation of Stable HKB11 Pools

[0543] For the generation of HKB11#52 pools stably expressing MAB#1 or MAB#2, a two-vector system was used for co-transfection.

[0544] In order to facilitate pool selection these vectors contain Zeocin and Neomycin resistance cassettes. The two vectors were transiently co-transfected into actively dividing HKB11#52 cells in a 1:1 ratio. One day post transfection selection was started by the addition of 160 .mu.g/mL Zeocin and 800 .mu.g/mL Geneticin to the cell suspension. During the selection cell count and viability initially decreased. 20 days after transfection cells started to recover. Reaching a viability of .about.80%, stable pools were scaled up to the desired volume depending on the amount needed. Cell culture supernatants of the batch productions were harvested on day 6 post seeding.

Large Scale Purification of MAB#1 and MAB#2

[0545] Purification of MAB#1 and MAB#2 from cell culture supernatants of HKB11#52 stable pools via Protein A affinity chromatography (MabSelect SURE|GE Healthcare) was performed using 100 mM Citrate, 150 mM NaCl pH 3.5 as elution buffer. After incubation at pH 3.5 for 60 minutes, the samples were neutralized. Buffer exchange was performed into 150 mM Histidine, pH 6.0 and samples were sterile filtered (0.2 .mu.m pore size). Protein concentrations were determined by UV-spectrophotometry at 280 nm and purity of IgG was analysed under denaturing, reducing and non-reducing conditions using CE-SDS (LabChip GXII|Perkin Elmer). UHP-SEC was performed to analyse IgG preparations in native state.

Results Large Scale Production

[0546] As summarized in Table 10, production of MAB#1 and MAB#2 resulted in favorable yields, purity and integrity.

TABLE-US-00021 TABLE 10 Production data of MAB#1 and MAB#2 derived from stable cell pool expressions SEC Volumetric Production Monomer HMW LMW yield Antibody Platform [%] [%] [%] [mg/L] MAB#1 Large Scale HKB11 99.1 0.5 0.4 60.5 MAB#2 Large Scale HKB11 99.1 0.6 0.3 100.5

Control Antibodies

[0547] Various control antibodies were produced and included in experiments for comparative purposes:

RefMAB#1: Benchmark Antibody

[0548] Nucleotide sequences encoding the VH and VL region from the human C5aR specific antibody "IPH5401" were retrieved from US patent application US2013/0295116 (NOVO NORDISK-clone 32F3A6GL). Nucleotide sequences were gene synthesized as linear DNA fragments with appropriate flanking regions (e.g. suitable restriction enzyme recognition sites, linker sequences) either in-house or by an external provider. The DNA fragments were cloned into suited mammalian IgG expression vectors encoding heavy and light chains of human IgG1_AEASS as described above by using standard molecular biology methods. RefMAB#1 was transiently produced as described above. The heavy and light chain amino acid sequences of RefMAB#1 are depicted in Table 3.

Additional Control Antibodies:

[0549] An in-house negative isotype control antibody with specificity for hen-egg lysozyme (MOR03207) as well as an in-house positive control antibody (anti-C5aR antibody) with specificity for the N-terminus of human C5aR were transiently produced as described above either in human IgG1 or human IgG1_AEASS format.

[0550] Characterization of the Binding Properties of MAB#1 and MAB#2

Example 4: Monovalent Affinity Determination for MAB#1 and MAB#2 for C5aR N-Terminal Peptides Using SPR

Method

[0551] K.sub.D determination via IgG capture setup was performed at 25.degree. C. with a Biacore T200 instrument (Biacore, GE Healthcare). Approx. 500 RU of IgG diluted in HBS-EP+, pH 7.4, were captured on a CM5 chip (Biacore, GE Healthcare) immobilised with anti-human-Fc antibody (GE Healthcare) using standard EDC-NHS amine coupling chemistry. The reference flow cell 1 was only activated and deactivated. Kinetic measurements were done using 6 different human or cynomolgus C5aR_NT-bio peptide concentrations (SEQ ID NO: 13 or SEQ ID NO: 14, respectively) (2n serial dilution, 2000 to 62.5 nM) with HBS-EP+(GE Healthcare) as running buffer (injection time 300 s; dissociation time 600 s; flow rate 30 .mu.L/min). After each cycle the sensor chip was regenerated to remove bound peptide/antibody complex with 3.times.20 s injections of 3 mM MgCl.sub.2. A blank injection of running buffer was used for double referencing. All sensorgrams were fitted using Biacore T200 Evaluation Software 3.1, (Biacore, GE Healthcare) to determine k.sub.on and k.sub.off rate constants, which were used to calculate K.sub.D. The raw data was fitted with a 1:1 binding model, with parameters R.sub.max set to local and RI set to 0.

Results

[0552] Results are summarized in Table 11. Both antibodies revealed similar binding to the human C5aR peptide with K.sub.D values in the low double-digit nanomolar range and weaker binding to the cynomolgus C5aR peptide.

TABLE-US-00022 TABLE 11 Determination of association and dissociation rate constants for binding of MAB#1 and MAB#2 to human and cynomolgus C5aR N-terminal peptides. Antibody Antigen k.sub.on [1/Ms] k.sub.off [1/s] K.sub.D [nM] Comment MAB#2 Human 4.33E+04 1.27E-03 29 MAB#1 C5aR_NT 5.01E+04 1.79E-03 36 MAB#2 Cynomolgus 4.29E+03* 1 45E-01* 34000* fast k.sub.on, C5aR_NT fast k.sub.off MAB#1 1.36E+04* 1.11E-01 8200* fast k.sub.on, fast k.sub.off *Values formatted in grey italics are intended for ranking purposes.

Example 5: Apparent Affinity (Bivalent) Determination of MAB#1 and MAB#2 for C5aR N-Terminal Peptides Using Octet

Method

[0553] Apparent K.sub.D determination via C5aR_NT-bio peptide capture setup was performed at 27.degree. C. with the Octet HTX instrument (ForteBIO, Pall Life Sciences). Approx. 0.03 nm of peptide diluted in PBS, pH 7.4, were loaded onto streptavidin (SA) sensors (ForteBIO, Pall Life Sciences). Kinetic measurements were done using 7 different IgG concentrations (3-fold serial dilution, 200 to 0.27 nM for human C5aR_NT-bio peptide (SEQ ID NO: 13) and 1000 to 1.4 nM for cynomolgus C5aR_NT-bio peptide (SEQ ID NO: 14) in Octet buffer (PBS, 0.05% (v/v) Tween-20, 0.1% (w/v) BSA) with 480 s association time and 900 s dissociation time. After each dissociation step the sensors were regenerated to remove bound antibody (3.times.30 s Gly/HCl, pH 1.5). All sensorgrams were fitted using Octet Data Analysis Software 10.0 (ForteBio) to determine apparent k.sub.om and app. k.sub.off rate constants, which were used to calculate apparent K.sub.D (using a 1:1 binding model).

Results

[0554] Results are summarized in Table 12. Both antibodies revealed strong binding to the human C5aR peptide in bivalent format with apparent K.sub.D values in the double to triple digit picomolar range. Binding to the cynomolgus C5aR peptide was approx. 1000-fold weaker. The observed weaker binding to cynomolgus monkey C5aR peptide appeared mainly due to fast Ice rates. In terms of apparent affinities to human C5aR_NT, MAB#2 exhibited an about 10-fold weaker binding affinity compared to MAB#1 (K.sub.D: 290 pM vs. 20 pM).

TABLE-US-00023 TABLE 12 Determination of apparent association and dissociation rate constants for binding of MAB#1 and MAB#2 to human and cynomolgus C5aR N-terminal peptide. Apparent k.sub.on Apparent Apparent Antibody Antigen Name [1/Ms] k.sub.off [1/s] K.sub.D [nM] Comment MAB#2 Human 3.78E+05 1.11E-04 0.29 MAB#1 C5aR_NT 4.76E+05 9.99E-06 0.02 MAB#2 Cynomolgus 1.38E+05 1.89E-02 140 fast k.sub.off MAB#1 C5aR_NT 1.71E+05 1.08E-02 63 fast k.sub.off

Example 6: Apparent Affinity (Bivalent) Determination for MAB#1 on Full-Length C5aR Expressed on Cells by Using KinExA

[0555] Since C5aR belongs to the family of GPCRs, it is very difficult to generate recombinant full-length antigen material that can be used for SPR measurements. Therefore, Flp-In CHO cells stably expressing human C5aR or cynomolgus C5aR and KinExA measurements were performed.

Method

[0556] Apparent K.sub.D determination on C5aR-expressing cells was performed at RT with a KinExA 3200 instrument (Sapidyne Instruments). PBS (Gibco), supplemented with BSA (1 mg/mL) and 0.02% (v/v) NaN.sub.3 was used as assay buffer. MAB#1 (conc.: of 2 nM and 30 pM) was used as analyte and Flip-In CHO hC5aR_V5/His cells (final concentration 3 Mio cells/mL and 1 Mio cells/mL, 2n serial dilution) or Flip-In CHO cyC5aR_V5/His cells (final concentration 25 Mio cells/ml and 6 Mio cells/ml, 2n serial dilution) as titrant and equilibrated over night at RT in an overhead shaker. After equilibration, the samples were centrifuged and the supernatants were used for analysis. Free concentration of MAB#1 was determined using polymethylmethacrylate (PMMA) beads coated with MabSSL (GE Healthcare) and anti-human Fab2 Alexa Fluor 647 (500 ng/mL) was used for detection. The apparent K.sub.D was obtained using KinExA software and by "n-curve analysis," which fits all of the given curves to a single K.sub.D value simultaneously.

Results

[0557] Results are summarized in Table 13. MAB#1 revealed strong binding to full-length human C5aR with an apparent K.sub.D value of 130 pM. Binding to full-length cynomolgus C5aR appeared about 20.times. weaker with an apparent K.sub.D value of approx. 3 nM. Similar findings concerning binding to cynomolgus C5aR were observable before in Octet and Biacore measurements (see Example 4 and Example 5).

TABLE-US-00024 TABLE 13 Bivalent binding of MAB#1 to full-length human and cynomolgus C5aR expressed on Flp-In CHO cells Flp-In CHO Apparent K.sub.D [nM] MAB#1 human_C5aR 0.130 cyno_C5aR 2.98

Example 7: Binding of MAB#1 and MAB#2 to Full-Length C5aR Expressed on Flp-In CHO Cells and Whole-Blood Derived Neutrophils (FACS Analysis)

[0558] Cell binding to CHO Flp-In cell lines stably expressing various full-length C5aR antigens and related GPCRs as well as binding to purified human or cynomolgus neutrophils, expressing human or cynomolgus C5aR endogenously, was investigated via FACS. Cynomolgus neutrophils were obtained from whole-blood of cynomolgus monkey from three different animals (LPT Hamburg).

Methods

[0559] The C5aR-CHO Flp-In cell lines were blocked and IgGs were added either in serial dilutions or at a single (high) concentration of 300 nM. For detection of IgG binding, an R-phycoerythrin (R-PE) conjugated anti-human IgG (Fc-gamma fragment specific)2 antibody was added and fluorescence was measured using the FACS Array or Novocyte device.

[0560] Neutrophils were purified from EDTA-whole blood samples using a MACSexpress Neutrophil Isolation Cocktail from Miltenyi Biotec. Briefly, using this kit, cells are isolated using magnetic labeling and negative magnetic separation. After removal of erythrocytes, the cells were stained by adding the IgGs in a serial dilution, followed by a Alexa Fluor 647-conjugated anti-Human F(ab)2 fragment specific detection antibody. Measurements were done at the FACS Array device. FACS data were evaluated using FlowJo, entered into GraphPad Prism (v4.0) and fitted to the sigmoidal dose-response curve using non-linear regression to calculate the EC50.

Results

[0561] Results are summarized in Table 14 and FIGS. 1 and 2. FIGS. 1A and C depict binding of MAB#1, MAB#2 and RefMAB#1 to human and cynomolgus C5aR present on engineered CHO cells expressing the respective full-length receptor. Overall, very comparable binding curves on human C5aR with almost identical EC.sub.50 concentrations were determined for MAB#1, MAB#2 and RefMAB#1. Similar results were obtained on cynomolgus C5aR for MAB#1 and MAB#2. As expected, RefMAB#1 revealed no binding to cynomolgus C5aR expressed on CHO cells.

[0562] Binding to cynomolgus and human C5aR was also confirmed using purified neutrophils obtained from human or cynomolgus whole-blood (FIGS. 1B and D). Again, very comparable binding curves with almost identical EC.sub.50 values on human and cynomolgus neutrophils were observable for MAB#1. Interestingly, MAB#2 revealed overall lower signal over background levels on cynomolgus neutrophils when compared to MAB#1. This finding was not observable when binding to cynomolgus C5aR expressed on CHO cells was analyzed. Lack of binding to cynomolgus monkey neutrophils was confirmed for RefMAB#1

[0563] For rodent C5aR, no cross-reactivity to rat and mouse C5aR was expected due to the low sequence homology (66% overall identity). Indeed, neither MAB#1 nor MAB#2 showed any significant binding to rat or mouse C5aR expressed on CHO-Flp cell when tested at an IgG concentration of 300 nM (FIG. 2). In order to exclude cross-reactivity to any other member of the C5aR subfamily, binding to full-length human C5L2, ChemR23, FPR1 and C3aR expressed on CHO Flp-In cells was also determined via FACS (compared to non-transfected parental CHO Flp-In cells). As shown in FIG. 2, even at an IgG concentration of 300 nM, no significant cell binding of MAB#1 and MAB#2 was detectable to any of the C5aR-related GPCRs or to the parental CHO cells.

[0564] Taken together, both, MAB#1 and MAB#2 revealed specific binding to human and cynomolgus monkey C5aR.

TABLE-US-00025 TABLE 14 Binding of MAB#1, MAB#2 and Ref MAB#1 to full-length human or cynomolgus C5aR expressed by engineered CHO cells or purified neutrophils. Each antibody was tested in at least in two independent assay runs. Full-length protein expressed on cells MAB#1 MAB#2 RefMAB#1 human C5aR expressed on EC.sub.50 = 1.7 nM EC.sub.50 = 1.2 nM EC.sub.50 = 1.1 CHO cells nM C5aR expressed on purified EC.sub.50 = 6.0 nM ECK = 5.4 nM EC.sub.50 = 3.1 human neutrophils nM cynomolgus C5aR expressed EC.sub.50 = 1.1 nM EC.sub.50 = 1.4 nM No binding on CHO cells C5aR expressed on purified EC.sub.50 = 4.5 nM EC.sub.50 = 3.8 nM No binding cynomolgus neutrophils

Example 8: Binding of MAB#1 and MAB#2 to Full-Length Human C5aR Displayed on Virus-Like-Particles (VLPs)--Binding to Natural Variants of Human C5aR

[0565] Two natural variants of human C5aR (SEQ ID NO: 1; D/K variant and SEQ ID NO: 2; N/N variant) are reported.

[0566] To compare binding of MAB#1 and Mab#2 to the two natural variants, VLPs expressing either of the two C5aR variants were generated as described in Example 1. As negative control, VLPs expressing murine C5aR were included, since MAB#1 and Mab#2 are not cross-reactive to murine C5aR.

Method

[0567] For assessment of binding, produced VLPs were coated overnight and after a blocking step, IgG titrations were added on the next day. Binding of the IgGs to the coated antigen was detected using an Alkaline Phosphatase-conjugated anti-human IgG2 antibody and AttoPhos as substrate.

Results

[0568] Results are summarized in Table 15 and depicted in FIG. 3. Both, MAB#1 and MAB#2 revealed comparable titration curves on both natural variants of human C5aR with equal EC.sub.50 concentrations. As expected, no binding to murine C5aR was detected. Based on these data, high affinity binding of MAB#1 and MAB#2 to human C5aR can be expected in vivo, independent of the natural variant present on the respective target cells.

TABLE-US-00026 TABLE 15 Binding of MAB#1 and MAB#2 to two natural variants of human C5aR expressed on VLPs. C5aR variant MAB#1 MAB#2 huC5aR (D/K) EC.sub.50 = 0.5 nM (N = 2) EC.sub.50 = 0.5 nM (N = 2) huC5aR (N/N) EC.sub.50 = 0.5 nM (N = 2) EC.sub.50 = 0.5 nM (N = 2)

Example 9: Protein Panel Profiling (3P)

[0569] Potential unspecific off-target binding for MAB#1 was determined in the generic 3P assay.

Method:

[0570] Protein panel profiling was mainly performed as described by Frese et al. (mAbs 5:2, 279-287; March/April 2013). 32 different proteins and controls were coated on two 384-well MSD standard plates at a concentration of 1.0 .mu.g/mL at 4.degree. C. over night. The coating solution was discarded and plates were blocked with 50 .mu.L 3% (w/v) BSA in PBS for one hour at RT on a microtiter plate shaker (.about.500 rpm) followed by three washing steps with 50 .mu.L washing buffer (PBS with 0.05% (v/v) Tween 20). IgG samples were diluted to 100 nM and 10 nM in assay buffer (PBS with 0.5% (w/v) BSA, 0.05% (v/v) Tween 20). As controls, isotype control antibody MOR03207 (IgG1f_AEASS) and assay buffer were used. Samples and controls were added at 30 .mu.L/well and incubated for three hours at RT on a microtiter plate shaker. The plates were washed three times and 30 .mu.L detection antibody (ECL-labeled anti-human Fab) were added per well and incubated for one hour on a microtiter plate shaker (.about.500 rpm). After washing the MSD plate and adding 35 .mu.L/well MSD Read Buffer T with surfactant, electro-chemiluminescence signals were detected using a SECTOR Imager S600 instrument (Meso Scale Diagnostics).

[0571] For evaluation, binding signals of the antibody sample to a certain protein were divided by the respective binding signals of the reference antibody MOR03207 resulting in a binding ratio (BR). The cumulative binding ratio (CBR) of all proteins except the controls (25 in total) was then calculated: CBR up to 150 indicates an IgG without detectable unspecific binding. Values above 150 indicates an IgG with increased unspecific binding compared to the reference antibody MOR03207.

Results

[0572] Results for MAB#1 and MAB#2 are summarized in FIG. 10. In sum, no critical unspecific binding was detectable to any of the tested proteins. Very low (MAB#2) and low (MAB#1) binding to bovine transferrin was observed. Binding to bovine transferrin was confirmed in an Octet based binding assay, but no binding to rat and cynomolgus transferrin was detected (data not shown). Thus, the observed binding to bovine transferrin was regarded as non-critical.

Final Antibody Format and Safety

[0573] C5aR is expressed on various immune cells, such as leukocytes, neutrophils and lymphocytes and human IgG1 Fc-mediated depletion of such cells needs to be prevented to avoid unwanted side effects. Accordingly, the final IgG format of an anti-C5aR antibody needs to be silent in terms of its ability to induce any effector function during therapeutic intervention.

[0574] Both, MAB#1 and MAB#2 contain five amino acid substitutions in the Fc region of human IgG1f, namely L234A, L235E, G237A, A330S and P331S (hIgG1f_AEASS, numbering according EU index) to abolish antibody induced effector function. Clinical safety of this format in the context of C5aR antibody therapy has been described in the art (Wagner F et al. Annals of the Rheumatic Diseases. 2014; 73: 499. doi: 10.1136/annrheumdis-2014-eular.2156.)

[0575] The lack of ability of MAB#1 and MAB#2 to induce effector function was confirmed in various assays, such as binding studies to Fc.gamma. receptors or C1q as well as in vitro ADCC and ADCP assays as outlined below.

Example 10: Binding of MAB#1 and MAB#2 to FcRn Receptor Using Octet

Method

[0576] Apparent K.sub.D determination to immobilized neonatal Fc receptor (FcRn) from different species was performed at pH 6.0 and 7.2 at 27.degree. C. using an Octet HTX instrument (ForteBIO, Pall Life Sciences). 0.5 nm of biotinylated human, cynomolgus, mouse and rat FcRn were captured on streptavidin (SA) sensors (ForteBIO, Pall Life Sciences). Kinetic measurements were performed using 8 different concentrations of IgGs (3n serial dilution, 1000 to 0.46 nM) in Octet buffer (PBS, 0.05% (w/v) Tween-20, 0.1% (w/v) BSA) with 240 s association time and 180 s dissociation time. After each cycle the sensor was regenerated to remove bound ligand/antibody complex (2.times.30 s in HBS-EP+, pH 8.0). All sensorgrams were fitted using Data Analysis Software 10.0 (ForteBIO, Pall Life Sciences) to determine the apparent affinity and the data was fitted with a steady state model.

Results

[0577] Results are summarized in Table 16. MAB#1 and MAB#2 revealed apparent binding affinities to FcRn from different species in an expected affinity range (comparable to isotype control antibody MOR03207 IgG1f and the physiological binding behavior for binding to human FcRn could be confirmed for both IgG molecules, i.e. no binding at neutral pH (7.2) was detectable. Accordingly, the introduced 5 mutations into the Fc region of the antibodies did not adversely affected human FcRn binding.

TABLE-US-00027 TABLE 16 Binding of MAB#1 and MAB#2 to FcRn via the Fc region at pH 6.0 and 7.2. K.sub.D [nM] K.sub.D [nM] Antibody Antigen pH 6.0 pH 7.2 MAB#2 Human FcRn 36* no binding MAB#1 9.0* no binding MAB#2 Cynomolgus 30* no binding MAB#1 FcRn 6.8* 560** MAB#2 Murine FcRn 7.3 130 MAB#1 4.3 39 MAB#2 Rat FcRn 10 300 MAB#1 5.7 110 *Deviation from fit model (pH 6.0) **Slight binding at pH 7.2 *** Values formatted in italics are mainly intended for ranking purposes

Example 11: Binding of MAB#1 and MAB#2 to Human Fc.gamma. Receptors Using Octet

Method

[0578] K.sub.D determination via IgG capture setup was performed at 27.degree. C. using Octet (ForteBIO, Pall Life Sciences). 2.0 nm of IgGs diluted in Octet assay buffer (PBS, 0.05% (v/v) Tween-20, 0.1% (w/v) BSA) were captured on Protein A sensors (ForteBIO, Pall Life Sciences). Kinetic measurements were performed using 7 concentrations of Fc gamma receptors (2n serial dilution) in assay buffer. After each cycle the sensors were regenerated to remove bound ligand/antibody complex (2.times.30 s in 10 mM Gly/HCl, pH 1.5). All sensorgrams were fitted using Data Analysis Software 10.0, (ForteBIO, Pall Life Sciences) to determine k.sub.on and k.sub.off rate constants, which were used to calculate K.sub.D. The raw data was fitted with a 1:1 binding model, with parameter R.sub.max set to local.

Results

[0579] Results are summarized in Table 17. No or only very weak binding of MAB#1 and MAB#2 to any of the tested Fc.gamma. receptors could be detected and confirmed that the introduced mutations into the human IgG1 Fc region are effective in abolishing Fc.gamma. receptor binding.

TABLE-US-00028 TABLE 17 Binding of MAB#1 and MAB#2 to human Fc.gamma. receptors via Fc-region Fc.gamma.R MAB#2 Mab#1 hu_Fc.gamma.RI no binding no binding hu_Fc.gamma.RIIa (131H) no binding no binding hu_Fc.gamma.RIIa(131R) very slight binding very slight binding only at highest only at highest antibody concentration antibody concentration observed observed hu_Fc.gamma.RIIIa (158F) no binding no binding hu_Fc.gamma.RIIIa (158V) no binding no binding hu_Fc.gamma.RIIb no binding no binding

Example 12: Binding of MAB#1 and MAB#2 to C1q Using Octet

Method

[0580] Apparent (bivalent) K.sub.D determination via IgG capture setup was performed at 27.degree. C. using Octet (Fortebio Pall Life Sciences). 2.0 nm of IgGs diluted in Octet buffer were captured onto anti-hu Fab CH1 kappa/lambda (BAC) immobilised onto streptavidin (SA) sensors (ForteBIO, Pall Life Sciences). Kinetic measurements were done using 8 concentrations of C1q (3n serial dilution, 500 to 0.69 nM) in Octet assay buffer (see above) with 240 s association time and 240 s dissociation time. After each cycle the sensors were regenerated to remove bound ligand/antibody complex (2.times.50 mM NaOH, 1.times.10 mM Gly/HCl, pH 1.5, for 30 s each). All sensorgrams were fitted using Data Analysis Software 9 (ForteBIO, Pall Life Sciences) to determine the apparent affinity. The data was fitted with a steady state model.

Results

[0581] Results are summarized in Table 18. As expected, no binding of MAB#1 and MAB#2 to C1q (isolated from pooled human plasma) was observable and confirmed that the introduced mutations into the human IgG1 Fc region are effective in abolishing C1q binding.

TABLE-US-00029 TABLE 18 Binding of MAB#1 and MAB#2 to human C1q. Antigen MAB#2 MAB#1 Human C1q no binding no binding

Example 13: ADCC and ADCP In Vitro Activity of MAB#1

Methods

[0582] ADCC and ADCP activity for MAB#1 was tested using the Promega ADCC and ADCP Reporter Bioassays according to the manufacturer's instructions (Cat #G7017 and Cat #G988A, respectively). The kits employs engineered Jurkat cells as effector cells. The cells either stably express the Fc.gamma.RIIIa receptor, V158 (high affinity) variant for ADCC and Fc.gamma.RIIa_H receptors for ADCP and an NFAT response element driving expression of firefly luciferase. As target cells, CHO Flp-In cells expressing human C5aR were used. Binding of the effector cells to the target through the antibody bridge (e.g. through MAB#1 or MAB#2) initiates a cascade of events in the NFAT pathway, resulting in the expression of the firefly luciferase protein. The enzymatic reaction produces luminescence, which is proportional to the luciferase concentration, directly correlating to ADCC or ADCP activity. ADCC or ADCP activity was analyzed for MAB#1 by plotting the average signal to background values.

[0583] Since for MAB#1, a wild-type human IgG1f version was not available, a wild-type IgG1f as well as a Fc-silent human IgG1f_AEASS version of an in-house human anti-C5aR control antibody was included as positive control. In addition, a Fc-silent (hIgG1_AEASS) and wild-type version (hIgG1f) version of the isotype control antibody MOR03207 was included as negative control.

Results

[0584] Results are summarized in Table 19 and depicted in FIGS. 8A (ADCP) and B (ADCC) for an IgG concentration of 10 .mu.g/ml. MAB#1 did not induce Fc.gamma.RIIIa or Fc.gamma.RIIa_H activation of the NFAT pathway in engineered effector cells in the presence of C5aR overexpressing CHO cells, similar to the Fc-silent version of the anti-C5aR control antibody and the Fc-silent version of MOR03207. The wild-type (non-silent) version of the C5aR specific control antibody clearly induced luciferase production in engineered Jurkat cells in the presence of C5aR expressing CHO cells.

[0585] In sum, the experiment clearly confirmed that the introduced mutations into the wild-type human IgG1 Fc region are efficient in preventing ADCC and ADCP activity.

TABLE-US-00030 TABLE 19 Overview of the conducted in vitro assays to confirm that MAB#1 (hIgG1f_AEASS) does not mediate effector function. Criteria Assay Results for MAB#1 No ADCC ADCC Reporter No Fc.gamma.RIIIa_V activation of NFAT Bioassay pathway in engineered effector cells detectable No ADCP ADCP Reporter No Fc.gamma.RIIa_H activation of NFAT Bioassay pathway in engineered effector cells detectable No CDC huC1q Binding No binding to human C1q detectable (Octet) No binding Fc.gamma.R Binding No binding to the to Fc.gamma. (Octet) Fc.gamma.Rs. Highest background Receptors (close to slight binding) on hFc.gamma.RIIa_R Physiological FcRn Binding Physiological binding FcRn (Octet) with apparent binding K.sub.D in the expected range

Functional Characterization of MAB#1 and MAB#2

[0586] The neutralizing activities of MAB#1 and MAB#2 were analyzed in different in vitro assays which monitor C5a induced activation of C5aR.

[0587] As elevated levels of C5a has been described under pathophysiological conditions, the capability of a C5aR antagonistic antibody to neutralize high concentrations of C5a, which may be present locally at the disease site, is expected to provide a beneficial therapeutic effect in vivo. Accordingly, the in vitro experiments were also set-up to reflect such in vivo pathological conditions.

Example 14: PathHunter.RTM. .beta.-Arrestin Assay (DiscoveRx)

Methods:

[0588] The PathHunter.RTM. .beta.-Arrestin assay from DiscoveRx was performed according to the manufacturer's instructions. In brief, human C5a induced human C5aR activity was measured by the detection of the interaction of .beta.-arrestin with the activated C5aR using .beta.-galactosidase enzyme fragment complementation.

[0589] .beta.-arrestin recruitment was induced using recombinant human C5a and enzyme activity was measured using chemiluminescent detection reagents from DiscoverRx. Chinese hamster ovarian cells (CHO) cells, expressing the engineered version of human C5aR were seeded overnight and serial dilutions of human C5a (R&D Systems) were added and incubated at 37.degree. C. and 5% CO2 for 1.5 h (generating the titration curves in absence of the antagonist). In parallel, titration curves of human C5a were determined in presence of the C5aR specific antibodies (fixed IgG conc. of 50 nM). For doing so, IgGs were added to the cells before stimulation with human C5a and incubated for 1 h at 37.degree. C. and 5% CO.sub.2. Results were expressed as relative luminescence units. Titration curves for human C5a in the presence and absence of the antagonistic IgGs were generated via GraphPad Prism.

[0590] For the .beta.-arrestin assay, the shifts in the dose-response curves for human C5a to higher doses (i.e. horizontally to the right on the dose axis) were compared for MAB#1, MAB#2 and RefMAB#1 (FIG. 4A). Additionally, the % inhibition at three increasing C5a concentrations (1.2 nM, 11 nM and 100 nM) was calculated and compared (FIG. 4B).

Results

[0591] The results from the .beta.-arrestin assays are depicted in FIGS. 4A and 4B and summarized in Table 20 and Table 21. In the absence of antibody, a dose-response curve for human C5a with an average EC.sub.50, concentration of 2.9 nM was obtained (FIG. 4A).

[0592] Adding MAB#1 or MAB#2 to human C5a at a final IgG concentration of 50 nM resulted in a significant shift in the dose-response curve of more than 12-fold to higher doses of human C5a. Being more precisely, in the presence of 50 nM MAB#1 or MAB#2, a dose-response curve for human C5a with an average EC.sub.50 concentration of 37 nM was obtained (FIG. 4A, Table 20). In other word, the presence of MAB#1 or MAB#2 significantly reduces the ability of human C5 to induce C5aR activation or, alternatively, in the presence of MAB#1 or MAB#2, human C5a needs to be added in an at least 12-fold higher concentration in order to induce the same C5aR activity when compared to its activity in the absence of antibody.

[0593] This observation was also reflected when the % inhibition at three increasing C5a concentrations was calculated (FIG. 4B). At 1.2 nM human C5a, all three tested antibodies, MAB#1, MAB#2 and RefMAB#1, revealed a comparable inhibition of C5a induced C5aR activity of more than 80% at an IgG concentration of 50 nM. If however, an about 10-fold higher concentration of human C5a (11 nM) was used for activation of C5aR, RefMAB#1 was only able to neutralize of about 50% of C5a induced C5aR activity. This is in strong contrast to MAB#1 and MAB#2, which were still able to neutralize up to 80% of C5a induced C5aR activity.

[0594] This effect was even more pronounced when a 100-fold higher concentration of human C5a (100 nM) for activation of C5aR was used. Here, almost no neutralizing activity was detectable for RefMAB#1, whereas MAB#1 and MAB#2 were still able to neutralize of about 40% and 30%, respectively, of the human C5a induced C5aR activity.

[0595] Accordingly, both, MAB#1 and MAB#2 are efficient in neutralizing pathophysiological C5a concentrations in vitro and are significant more potent compared to RefMAB#1.

TABLE-US-00031 TABLE 20 In vitro .beta.-arrestin assay (n = 3): Inhibitory activity of C5aR specific antibodies on C5a-induced C5aR activity measured by the detection of the interaction of .beta.-arrestin with activated C5aR using .beta.-galactosidase enzyme fragment complementation. Activity of human C5a x fold reduced EC.sub.50 (nM) in absence or activity of C5a presence of in presence anti-C5aR antibody of anti C5aR antibody hC5a 2.9 -- hC5a + MAB#1 (50 nM) 37 12.8 hC5a + MAB#2 (50 nM) 37 12.8 hC5a + RefMAB#1 10 3.4 (50 nM)

TABLE-US-00032 TABLE 21 In vitro .beta.-arrestin assay (n = 3): Inhibitory activity of C5aR specific antibodies on C5a-induced C5aR activity calculated for 3 concentrations of C5a and a fixed IgG concentration of 50 nM and measured by the detection of the interaction of .beta.-arrestin with activated C5aR using .beta.-galactosidase enzyme fragment complementation. MAB#2 MAB#1 RefMab#1 [%] inhibition at 1.2 nM C5a 90.5 91.9 84.2 and 50 nM IgG [%] inhibition at 11 nM C5a 75.2 79.9 38.9 and 50 nM IgG [%] inhibition at 100 nM C5a 31.2 41.8 6.4 and 50 nM IgG

Example 15: Inhibition of Neutrophil and Monocyte Activation by MAB#1 and MAB#2-CD11b Assay

[0596] The neutralization potency of MAB#1 and MAB#2 was furthermore determined in a functional CD11b whole blood assay representing a more physiological set up. CD11b combines with CD18 to form the integrin Mac-1 complex, which serves as a multi-ligand receptor. CD11b is constitutively expressed on the surface of >50% peripheral blood leukocytes; upon leukocyte activation, its expression is up regulated through the fusion of CD11b containing secretory granules into the cell membrane. CD11b expression is thus widely used as a marker of leukocyte activation both in vivo and in vitro.

[0597] C5a, as a potent activator of human neutrophils and monocytes, induces up-regulation of surface antigen CD11b. Thus, the ability of MAB#1 and MAB#2 to prevent C5a-induced activation of granulocytes and monocytes was investigated by assessing the CD11b levels in whole-blood derived granulocytes and monocytes. The experiments were basically performed as described in US patent application US2013/0295116 (NOVO NORDISK).

Method

[0598] In a first assays set-up, whole heparinized blood was mixed with IgG (in serial dilutions) and incubated for 20 min at 37.degree. C., 5% CO.sub.2. Human C5a was added at a standard concentration of 15 nM and incubated for 20 min at 37.degree. C., 5% CO.sub.2. Anti-CD11b-PE or isotype control antibody MOR03207 was added and the plate was incubated for 20 min. at 37.degree. C., 5% CO.sub.2. Finally, a red blood cell lysing buffer was added and incubated at room temperature for 15 min in the dark, the cells were washed and again re-suspended in lysing buffer.

[0599] In a second experimental set-up, human C5a was added at a more clinical relevant pathophysiological concentration of 150 nM without modifying the remaining assays set-up as described above.

[0600] To investigate the receptor residence time of MAB#1, the assay was further adapted by prolonging the incubating time of the heparinized whole blood with IgG from 20 min to 300 min.

[0601] Fluorescence was measured using the FACS Array or Novocyte device. Samples were gated to exclude dead cells and debris. Monocytes and granulocytes were identified according to their FSC and SSC profiles and gated. The median fluorescence intensity (MFI) of the gated granulocytes and/or monocytes in the CD11b-PE channel (Yellow-A) was calculated. Results were expressed as a percentage of inhibition (% Inhibition). Maximum CD11b expression (MFI.sub.max) was the average MFI of the cells, incubated with C5a but without IgG. The minimum (background) CD11b expression (MFI.sub.Min) was the average MFI of the cells incubated without C5a and without IgG. The formula used to calculate % inhibition for each samples was:

% Inhibition=100-(((MFI.sub.sample-MFI.sub.Min))/((MFI.sub.Max-MFI.sub.M- in)).times.100)

[0602] Data was evaluated using FlowJo, entered into GraphPad Prism (v4.0) and fitted to the sigmoidal dose-response curve using non-linear regression to calculate the IC.sub.50.

Results for MAB#1 Using 15 nM C5a Ligand

[0603] Results for MAB#1 from the CD11b whole blood assays are summarized in Table 22. For both gated cell populations (monocytes and granulocytes), IC.sub.50 values in the single-digit nM range were determined with maximum inhibition of almost 88% for granulocytes and 83% for monocytes.

TABLE-US-00033 TABLE 22 Ability of MAB#1 to prevent C5a-mediated activation of granulocytes and monocytes. Overview maximum inhibition and IC.sub.50 values was determined in the CD11b assay using 15 nM C5a (n = 3). Cell type MAB#1 - Inhibition of CD11b expression CD11b max % inhibition (at 600 nM IgG) 87.2 .+-. 1.7 (N = 3) Granulocytes IC.sub.50 (nM) 9.8 .+-. 0.6 (N = 3) CD11b max % inhibition (at 600 nM IgG) 83.3 .+-. 3.6 (N = 3) Monocytes IC.sub.50 (nM) 4.2 .+-. 0.8 (N = 3)

Results for MAB#1, MAB#2 and RefMAB#1 Using 15 nM Vs. 150 nM C5a Ligand

[0604] Besides adding 15 nM of human C5a for stimulation of granulocytes (as described above), a ten-fold higher concentration (150 nM) of ligand was used to simulate a more pathophysiological ligand concentration.

[0605] Again, MAB#1, MAB#2 and RefMAB#1 were dose-titrated and inhibition curves were generated using GraphPad Prism via the nonlinear regression function. The results are quantitatively expressed as "concentration of antagonist needed in order to induce 50% of inhibition".

[0606] The results from this CD11b whole blood assay are shown in FIGS. 5A and B.

[0607] FIG. 5A depicts the neutralizing activity of MAB#1, MAB#2 and RefMAB#1 at 15 nM C5a and FIG. 5B depicts the respective neutralizing activity at 150 nM C5a. As a quantitative read-out, the IgG concentration for reaching 50% inhibition of CD11b upregulation was calculated and the results are indicated below the x-axis in both Figures.

[0608] Overall, similar observations as for the .beta.-arrestin assay (Example 14) were made when using increasing C5a concentrations in the CD11b whole blood assay gated on granulocytes as primary target cells. In order to block the effects of 15 nM human C5a to 50%, an IgG concentration of 12 nM for MAB#1 and RefMAB#1 was needed, whereas for MAB#2 an IgG concentration of 17 nM was sufficient. However, in order to block 50% of CD11b upregulation induced by a 10 fold higher C5a concentration (150 nM) significant different amounts of the antagonists were required. While for MAB#1, an IgG concentration of 42 nM were now sufficient to inhibit 50% of the C5a induced activation of C5aR, a 7-fold higher concentration of RefMAB#1 (291 nM) was required to reach the same blocking effect. For MAB#2 an IgG concentration of 114 nM was sufficient.

[0609] Accordingly, the same ranking of the 3 IgGs in terms of potency as already seen in the b-arrestin assay (MAB#1>MAB#2>RefMAB#1) could be done and both, MAB#1 and MAB#2 appeared very efficient in neutralizing pathophysiological C5a concentrations in vitro and were significant more potent compared to RefMAB#1.

Results for MAB#1--Influence on Receptor Residence Time

[0610] Seow and colleagues (Seow V et al., Sci Rep. 2016; 6: 24575) reported that the generation of C5a is localized at the cell membrane and can be profoundly high for brief but repeated periods. Therefore, it was suggested that an antagonist of C5aR with long residence time could be advantageous in systems with rapid and transient signaling. It was also concluded that an increased receptor residence time measured in vitro could also translate to the duration of action and degree of efficacy in vivo.

[0611] To compare the receptor residence time of the two antibodies in a similar set up as published by Seow and co-workers, log dose inhibition curves for 20 minutes and 300 minutes of IgG incubation with granulocytes and monocytes present in whole blood was evaluated as described above.

[0612] The results from this experimental set-up of the CD11b whole blood assay are shown in FIG. 6A-D. Surprisingly, a significant increase in the neutralizing activity over a prolonged period of incubation time was observable of MAB#1. As shown in FIGS. 6A and C, for both, granulocytes and monocytes populations, the calculated IC.sub.50 concentration for MAB#1 decreased over time of about 6-fold (IC.sub.50 values are summarized in Table 23). For RefMAB#1, no shift or decrease in the inhibition curves was observable after 300 minutes (FIGS. 6B and D).

TABLE-US-00034 TABLE 23 CD11b assay after 20 minutes or 300 minutes of IgG incubation with whole-blood at 15 nM C5a. Provided are IC.sub.50 values and x-fold potency improvements (20 min. vs 300 min.) for granulocytes (upper panel) and monocytes (lower panel) MAB#1 RefMAB#1 CD11b x-fold x-fold Granulocytes IC50 (in nM) improvement IC50 (in nM) improvement 20 minutes 11.0 .+-. 1.0 4.8 10.2 .+-. 1.9 1.6 (N = 2) (N = 2) 300 minutes 2.3 .+-. 0.0 6.5 .+-. 0.5 (N = 2) (N = 2) MAB#1 RefMAB#1 CD11b x-fold x-fold Monocytes IC50 (in nM) improvement IC50 (in nM) improvement 20 minutes 4.0 .+-. 1.0 5.7 1.8 .+-. 0.5 0.3 (N = 2) (N = 2) 300 minutes 0.7 .+-. 0.2 5.3 .+-. 1.0 (N = 2) (N = 2)

[0613] In sum, the combination of both, efficient neutralization of pathophysiological C5a levels and an increased potency over time is expected to be beneficial in vivo.

Example 16: C5a Induced Migration of Neutrophils

[0614] C5a induced chemotaxis of purified neutrophils was analyzed. The experiment was basically performed as described in US patent application US2013/0295116.

Methods

[0615] Neutrophils were isolated and purified from whole blood of three different human donors using the MACSxpress Whole Blood Neutrophil Isolation Kit, human (Miltenyi Biotec Cat #130-104-434 and the MACSxpress Erythrocyte Depletion Kit, human (Miltenyi Biotec, CAT #130-098-196).

[0616] The potency of the IgGs to inhibit hC5a dependent neutrophil migration was analyzed by the Boyden chamber technique using Fluor.RTM. Blok.RTM. 3.0 .mu.M pore size 96-well plates. The membrane of the Boyden chamber pore was coated with 1 mg/mL human fibrinogen for 2 hrs at 37.degree. C. After washing, the membranes were blocked with a solution containing 2% bovine serum albumin (BSA) for 1 hr at 37.degree. C. Purified neutrophils were then stained with calcein and 10.sup.5 stained cells added to the upper compartment in the Boyden chamber with and without the antagonistic IgGs (100 and 600 nM). hC5a (R&D Systems, 10 nM) was applied to the lower compartment in the Boyden chamber. The plate was measured at 485/538 nm at 37.degree. C. every 5 min for 60 min in a plate reader (Tecan M1000Pro). The ability of neutrophils to migrate to the lower chamber is determined measuring the calcein-stained neutrophils passing through the fluoroblok membrane. The results are expressed as kinetic migration curves (5-60 min) as well as percentage inhibition calculated at selected time points (15, 25 and 35 min).

The formula used to calculate % inhibition was

% Inhibition=100-(((RFU.sub.sample-RFU.sub.Min))/((RFU.sub.Max-RFU.sub.M- in)).times.100)

Results

[0617] The average percentage inhibition of neutrophil migration at selected time points at two tested IgG concentrations was calculated from 3 independent experiments using neutrophils from three different human donors.

[0618] As depicted in FIG. 7, after 15 min of migration, almost complete inhibition of C5a-induced neutrophil migration was reached for MAB#1. After 35 min of migration and the highest IgG concentration tested (600 nM), MAB#1 still revealed >50% inhibition. The negative isotype control antibody MOR03207 showed no inhibitory effect on neutrophil migration.

Example 17: C5a Induced Release of Cytokines by Macrophages

[0619] Macrophages release pro- or anti-inflammatory cytokines upon activation depending on their polarization. Hence, it was evaluated whether blockade of the C5a/C5aR interaction with MAB#1 affects the C5a-induced release of cytokines by M1 and M2 macrophages. A cytokine ELISA was performed to detect the production of anti-inflammatory IL-10 by M2 macrophages and pro-inflammatory IL-12 by M1 macrophages.

Methods

[0620] Briefly, peripheral blood mononuclear cells (PBMCs) were prepared from whole blood of healthy human donors by density-gradient centrifugation with Biocoll separating solution and SepMate tubes (Stemcell). Monocytes were isolated from PBMCs using a CD14+ selection kit (Miltenyi Biotech) and cultured in RPMI supplemented with 10% FCS and 1.times.GlutaMax in 96 well plates. Overnight-plated monocytes were polarized for 24 hours at 37.degree. C. to M1 macrophages using LPS (20 ng/ml) and IFN.gamma. (50 ng/ml) and pre-incubated with MAB#1 (30 nM) for 30 minutes followed by addition of C5a (15 nM) for another 24 hours. M1 macrophages stimulated with C5a in absence of MAB#1 and untreated controls were included. After 24 hours, cell supernatants were analyzed by an IL-12/IL23 DuoSet ELISA (R&D Systems) according to manufacturer's instructions. For generation of M2 macrophages, monocytes were pre-differentiated into macrophages by culture for 5 days in RPMI/10% FCS supplemented with M-CSF and addition of M-CSF, IL-4, IL-13 and IL-6 (40 ng/ml each) at day 5. C5a (15 nM)+/-MAB#1 (30 nM) were added daily from day 5 until 9. On day 9, cell supernatants were analyzed by an IL-10 DuoSet ELISA (R&D Systems) according to manufacturer's instructions.

Results

[0621] While incubation with C5a lead to decreased IL-12 production of M1 macrophages, no reduction in IL-12 levels could be observed after pre-treatment with MAB#1 in comparison to the untreated control. On the other hand, treatment with MAB#1 inhibited C5a-induced IL-10 production in M2 macrophages (see FIG. 11)

[0622] In conclusion, MAB#1 efficiently inhibited C5a-induced production of anti-inflammatory IL-10 by M2 macrophages and restored the production of pro-inflammatory IL-12 by M1 macrophages thereby demonstrating the mode of action of MAB#1 in vitro.

Pharmacokinetics

Example 18: Pharmacokinetics

[0623] The pharmacokinetic profile of MAB#1 was assessed in male Han-Wistar rats (n=3 animals) after a single intravenous (i.v.) administration of 10 mg/kg IgG.

Methods

[0624] Plasma samples were collected from each animal via retro-orbital sinus or mandibular vein puncture at the following time-points: Predose, 0.083, 1, 3, 8, 24, 48, 72, 96, 168, 240, 336 and 504 hours post administration.

[0625] Free, bioactive MAB#1 concentrations in rat plasma were determined using a MSD-based ligand-binding assay. Briefly, a biotinylated N-terminal human C5aR peptide was coated on the surface of a 96-well Streptavidin-MSD plate. The bound analyte was detected using a drug-specific anti-idiotypic ECL-labelled antibody. Pharmacokinetic properties of MAB#1 were evaluated using non-compartmental data analysis (NCA) based on free drug concentrations in plasma.

Results

[0626] The mean plasma concentrations over time is depicted in FIG. 9. The mean maximum plasma concentrations of MAB#1 after a single i.v. administration were observed at 5 minutes (i.e. 0.083 hours) after administration (T.sub.max) in all three animals (i.e. first sampling time point post administration). The mean volume of distribution (Vz) of 106 mL/kg was between plasma volume and extracellular volume (Davies et al., 1993). The mean terminal elimination half-life following i.v. administration was determined at 9.0 days, the mean total clearance was determined at 0.341 mL/h/kg.

[0627] Overall, MAB#1 demonstrated a typical pharmacokinetic profile of a human IgG1 antibody in rat plasma with no cross-reactivity to the rodent C5aR. No signs of anti-drug-antibody (ADA)-mediated clearance could be detected.

Sequence CWU 1

1

961350PRTHomo sapiens 1Met Asp Ser Phe Asn Tyr Thr Thr Pro Asp Tyr Gly His Tyr Asp Asp1 5 10 15Lys Asp Thr Leu Asp Leu Asn Thr Pro Val Asp Lys Thr Ser Asn Thr 20 25 30Leu Arg Val Pro Asp Ile Leu Ala Leu Val Ile Phe Ala Val Val Phe 35 40 45Leu Val Gly Val Leu Gly Asn Ala Leu Val Val Trp Val Thr Ala Phe 50 55 60Glu Ala Lys Arg Thr Ile Asn Ala Ile Trp Phe Leu Asn Leu Ala Val65 70 75 80Ala Asp Phe Leu Ser Cys Leu Ala Leu Pro Ile Leu Phe Thr Ser Ile 85 90 95Val Gln His His His Trp Pro Phe Gly Gly Ala Ala Cys Ser Ile Leu 100 105 110Pro Ser Leu Ile Leu Leu Asn Met Tyr Ala Ser Ile Leu Leu Leu Ala 115 120 125Thr Ile Ser Ala Asp Arg Phe Leu Leu Val Phe Lys Pro Ile Trp Cys 130 135 140Gln Asn Phe Arg Gly Ala Gly Leu Ala Trp Ile Ala Cys Ala Val Ala145 150 155 160Trp Gly Leu Ala Leu Leu Leu Thr Ile Pro Ser Phe Leu Tyr Arg Val 165 170 175Val Arg Glu Glu Tyr Phe Pro Pro Lys Val Leu Cys Gly Val Asp Tyr 180 185 190Ser His Asp Lys Arg Arg Glu Arg Ala Val Ala Ile Val Arg Leu Val 195 200 205Leu Gly Phe Leu Trp Pro Leu Leu Thr Leu Thr Ile Cys Tyr Thr Phe 210 215 220Ile Leu Leu Arg Thr Trp Ser Arg Arg Ala Thr Arg Ser Thr Lys Thr225 230 235 240Leu Lys Val Val Val Ala Val Val Ala Ser Phe Phe Ile Phe Trp Leu 245 250 255Pro Tyr Gln Val Thr Gly Ile Met Met Ser Phe Leu Glu Pro Ser Ser 260 265 270Pro Thr Phe Leu Leu Leu Lys Lys Leu Asp Ser Leu Cys Val Ser Phe 275 280 285Ala Tyr Ile Asn Cys Cys Ile Asn Pro Ile Ile Tyr Val Val Ala Gly 290 295 300Gln Gly Phe Gln Gly Arg Leu Arg Lys Ser Leu Pro Ser Leu Leu Arg305 310 315 320Asn Val Leu Thr Glu Glu Ser Val Val Arg Glu Ser Lys Ser Phe Thr 325 330 335Arg Ser Thr Val Asp Thr Met Ala Gln Lys Thr Gln Ala Val 340 345 3502350PRTHomo sapiens 2Met Asn Ser Phe Asn Tyr Thr Thr Pro Asp Tyr Gly His Tyr Asp Asp1 5 10 15Lys Asp Thr Leu Asp Leu Asn Thr Pro Val Asp Lys Thr Ser Asn Thr 20 25 30Leu Arg Val Pro Asp Ile Leu Ala Leu Val Ile Phe Ala Val Val Phe 35 40 45Leu Val Gly Val Leu Gly Asn Ala Leu Val Val Trp Val Thr Ala Phe 50 55 60Glu Ala Lys Arg Thr Ile Asn Ala Ile Trp Phe Leu Asn Leu Ala Val65 70 75 80Ala Asp Phe Leu Ser Cys Leu Ala Leu Pro Ile Leu Phe Thr Ser Ile 85 90 95Val Gln His His His Trp Pro Phe Gly Gly Ala Ala Cys Ser Ile Leu 100 105 110Pro Ser Leu Ile Leu Leu Asn Met Tyr Ala Ser Ile Leu Leu Leu Ala 115 120 125Thr Ile Ser Ala Asp Arg Phe Leu Leu Val Phe Lys Pro Ile Trp Cys 130 135 140Gln Asn Phe Arg Gly Ala Gly Leu Ala Trp Ile Ala Cys Ala Val Ala145 150 155 160Trp Gly Leu Ala Leu Leu Leu Thr Ile Pro Ser Phe Leu Tyr Arg Val 165 170 175Val Arg Glu Glu Tyr Phe Pro Pro Lys Val Leu Cys Gly Val Asp Tyr 180 185 190Ser His Asp Lys Arg Arg Glu Arg Ala Val Ala Ile Val Arg Leu Val 195 200 205Leu Gly Phe Leu Trp Pro Leu Leu Thr Leu Thr Ile Cys Tyr Thr Phe 210 215 220Ile Leu Leu Arg Thr Trp Ser Arg Arg Ala Thr Arg Ser Thr Lys Thr225 230 235 240Leu Lys Val Val Val Ala Val Val Ala Ser Phe Phe Ile Phe Trp Leu 245 250 255Pro Tyr Gln Val Thr Gly Ile Met Met Ser Phe Leu Glu Pro Ser Ser 260 265 270Pro Thr Phe Leu Leu Leu Asn Lys Leu Asp Ser Leu Cys Val Ser Phe 275 280 285Ala Tyr Ile Asn Cys Cys Ile Asn Pro Ile Ile Tyr Val Val Ala Gly 290 295 300Gln Gly Phe Gln Gly Arg Leu Arg Lys Ser Leu Pro Ser Leu Leu Arg305 310 315 320Asn Val Leu Thr Glu Glu Ser Val Val Arg Glu Ser Lys Ser Phe Thr 325 330 335Arg Ser Thr Val Asp Thr Met Ala Gln Lys Thr Gln Ala Val 340 345 3503350PRTMacaca fascicularis 3Met Asp Pro Phe Ser Ser Thr Thr Leu Asp Tyr Glu His Tyr Asp Gly1 5 10 15Lys Asn Val Leu Asp Ser Asp Thr Pro Val Asp Lys Thr Ser Asn Thr 20 25 30Leu Arg Val Pro Asp Ile Leu Ala Leu Val Val Phe Ala Val Val Phe 35 40 45Leu Val Gly Val Leu Gly Asn Ala Leu Val Val Trp Val Thr Ala Phe 50 55 60Glu Val Lys Arg Thr Ile Asn Ala Ile Trp Phe Leu Asn Leu Ala Val65 70 75 80Ala Asp Phe Leu Ser Cys Leu Ala Leu Pro Ile Leu Phe Thr Ser Ile 85 90 95Val Gln His His His Trp Pro Phe Gly Gly Thr Ala Cys Arg Ile Leu 100 105 110Pro Ser Leu Ile Leu Leu Asn Met Tyr Ala Ser Ile Leu Leu Leu Ala 115 120 125Thr Ile Ser Ala Asp Arg Phe Leu Leu Val Phe Asn Pro Ile Trp Cys 130 135 140Gln Asn Phe Arg Gly Ala Gly Leu Ala Trp Ile Ala Cys Ala Val Ala145 150 155 160Trp Gly Leu Ala Leu Leu Leu Thr Ile Pro Ser Phe Leu Tyr Arg Ala 165 170 175Val Arg Gln Glu Glu Tyr Ser Pro Lys Val Leu Cys Gly Val Asp Tyr 180 185 190Asn Asn Asp Thr Arg Arg Glu Arg Ala Val Ala Ile Val Arg Leu Val 195 200 205Leu Gly Phe Leu Trp Pro Leu Leu Thr Leu Met Ile Cys Tyr Thr Phe 210 215 220Leu Leu Leu Arg Thr Trp Ser Arg Arg Ala Thr Arg Ser Thr Lys Thr225 230 235 240Leu Lys Val Val Val Ala Val Val Ala Ser Phe Phe Ile Phe Trp Leu 245 250 255Pro Tyr Gln Val Thr Gly Thr Met Met Ser Phe Leu Arg Pro Ser Ser 260 265 270Pro Thr Tyr Leu Gln Leu Lys Lys Leu Asp Ser Leu Ser Ile Ser Phe 275 280 285Ala Tyr Ile Asn Cys Cys Ile Asn Pro Val Ile Tyr Val Val Ala Gly 290 295 300Gln Gly Phe Gln Gly Arg Leu Arg Lys Ser Leu Pro Ser Leu Leu Arg305 310 315 320Asn Val Leu Thr Glu Glu Ser Val Val Arg Glu Ser Lys Ser Phe Thr 325 330 335Arg Ser Thr Val Asp Thr Met Thr Glu Lys Thr Gln Ala Val 340 345 3504351PRTMus musculus 4Met Asp Pro Ile Asp Asn Ser Ser Phe Glu Ile Asn Tyr Asp His Tyr1 5 10 15Gly Thr Met Ala Pro Asn Ile Pro Ala Asp Gly Ile His Leu Pro Lys 20 25 30Arg Gln Pro Gly Asp Val Ala Ala Leu Ile Ile Tyr Ser Val Val Phe 35 40 45Leu Val Gly Val Pro Gly Asn Ala Leu Val Val Trp Val Thr Ala Phe 50 55 60Glu Ala Arg Arg Ala Val Asn Ala Ile Trp Phe Leu Asn Leu Ala Val65 70 75 80Ala Asp Leu Leu Ser Cys Leu Ala Leu Pro Val Leu Phe Thr Thr Val 85 90 95Leu Asn His Asn Tyr Trp Tyr Phe Asp Ala Thr Ala Cys Ile Val Leu 100 105 110Pro Ser Leu Ile Leu Leu Asn Met Tyr Ala Ser Ile Leu Leu Leu Ala 115 120 125Thr Ile Ser Ala Asp Arg Phe Leu Leu Val Phe Lys Pro Ile Trp Cys 130 135 140Gln Lys Val Arg Gly Thr Gly Leu Ala Trp Met Ala Cys Gly Val Ala145 150 155 160Trp Val Leu Ala Leu Leu Leu Thr Ile Pro Ser Phe Val Tyr Arg Glu 165 170 175Ala Tyr Lys Asp Phe Tyr Ser Glu His Thr Val Cys Gly Ile Asn Tyr 180 185 190Gly Gly Gly Ser Phe Pro Lys Glu Lys Ala Val Ala Ile Leu Arg Leu 195 200 205Met Val Gly Phe Val Leu Pro Leu Leu Thr Leu Asn Ile Cys Tyr Thr 210 215 220Phe Leu Leu Leu Arg Thr Trp Ser Arg Lys Ala Thr Arg Ser Thr Lys225 230 235 240Thr Leu Lys Val Val Met Ala Val Val Ile Cys Phe Phe Ile Phe Trp 245 250 255Leu Pro Tyr Gln Val Thr Gly Val Met Ile Ala Trp Leu Pro Pro Ser 260 265 270Ser Pro Thr Leu Lys Arg Val Glu Lys Leu Asn Ser Leu Cys Val Ser 275 280 285Leu Ala Tyr Ile Asn Cys Cys Val Asn Pro Ile Ile Tyr Val Met Ala 290 295 300Gly Gln Gly Phe His Gly Arg Leu Leu Arg Ser Leu Pro Ser Ile Ile305 310 315 320Arg Asn Ala Leu Ser Glu Asp Ser Val Gly Arg Asp Ser Lys Thr Phe 325 330 335Thr Pro Ser Thr Thr Asp Thr Ser Thr Arg Lys Ser Gln Ala Val 340 345 3505352PRTRattus norvegicus 5Met Asp Pro Ile Ser Asn Asp Ser Ser Glu Ile Thr Tyr Asp Tyr Ser1 5 10 15Asp Gly Thr Pro Asn Pro Asp Met Pro Ala Asp Gly Val Tyr Ile Pro 20 25 30Lys Met Glu Pro Gly Asp Ile Ala Ala Leu Ile Ile Tyr Leu Ala Val 35 40 45Phe Leu Val Gly Val Thr Gly Asn Ala Leu Val Val Trp Val Thr Ala 50 55 60Phe Glu Ala Lys Arg Thr Val Asn Ala Ile Trp Phe Leu Asn Leu Ala65 70 75 80Val Ala Asp Leu Leu Ser Cys Leu Ala Leu Pro Ile Leu Phe Thr Ser 85 90 95Ile Val Lys His Asn His Trp Pro Phe Gly Asp Gln Ala Cys Ile Val 100 105 110Leu Pro Ser Leu Ile Leu Leu Asn Met Tyr Ser Ser Ile Leu Leu Leu 115 120 125Ala Thr Ile Ser Ala Asp Arg Phe Leu Leu Val Phe Lys Pro Ile Trp 130 135 140Cys Gln Lys Phe Arg Arg Pro Gly Leu Ala Trp Met Ala Cys Gly Val145 150 155 160Thr Trp Val Leu Ala Leu Leu Leu Thr Ile Pro Ser Phe Val Phe Arg 165 170 175Arg Ile His Lys Asp Pro Tyr Ser Asp Ser Ile Leu Cys Asn Ile Asp 180 185 190Tyr Ser Lys Gly Pro Phe Phe Ile Glu Lys Ala Ile Ala Ile Leu Arg 195 200 205Leu Met Val Gly Phe Val Leu Pro Leu Leu Thr Leu Asn Ile Cys Tyr 210 215 220Thr Phe Leu Leu Ile Arg Thr Trp Ser Arg Lys Ala Thr Arg Ser Thr225 230 235 240Lys Thr Leu Lys Val Val Met Ala Val Val Thr Cys Phe Phe Val Phe 245 250 255Trp Leu Pro Tyr Gln Val Thr Gly Val Ile Leu Ala Trp Leu Pro Arg 260 265 270Ser Ser Ser Thr Phe Gln Ser Val Glu Arg Leu Asn Ser Leu Cys Val 275 280 285Ser Leu Ala Tyr Ile Asn Cys Cys Val Asn Pro Ile Ile Tyr Val Met 290 295 300Ala Gly Gln Gly Phe His Gly Arg Leu Arg Arg Ser Leu Pro Ser Ile305 310 315 320Ile Arg Asn Val Leu Ser Glu Asp Ser Leu Gly Arg Asp Ser Lys Ser 325 330 335Phe Thr Arg Ser Thr Met Asp Thr Ser Thr Gln Lys Ser Gln Ala Val 340 345 350674PRTHomo sapiens 6Thr Leu Gln Lys Lys Ile Glu Glu Ile Ala Ala Lys Tyr Lys His Ser1 5 10 15Val Val Lys Lys Cys Cys Tyr Asp Gly Ala Cys Val Asn Asn Asp Glu 20 25 30Thr Cys Glu Gln Arg Ala Ala Arg Ile Ser Leu Gly Pro Arg Cys Ile 35 40 45Lys Ala Phe Thr Glu Cys Cys Val Val Ala Ser Gln Leu Arg Ala Asn 50 55 60Ile Ser His Lys Asp Met Gln Leu Gly Arg65 707337PRTHomo sapiens 7Met Gly Asn Asp Ser Val Ser Tyr Glu Tyr Gly Asp Tyr Ser Asp Leu1 5 10 15Ser Asp Arg Pro Val Asp Cys Leu Asp Gly Ala Cys Leu Ala Ile Asp 20 25 30Pro Leu Arg Val Ala Pro Leu Pro Leu Tyr Ala Ala Ile Phe Leu Val 35 40 45Gly Val Pro Gly Asn Ala Met Val Ala Trp Val Ala Gly Lys Val Ala 50 55 60Arg Arg Arg Val Gly Ala Thr Trp Leu Leu His Leu Ala Val Ala Asp65 70 75 80Leu Leu Cys Cys Leu Ser Leu Pro Ile Leu Ala Val Pro Ile Ala Arg 85 90 95Gly Gly His Trp Pro Tyr Gly Ala Val Gly Cys Arg Ala Leu Pro Ser 100 105 110Ile Ile Leu Leu Thr Met Tyr Ala Ser Val Leu Leu Leu Ala Ala Leu 115 120 125Ser Ala Asp Leu Cys Phe Leu Ala Leu Gly Pro Ala Trp Trp Ser Thr 130 135 140Val Gln Arg Ala Cys Gly Val Gln Val Ala Cys Gly Ala Ala Trp Thr145 150 155 160Leu Ala Leu Leu Leu Thr Val Pro Ser Ala Ile Tyr Arg Arg Leu His 165 170 175Gln Glu His Phe Pro Ala Arg Leu Gln Cys Val Val Asp Tyr Gly Gly 180 185 190Ser Ser Ser Thr Glu Asn Ala Val Thr Ala Ile Arg Phe Leu Phe Gly 195 200 205Phe Leu Gly Pro Leu Val Ala Val Ala Ser Cys His Ser Ala Leu Leu 210 215 220Cys Trp Ala Ala Arg Arg Cys Arg Pro Leu Gly Thr Ala Ile Val Val225 230 235 240Gly Phe Phe Val Cys Trp Ala Pro Tyr His Leu Leu Gly Leu Val Leu 245 250 255Thr Val Ala Ala Pro Asn Ser Ala Leu Leu Ala Arg Ala Leu Arg Ala 260 265 270Glu Pro Leu Ile Val Gly Leu Ala Leu Ala His Ser Cys Leu Asn Pro 275 280 285Met Leu Phe Leu Tyr Phe Gly Arg Ala Gln Leu Arg Arg Ser Leu Pro 290 295 300Ala Ala Cys His Trp Ala Leu Arg Glu Ser Gln Gly Gln Asp Glu Ser305 310 315 320Val Asp Ser Lys Lys Ser Thr Ser His Asp Leu Val Ser Glu Met Glu 325 330 335Val8482PRTHomo sapiens 8Met Ala Ser Phe Ser Ala Glu Thr Asn Ser Thr Asp Leu Leu Ser Gln1 5 10 15Pro Trp Asn Glu Pro Pro Val Ile Leu Ser Met Val Ile Leu Ser Leu 20 25 30Thr Phe Leu Leu Gly Leu Pro Gly Asn Gly Leu Val Leu Trp Val Ala 35 40 45Gly Leu Lys Met Gln Arg Thr Val Asn Thr Ile Trp Phe Leu His Leu 50 55 60Thr Leu Ala Asp Leu Leu Cys Cys Leu Ser Leu Pro Phe Ser Leu Ala65 70 75 80His Leu Ala Leu Gln Gly Gln Trp Pro Tyr Gly Arg Phe Leu Cys Lys 85 90 95Leu Ile Pro Ser Ile Ile Val Leu Asn Met Phe Ala Ser Val Phe Leu 100 105 110Leu Thr Ala Ile Ser Leu Asp Arg Cys Leu Val Val Phe Lys Pro Ile 115 120 125Trp Cys Gln Asn His Arg Asn Val Gly Met Ala Cys Ser Ile Cys Gly 130 135 140Cys Ile Trp Val Val Ala Cys Val Met Cys Ile Pro Val Phe Val Tyr145 150 155 160Arg Glu Ile Phe Thr Thr Asp Asn His Asn Arg Cys Gly Tyr Lys Phe 165 170 175Gly Leu Ser Ser Ser Leu Asp Tyr Pro Asp Phe Tyr Gly Asp Pro Leu 180 185 190Glu Asn Arg Ser Leu Glu Asn Ile Val Gln Pro Pro Gly Glu Met Asn 195 200 205Asp Arg Leu Asp Pro Ser Ser Phe Gln Thr Asn Asp His Pro Trp Thr 210 215 220Val Pro Thr Val Phe Gln Pro Gln Thr Phe Gln Arg Pro Ser Ala Asp225 230 235 240Ser Leu Pro Arg Gly Ser Ala Arg Leu Thr Ser Gln Asn Leu Tyr Ser 245 250 255Asn Val Phe Lys Pro Ala Asp Val Val Ser Pro Lys Ile Pro Ser Gly 260 265 270Phe Pro Ile Glu Asp His Glu Thr Ser Pro Leu Asp Asn Ser Asp Ala 275

280 285Phe Leu Ser Thr His Leu Lys Leu Phe Pro Ser Ala Ser Ser Asn Ser 290 295 300Phe Tyr Glu Ser Glu Leu Pro Gln Gly Phe Gln Asp Tyr Tyr Asn Leu305 310 315 320Gly Gln Phe Thr Asp Asp Asp Gln Val Pro Thr Pro Leu Val Ala Ile 325 330 335Thr Ile Thr Arg Leu Val Val Gly Phe Leu Leu Pro Ser Val Ile Met 340 345 350Ile Ala Cys Tyr Ser Phe Ile Val Phe Arg Met Gln Arg Gly Arg Phe 355 360 365Ala Lys Ser Gln Ser Lys Thr Phe Arg Val Ala Val Val Val Val Ala 370 375 380Val Phe Leu Val Cys Trp Thr Pro Tyr His Ile Phe Gly Val Leu Ser385 390 395 400Leu Leu Thr Asp Pro Glu Thr Pro Leu Gly Lys Thr Leu Met Ser Trp 405 410 415Asp His Val Cys Ile Ala Leu Ala Ser Ala Asn Ser Cys Phe Asn Pro 420 425 430Phe Leu Tyr Ala Leu Leu Gly Lys Asp Phe Arg Lys Lys Ala Arg Gln 435 440 445Ser Ile Gln Gly Ile Leu Glu Ala Ala Phe Ser Glu Glu Leu Thr Arg 450 455 460Ser Thr His Cys Pro Ser Asn Asn Val Ile Ser Glu Arg Asn Ser Thr465 470 475 480Thr Val9350PRTHomo sapiens 9Met Glu Thr Asn Ser Ser Leu Pro Thr Asn Ile Ser Gly Gly Thr Pro1 5 10 15Ala Val Ser Ala Gly Tyr Leu Phe Leu Asp Ile Ile Thr Tyr Leu Val 20 25 30Phe Ala Val Thr Phe Val Leu Gly Val Leu Gly Asn Gly Leu Val Ile 35 40 45Trp Val Ala Gly Phe Arg Met Thr His Thr Val Thr Thr Ile Ser Tyr 50 55 60Leu Asn Leu Ala Val Ala Asp Phe Cys Phe Thr Ser Thr Leu Pro Phe65 70 75 80Phe Met Val Arg Lys Ala Met Gly Gly His Trp Pro Phe Gly Trp Phe 85 90 95Leu Cys Lys Phe Leu Phe Thr Ile Val Asp Ile Asn Leu Phe Gly Ser 100 105 110Val Phe Leu Ile Ala Leu Ile Ala Leu Asp Arg Cys Val Cys Val Leu 115 120 125His Pro Val Trp Thr Gln Asn His Arg Thr Val Ser Leu Ala Lys Lys 130 135 140Val Ile Ile Gly Pro Trp Val Met Ala Leu Leu Leu Thr Leu Pro Val145 150 155 160Ile Ile Arg Val Thr Thr Val Pro Gly Lys Thr Gly Thr Val Ala Cys 165 170 175Thr Phe Asn Phe Ser Pro Trp Thr Asn Asp Pro Lys Glu Arg Ile Asn 180 185 190Val Ala Val Ala Met Leu Thr Val Arg Gly Ile Ile Arg Phe Ile Ile 195 200 205Gly Phe Ser Ala Pro Met Ser Ile Val Ala Val Ser Tyr Gly Leu Ile 210 215 220Ala Thr Lys Ile His Lys Gln Gly Leu Ile Lys Ser Ser Pro Pro Leu225 230 235 240Arg Val Leu Ser Phe Val Ala Ala Ala Phe Phe Leu Cys Trp Ser Pro 245 250 255Tyr Gln Val Val Ala Leu Ile Ala Thr Val Arg Ile Arg Glu Leu Leu 260 265 270Gln Gly Met Tyr Lys Glu Ile Gly Ile Ala Val Asp Val Thr Ser Ala 275 280 285Leu Ala Phe Phe Asn Ser Cys Leu Asn Pro Met Leu Tyr Val Phe Met 290 295 300Gly Gln Asp Phe Arg Glu Arg Leu Ile His Ala Leu Pro Ala Ser Leu305 310 315 320Glu Arg Ala Leu Thr Glu Asp Ser Thr Gln Thr Ser Asp Thr Ala Thr 325 330 335Asn Ser Thr Leu Pro Ser Ala Glu Val Ala Leu Gln Ala Lys 340 345 35010373PRTHomo sapiens 10Met Arg Met Glu Asp Glu Asp Tyr Asn Thr Ser Ile Ser Tyr Gly Asp1 5 10 15Glu Tyr Pro Asp Tyr Leu Asp Ser Ile Val Val Leu Glu Asp Leu Ser 20 25 30Pro Leu Glu Ala Arg Val Thr Arg Ile Phe Leu Val Val Val Tyr Ser 35 40 45Ile Val Cys Phe Leu Gly Ile Leu Gly Asn Gly Leu Val Ile Ile Ile 50 55 60Ala Thr Phe Lys Met Lys Lys Thr Val Asn Met Val Trp Phe Leu Asn65 70 75 80Leu Ala Val Ala Asp Phe Leu Phe Asn Val Phe Leu Pro Ile His Ile 85 90 95Thr Tyr Ala Ala Met Asp Tyr His Trp Val Phe Gly Thr Ala Met Cys 100 105 110Lys Ile Ser Asn Phe Leu Leu Ile His Asn Met Phe Thr Ser Val Phe 115 120 125Leu Leu Thr Ile Ile Ser Ser Asp Arg Cys Ile Ser Val Leu Leu Pro 130 135 140Val Trp Ser Gln Asn His Arg Ser Val Arg Leu Ala Tyr Met Ala Cys145 150 155 160Met Val Ile Trp Val Leu Ala Phe Phe Leu Ser Ser Pro Ser Leu Val 165 170 175Phe Arg Asp Thr Ala Asn Leu His Gly Lys Ile Ser Cys Phe Asn Asn 180 185 190Phe Ser Leu Ser Thr Pro Gly Ser Ser Ser Trp Pro Thr His Ser Gln 195 200 205Met Asp Pro Val Gly Tyr Ser Arg His Met Val Val Thr Val Thr Arg 210 215 220Phe Leu Cys Gly Phe Leu Val Pro Val Leu Ile Ile Thr Ala Cys Tyr225 230 235 240Leu Thr Ile Val Cys Lys Leu His Arg Asn Arg Leu Ala Lys Thr Lys 245 250 255Lys Pro Phe Lys Ile Ile Val Thr Ile Ile Ile Thr Phe Phe Leu Cys 260 265 270Trp Cys Pro Tyr His Thr Leu Asn Leu Leu Glu Leu His His Thr Ala 275 280 285Met Pro Gly Ser Val Phe Ser Leu Gly Leu Pro Leu Ala Thr Ala Leu 290 295 300Ala Ile Ala Asn Ser Cys Met Asn Pro Ile Leu Tyr Val Phe Met Gly305 310 315 320Gln Asp Phe Lys Lys Phe Lys Val Ala Leu Phe Ser Arg Leu Val Asn 325 330 335Ala Leu Ser Glu Asp Thr Gly His Ser Ser Tyr Pro Ser His Arg Ser 340 345 350Phe Thr Lys Met Ser Ser Met Asn Glu Arg Thr Ser Met Asn Glu Arg 355 360 365Glu Thr Gly Met Leu 37011216PRTHomo sapiens 11Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro1 5 10 15Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val 20 25 30Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val 35 40 45Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln 50 55 60Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln65 70 75 80Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala 85 90 95Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro 100 105 110Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr 115 120 125Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser 130 135 140Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr145 150 155 160Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr 165 170 175Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe 180 185 190Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys 195 200 205Ser Leu Ser Leu Ser Pro Gly Lys 210 2151231PRTHomo sapiens 12Met Asp Ser Phe Asn Tyr Thr Thr Pro Asp Tyr Gly His Tyr Asp Asp1 5 10 15Lys Asp Thr Leu Asp Leu Asn Thr Pro Val Asp Lys Thr Ser Asn 20 25 301337PRTHomo sapiens 13Met Asp Ser Phe Asn Tyr Thr Thr Pro Asp Tyr Gly His Tyr Asp Asp1 5 10 15Lys Asp Thr Leu Asp Leu Asn Thr Pro Val Asp Lys Thr Ser Asn Thr 20 25 30Leu Arg Val Pro Asp 351437PRTMacaca fascicularis 14Met Asp Pro Phe Ser Ser Thr Thr Leu Asp Tyr Glu His Tyr Asp Gly1 5 10 15Lys Asn Val Leu Asp Ser Asp Thr Pro Val Asp Lys Thr Ser Asn Thr 20 25 30Leu Arg Val Pro Asp 3515398PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 15Ala Glu Ser His Leu Ser Leu Leu Tyr His Leu Thr Ala Val Ser Ser1 5 10 15Pro Ala Pro Gly Thr Pro Ala Phe Trp Val Ser Gly Trp Leu Gly Pro 20 25 30Gln Gln Tyr Leu Ser Tyr Asn Ser Leu Arg Gly Glu Ala Glu Pro Cys 35 40 45Gly Ala Trp Val Trp Glu Asn Gln Val Ser Trp Tyr Trp Glu Lys Glu 50 55 60Thr Thr Asp Leu Arg Ile Lys Glu Lys Leu Phe Leu Glu Ala Phe Lys65 70 75 80Ala Leu Gly Gly Lys Gly Pro Tyr Thr Leu Gln Gly Leu Leu Gly Cys 85 90 95Glu Leu Gly Pro Asp Asn Thr Ser Val Pro Thr Ala Lys Phe Ala Leu 100 105 110Asn Gly Glu Glu Phe Met Asn Phe Asp Leu Lys Gln Gly Thr Trp Gly 115 120 125Gly Asp Trp Pro Glu Ala Leu Ala Ile Ser Gln Arg Trp Gln Gln Gln 130 135 140Asp Lys Ala Ala Asn Lys Glu Leu Thr Phe Leu Leu Phe Ser Cys Pro145 150 155 160His Arg Leu Arg Glu His Leu Glu Arg Gly Arg Gly Asn Leu Glu Trp 165 170 175Lys Glu Pro Pro Ser Met Arg Leu Lys Ala Arg Pro Ser Ser Pro Gly 180 185 190Phe Ser Val Leu Thr Cys Ser Ala Phe Ser Phe Tyr Pro Pro Glu Leu 195 200 205Gln Leu Arg Phe Leu Arg Asn Gly Leu Ala Ala Gly Thr Gly Gln Gly 210 215 220Asp Phe Gly Pro Asn Ser Asp Gly Ser Phe His Ala Ser Ser Ser Leu225 230 235 240Thr Val Lys Ser Gly Asp Glu His His Tyr Cys Cys Ile Val Gln His 245 250 255Ala Gly Leu Ala Gln Pro Leu Arg Val Glu Leu Glu Ser Pro Ala Lys 260 265 270Ser Ser Val Asn Ser Arg Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys 275 280 285Ile Glu Trp His Glu His His His His His His Ile Gln Arg Thr Pro 290 295 300Lys Ile Gln Val Tyr Ser Arg His Pro Ala Glu Asn Gly Lys Ser Asn305 310 315 320Phe Leu Asn Cys Tyr Val Ser Gly Phe His Pro Ser Asp Ile Glu Val 325 330 335Asp Leu Leu Lys Asn Gly Glu Arg Ile Glu Lys Val Glu His Ser Asp 340 345 350Leu Ser Phe Ser Lys Asp Trp Ser Phe Tyr Leu Leu Tyr Tyr Thr Glu 355 360 365Phe Thr Pro Thr Glu Lys Asp Glu Tyr Ala Cys Arg Val Asn His Val 370 375 380Thr Leu Ser Gln Pro Lys Ile Val Lys Trp Asp Arg Asp Met385 390 39516398PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 16Ala Glu Ser His Leu Ser Leu Leu Tyr His Leu Thr Ala Val Ser Ser1 5 10 15Pro Ala Pro Gly Thr Pro Ala Phe Trp Val Ser Gly Trp Leu Gly Pro 20 25 30Gln Gln Tyr Leu Ser Tyr Asp Ser Leu Arg Gly Gln Ala Glu Pro Cys 35 40 45Gly Ala Trp Val Trp Glu Asn Gln Val Ser Trp Tyr Trp Glu Lys Glu 50 55 60Thr Thr Asp Leu Arg Ile Lys Glu Lys Leu Phe Leu Glu Ala Phe Lys65 70 75 80Ala Leu Gly Gly Lys Gly Pro Tyr Thr Leu Gln Gly Leu Leu Gly Cys 85 90 95Glu Leu Ser Pro Asp Asn Thr Ser Val Pro Thr Ala Lys Phe Ala Leu 100 105 110Asn Gly Glu Glu Phe Met Asn Phe Asp Leu Lys Gln Gly Thr Trp Gly 115 120 125Gly Asp Trp Pro Glu Ala Leu Ala Ile Ser Gln Arg Trp Gln Gln Gln 130 135 140Asp Lys Ala Ala Asn Lys Glu Leu Thr Phe Leu Leu Phe Ser Cys Pro145 150 155 160His Arg Leu Arg Glu His Leu Glu Arg Gly Arg Gly Asn Leu Glu Trp 165 170 175Lys Glu Pro Pro Ser Met Arg Leu Lys Ala Arg Pro Gly Asn Pro Gly 180 185 190Phe Ser Val Leu Thr Cys Ser Ala Phe Ser Phe Tyr Pro Pro Glu Leu 195 200 205Gln Leu Arg Phe Leu Arg Asn Gly Met Ala Ala Gly Thr Gly Gln Gly 210 215 220Asp Phe Gly Pro Asn Ser Asp Gly Ser Phe His Ala Ser Ser Ser Leu225 230 235 240Thr Val Lys Ser Gly Asp Glu His His Tyr Cys Cys Ile Val Gln His 245 250 255Ala Gly Leu Ala Gln Pro Leu Arg Val Glu Leu Glu Thr Pro Ala Lys 260 265 270Ser Ser Val Asn Ser Arg Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys 275 280 285Ile Glu Trp His Glu His His His His His His Ile Gln Arg Thr Pro 290 295 300Lys Ile Gln Val Tyr Ser Arg His Pro Pro Glu Asn Gly Lys Pro Asn305 310 315 320Phe Leu Asn Cys Tyr Val Ser Gly Phe His Pro Ser Asp Ile Glu Val 325 330 335Asp Leu Leu Lys Asn Gly Glu Lys Met Gly Lys Val Glu His Ser Asp 340 345 350Leu Ser Phe Ser Lys Asp Trp Ser Phe Tyr Leu Leu Tyr Tyr Thr Glu 355 360 365Phe Thr Pro Asn Glu Lys Asp Glu Tyr Ala Cys Arg Val Asn His Val 370 375 380Thr Leu Ser Gly Pro Arg Thr Val Lys Trp Asp Arg Asp Met385 390 39517400PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 17Ser Glu Thr Arg Pro Pro Leu Met Tyr His Leu Thr Ala Val Ser Asn1 5 10 15Pro Ser Thr Gly Leu Pro Ser Phe Trp Ala Thr Gly Trp Leu Gly Pro 20 25 30Gln Gln Tyr Leu Thr Tyr Asn Ser Leu Arg Gln Glu Ala Asp Pro Cys 35 40 45Gly Ala Trp Met Trp Glu Asn Gln Val Ser Trp Tyr Trp Glu Lys Glu 50 55 60Thr Thr Asp Leu Lys Ser Lys Glu Gln Leu Phe Leu Glu Ala Leu Lys65 70 75 80Thr Leu Glu Lys Ile Leu Asn Gly Thr Tyr Thr Leu Gln Gly Leu Leu 85 90 95Gly Cys Glu Leu Ala Ser Asp Asn Ser Ser Val Pro Thr Ala Val Phe 100 105 110Ala Leu Asn Gly Glu Glu Phe Met Lys Phe Asn Pro Arg Ile Gly Asn 115 120 125Trp Thr Gly Glu Trp Pro Glu Thr Glu Ile Val Ala Asn Leu Trp Met 130 135 140Lys Gln Pro Asp Ala Ala Arg Lys Glu Ser Glu Phe Leu Leu Asn Ser145 150 155 160Cys Pro Glu Arg Leu Leu Gly His Leu Glu Arg Gly Arg Arg Asn Leu 165 170 175Glu Trp Lys Glu Pro Pro Ser Met Arg Leu Lys Ala Arg Pro Gly Asn 180 185 190Ser Gly Ser Ser Val Leu Thr Cys Ala Ala Phe Ser Phe Tyr Pro Pro 195 200 205Glu Leu Lys Phe Arg Phe Leu Arg Asn Gly Leu Ala Ser Gly Ser Gly 210 215 220Asn Cys Ser Thr Gly Pro Asn Gly Asp Gly Ser Phe His Ala Trp Ser225 230 235 240Leu Leu Glu Val Lys Arg Gly Asp Glu His His Tyr Gln Cys Gln Val 245 250 255Glu His Glu Gly Leu Ala Gln Pro Leu Thr Val Asp Leu Asp Ser Ser 260 265 270Ala Arg Ser Ser Val Asn Ser Arg Gly Leu Asn Asp Ile Phe Glu Ala 275 280 285Gln Lys Ile Glu Trp His Glu His His His His His His Ile Gln Lys 290 295 300Thr Pro Gln Ile Gln Val Tyr Ser Arg His Pro Pro Glu Asn Gly Lys305 310 315 320Pro Asn Ile Leu Asn Cys Tyr Val Thr Gln Phe His Pro Pro His Ile 325 330 335Glu Ile Gln Met Leu Lys Asn Gly Lys Lys Ile Pro Lys Val Glu Met 340 345 350Ser Asp Met Ser Phe Ser Lys Asp Trp Ser Phe Tyr Ile Leu Ala His 355 360 365Thr Glu Phe Thr Pro Thr Glu Thr Asp

Thr Tyr Ala Cys Arg Val Lys 370 375 380His Asp Ser Met Ala Glu Pro Lys Thr Val Tyr Trp Asp Arg Asp Met385 390 395 40018274PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 18Gln Val Asp Thr Thr Lys Ala Val Ile Thr Leu Gln Pro Pro Trp Val1 5 10 15Ser Val Phe Gln Glu Glu Thr Val Thr Leu His Cys Glu Val Leu His 20 25 30Leu Pro Gly Ser Ser Ser Thr Gln Trp Phe Leu Asn Gly Thr Ala Thr 35 40 45Gln Thr Ser Thr Pro Ser Tyr Arg Ile Thr Ser Ala Ser Val Asn Asp 50 55 60Ser Gly Glu Tyr Arg Cys Gln Arg Gly Leu Ser Gly Arg Ser Asp Pro65 70 75 80Ile Gln Leu Glu Ile His Arg Gly Trp Leu Leu Leu Gln Val Ser Ser 85 90 95Arg Val Phe Thr Glu Gly Glu Pro Leu Ala Leu Arg Cys His Ala Trp 100 105 110Lys Asp Lys Leu Val Tyr Asn Val Leu Tyr Tyr Arg Asn Gly Lys Ala 115 120 125Phe Lys Phe Phe His Trp Asn Ser Asn Leu Thr Ile Leu Lys Thr Asn 130 135 140Ile Ser His Asn Gly Thr Tyr His Cys Ser Gly Met Gly Lys His Arg145 150 155 160Tyr Thr Ser Ala Gly Ile Ser Val Thr Val Lys Glu Leu Phe Pro Ala 165 170 175Pro Val Leu Asn Ala Ser Val Thr Ser Pro Leu Leu Glu Gly Asn Leu 180 185 190Val Thr Leu Ser Cys Glu Thr Lys Leu Leu Leu Gln Arg Pro Gly Leu 195 200 205Gln Leu Tyr Phe Ser Phe Tyr Met Gly Ser Lys Thr Leu Arg Gly Arg 210 215 220Asn Thr Ser Ser Glu Tyr Gln Ile Leu Thr Ala Arg Arg Glu Asp Ser225 230 235 240Gly Leu Tyr Trp Cys Glu Ala Ala Thr Glu Asp Gly Asn Val Leu Lys 245 250 255Arg Ser Pro Glu Leu Glu Leu Gln Val Asn Ser Arg His His His His 260 265 270His His19186PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 19Gln Ala Ala Ala Pro Pro Lys Ala Val Leu Lys Leu Glu Pro Pro Trp1 5 10 15Ile Asn Val Leu Gln Glu Asp Ser Val Thr Leu Thr Cys Gln Gly Ala 20 25 30Arg Ser Pro Glu Ser Asp Ser Ile Gln Trp Phe His Asn Gly Asn Leu 35 40 45Ile Pro Thr His Thr Gln Pro Ser Tyr Arg Phe Lys Ala Asn Asn Asn 50 55 60Asp Ser Gly Glu Tyr Thr Cys Gln Thr Gly Gln Thr Ser Leu Ser Asp65 70 75 80Pro Val His Leu Thr Val Leu Ser Glu Trp Leu Val Leu Gln Thr Pro 85 90 95His Leu Glu Phe Gln Glu Gly Glu Thr Ile Met Leu Arg Cys His Ser 100 105 110Trp Lys Asp Lys Pro Leu Val Lys Val Thr Phe Phe Gln Asn Gly Lys 115 120 125Ser Gln Lys Phe Ser His Leu Asp Pro Thr Phe Ser Ile Pro Gln Ala 130 135 140Asn His Ser His Ser Gly Asp Tyr His Cys Thr Gly Asn Ile Gly Tyr145 150 155 160Thr Leu Phe Ser Ser Lys Pro Val Thr Ile Thr Val Gln Val Pro Ser 165 170 175Val Asn Ser Arg His His His His His His 180 18520186PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 20Gln Ala Ala Ala Pro Pro Lys Ala Val Leu Lys Leu Glu Pro Pro Trp1 5 10 15Ile Asn Val Leu Gln Glu Asp Ser Val Thr Leu Thr Cys Gln Gly Ala 20 25 30Arg Ser Pro Glu Ser Asp Ser Ile Gln Trp Phe His Asn Gly Asn Leu 35 40 45Ile Pro Thr His Thr Gln Pro Ser Tyr Arg Phe Lys Ala Asn Asn Asn 50 55 60Asp Ser Gly Glu Tyr Thr Cys Gln Thr Gly Gln Thr Ser Leu Ser Asp65 70 75 80Pro Val His Leu Thr Val Leu Ser Glu Trp Leu Val Leu Gln Thr Pro 85 90 95His Leu Glu Phe Gln Glu Gly Glu Thr Ile Met Leu Arg Cys His Ser 100 105 110Trp Lys Asp Lys Pro Leu Val Lys Val Thr Phe Phe Gln Asn Gly Lys 115 120 125Ser Gln Lys Phe Ser Arg Leu Asp Pro Thr Phe Ser Ile Pro Gln Ala 130 135 140Asn His Ser His Ser Gly Asp Tyr His Cys Thr Gly Asn Ile Gly Tyr145 150 155 160Thr Leu Phe Ser Ser Lys Pro Val Thr Ile Thr Val Gln Val Pro Ser 165 170 175Val Asn Ser Arg His His His His His His 180 18521187PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 21Gly Met Arg Thr Glu Asp Leu Pro Lys Ala Val Val Phe Leu Glu Pro1 5 10 15Gln Trp Tyr Arg Val Leu Glu Lys Asp Ser Val Thr Leu Lys Cys Gln 20 25 30Gly Ala Tyr Ser Pro Glu Asp Asn Ser Thr Gln Trp Phe His Asn Glu 35 40 45Ser Leu Ile Ser Ser Gln Ala Ser Ser Tyr Phe Ile Asp Ala Ala Thr 50 55 60Val Asp Asp Ser Gly Glu Tyr Arg Cys Gln Thr Asn Leu Ser Thr Leu65 70 75 80Ser Asp Pro Val Gln Leu Glu Val His Ile Gly Trp Leu Leu Leu Gln 85 90 95Ala Pro Arg Trp Val Phe Lys Glu Glu Asp Pro Ile His Leu Arg Cys 100 105 110His Ser Trp Lys Asn Thr Ala Leu His Lys Val Thr Tyr Leu Gln Asn 115 120 125Gly Lys Gly Arg Lys Tyr Phe His His Asn Ser Asp Phe Tyr Ile Pro 130 135 140Lys Ala Thr Leu Lys Asp Ser Gly Ser Tyr Phe Cys Arg Gly Leu Phe145 150 155 160Gly Ser Lys Asn Val Ser Ser Glu Thr Val Asn Ile Thr Ile Thr Gln 165 170 175Gly Val Asn Ser Arg His His His His His His 180 18522187PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 22Gly Met Arg Thr Glu Asp Leu Pro Lys Ala Val Val Phe Leu Glu Pro1 5 10 15Gln Trp Tyr Arg Val Leu Glu Lys Asp Ser Val Thr Leu Lys Cys Gln 20 25 30Gly Ala Tyr Ser Pro Glu Asp Asn Ser Thr Gln Trp Phe His Asn Glu 35 40 45Ser Leu Ile Ser Ser Gln Ala Ser Ser Tyr Phe Ile Asp Ala Ala Thr 50 55 60Val Asp Asp Ser Gly Glu Tyr Arg Cys Gln Thr Asn Leu Ser Thr Leu65 70 75 80Ser Asp Pro Val Gln Leu Glu Val His Ile Gly Trp Leu Leu Leu Gln 85 90 95Ala Pro Arg Trp Val Phe Lys Glu Glu Asp Pro Ile His Leu Arg Cys 100 105 110His Ser Trp Lys Asn Thr Ala Leu His Lys Val Thr Tyr Leu Gln Asn 115 120 125Gly Lys Gly Arg Lys Tyr Phe His His Asn Ser Asp Phe Tyr Ile Pro 130 135 140Lys Ala Thr Leu Lys Asp Ser Gly Ser Tyr Phe Cys Arg Gly Leu Val145 150 155 160Gly Ser Lys Asn Val Ser Ser Glu Thr Val Asn Ile Thr Ile Thr Gln 165 170 175Gly Val Asn Ser Arg His His His His His His 180 18523186PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 23Thr Pro Ala Ala Pro Pro Lys Ala Val Leu Lys Leu Glu Pro Gln Trp1 5 10 15Ile Asn Val Leu Gln Glu Asp Ser Val Thr Leu Thr Cys Arg Gly Thr 20 25 30His Ser Pro Glu Ser Asp Ser Ile Gln Trp Phe His Asn Gly Asn Leu 35 40 45Ile Pro Thr His Thr Gln Pro Ser Tyr Arg Phe Lys Ala Asn Asn Asn 50 55 60Asp Ser Gly Glu Tyr Thr Cys Gln Thr Gly Gln Thr Ser Leu Ser Asp65 70 75 80Pro Val His Leu Thr Val Leu Ser Glu Trp Leu Val Leu Gln Thr Pro 85 90 95His Leu Glu Phe Gln Glu Gly Glu Thr Ile Val Leu Arg Cys His Ser 100 105 110Trp Lys Asp Lys Pro Leu Val Lys Val Thr Phe Phe Gln Asn Gly Lys 115 120 125Ser Lys Lys Phe Ser Arg Ser Asp Pro Asn Phe Ser Ile Pro Gln Ala 130 135 140Asn His Ser His Ser Gly Asp Tyr His Cys Thr Gly Asn Ile Gly Tyr145 150 155 160Thr Leu Tyr Ser Ser Lys Pro Val Thr Ile Thr Val Gln Ala Pro Ser 165 170 175Asp Asn Ser Arg His His His His His His 180 18524907PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 24Asp Gly Ser His His His His His His Gly Thr Met Asp Ser Phe Asn1 5 10 15Tyr Thr Thr Pro Asp Tyr Gly His Tyr Asp Asp Lys Asp Thr Leu Asp 20 25 30Leu Asn Thr Pro Val Asp Lys Thr Ser Asn Thr Leu Arg Val Pro Asp 35 40 45Ile Leu Ala Leu Val Ile Phe Ala Val Val Phe Leu Val Gly Val Leu 50 55 60Gly Asn Ala Leu Val Val Trp Val Thr Ala Phe Glu Ala Lys Arg Thr65 70 75 80Ile Asn Ala Ile Trp Phe Leu Asn Leu Ala Val Ala Asp Phe Leu Ser 85 90 95Cys Leu Ala Leu Pro Ile Leu Phe Thr Ser Ile Val Gln His His His 100 105 110Trp Pro Phe Gly Gly Ala Ala Cys Ser Ile Leu Pro Ser Leu Ile Leu 115 120 125Leu Asn Met Tyr Ala Ser Ile Leu Leu Leu Ala Thr Ile Ser Ala Asp 130 135 140Arg Phe Leu Leu Val Phe Lys Pro Ile Trp Cys Gln Asn Phe Arg Gly145 150 155 160Ala Gly Leu Ala Trp Ile Ala Cys Ala Val Ala Trp Gly Leu Ala Leu 165 170 175Leu Leu Thr Ile Pro Ser Phe Leu Tyr Arg Val Val Arg Glu Glu Tyr 180 185 190Phe Pro Pro Lys Val Leu Cys Gly Val Asp Tyr Ser His Asp Lys Arg 195 200 205Arg Glu Arg Ala Val Ala Ile Val Arg Leu Val Leu Gly Phe Leu Trp 210 215 220Pro Leu Leu Thr Leu Thr Ile Cys Tyr Thr Phe Ile Leu Leu Arg Thr225 230 235 240Trp Ser Arg Arg Ala Thr Arg Ser Thr Lys Thr Leu Lys Val Val Val 245 250 255Ala Val Val Ala Ser Phe Phe Ile Phe Trp Leu Pro Tyr Gln Val Thr 260 265 270Gly Ile Met Met Ser Phe Leu Glu Pro Ser Ser Pro Thr Phe Leu Leu 275 280 285Leu Lys Lys Leu Asp Ser Leu Cys Val Ser Phe Ala Tyr Ile Asn Cys 290 295 300Cys Ile Asn Pro Ile Ile Tyr Val Val Ala Gly Gln Gly Phe Gln Gly305 310 315 320Arg Leu Arg Lys Ser Leu Pro Ser Leu Leu Arg Asn Val Leu Thr Glu 325 330 335Glu Ser Val Val Arg Glu Ser Lys Ser Phe Thr Arg Ser Thr Val Asp 340 345 350Thr Met Ala Gln Lys Thr Gln Ala Val Asp Ile Asp Tyr Lys Asp Asp 355 360 365Asp Asp Lys Ile Glu Gly Arg Met Asp Gly Ala Arg Ala Ser Val Leu 370 375 380Ser Gly Gly Glu Leu Asp Arg Trp Glu Lys Ile Arg Leu Arg Pro Gly385 390 395 400Gly Lys Lys Lys Tyr Lys Leu Lys His Ile Val Trp Ala Ser Arg Glu 405 410 415Leu Glu Arg Phe Ala Val Asn Pro Gly Leu Leu Glu Thr Ser Glu Gly 420 425 430Cys Arg Gln Ile Leu Gly Gln Leu Gln Pro Ser Leu Gln Thr Gly Ser 435 440 445Glu Glu Leu Arg Ser Leu Tyr Asn Thr Val Ala Thr Leu Tyr Cys Val 450 455 460His Gln Arg Ile Glu Ile Lys Asp Thr Lys Glu Ala Leu Asp Lys Ile465 470 475 480Glu Glu Glu Gln Asn Lys Ser Lys Lys Lys Ala Gln Gln Ala Ala Ala 485 490 495Asp Thr Gly His Ser Ser Gln Val Ser Gln Asn Tyr Pro Ile Val Gln 500 505 510Asn Ile Gln Gly Gln Met Val His Gln Ala Ile Ser Pro Arg Thr Leu 515 520 525Asn Ala Trp Val Lys Val Val Glu Glu Lys Ala Phe Ser Pro Glu Val 530 535 540Ile Pro Met Phe Ser Ala Leu Ser Glu Gly Ala Thr Pro Gln Asp Leu545 550 555 560Asn Thr Met Leu Asn Thr Val Gly Gly His Gln Ala Ala Met Gln Met 565 570 575Leu Lys Glu Thr Ile Asn Glu Glu Ala Ala Glu Trp Asp Arg Val His 580 585 590Pro Val His Ala Gly Pro Ile Ala Pro Gly Gln Met Arg Glu Pro Arg 595 600 605Gly Ser Asp Ile Ala Gly Thr Thr Ser Thr Leu Gln Glu Gln Ile Gly 610 615 620Trp Met Thr Asn Asn Pro Pro Ile Pro Val Gly Glu Ile Tyr Lys Arg625 630 635 640Trp Ile Ile Leu Gly Leu Asn Lys Ile Val Arg Met Tyr Ser Pro Thr 645 650 655Ser Ile Leu Asp Ile Arg Gln Gly Pro Lys Glu Pro Phe Arg Asp Tyr 660 665 670Val Asp Arg Phe Tyr Lys Thr Leu Arg Ala Glu Gln Ala Ser Gln Glu 675 680 685Val Lys Asn Trp Met Thr Glu Thr Leu Leu Val Gln Asn Ala Asn Pro 690 695 700Asp Cys Lys Thr Ile Leu Lys Ala Leu Gly Pro Ala Ala Thr Leu Glu705 710 715 720Glu Met Met Thr Ala Cys Gln Gly Val Gly Gly Pro Gly His Lys Ala 725 730 735Arg Val Leu Ala Glu Ala Met Ser Gln Val Thr Asn Thr Ala Thr Ile 740 745 750Met Met Gln Arg Gly Asn Phe Arg Asn Gln Arg Lys Met Val Lys Cys 755 760 765Phe Asn Cys Gly Lys Glu Gly His Thr Ala Arg Asn Cys Arg Ala Pro 770 775 780Arg Lys Lys Gly Cys Trp Lys Cys Gly Lys Glu Gly His Gln Met Lys785 790 795 800Asp Cys Thr Glu Arg Gln Ala Asn Phe Leu Gly Lys Ile Trp Pro Ser 805 810 815Tyr Lys Gly Arg Pro Gly Asn Phe Leu Gln Ser Arg Pro Glu Pro Thr 820 825 830Ala Pro Pro Phe Leu Gln Ser Arg Pro Glu Pro Thr Ala Pro Pro Glu 835 840 845Glu Ser Phe Arg Ser Gly Val Glu Thr Thr Thr Pro Pro Gln Lys Gln 850 855 860Glu Pro Ile Asp Lys Glu Leu Tyr Pro Leu Thr Ser Leu Arg Ser Leu865 870 875 880Phe Gly Asn Asp Pro Ser Ser Gln Val Asn Ser Arg Gly Leu Asn Asp 885 890 895Ile Phe Glu Ala Gln Lys Ile Glu Trp His Glu 900 90525874PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 25Asp Gly Ser His His His His His His Gly Thr Met Asn Ser Phe Asn1 5 10 15Tyr Thr Thr Pro Asp Tyr Gly His Tyr Asp Asp Lys Asp Thr Leu Asp 20 25 30Leu Asn Thr Pro Val Asp Lys Thr Ser Asn Thr Leu Arg Val Pro Asp 35 40 45Ile Leu Ala Leu Val Ile Phe Ala Val Val Phe Leu Val Gly Val Leu 50 55 60Gly Asn Ala Leu Val Val Trp Val Thr Ala Phe Glu Ala Lys Arg Thr65 70 75 80Ile Asn Ala Ile Trp Phe Leu Asn Leu Ala Val Ala Asp Phe Leu Ser 85 90 95Cys Leu Ala Leu Pro Ile Leu Phe Thr Ser Ile Val Gln His His His 100 105 110Trp Pro Phe Gly Gly Ala Ala Cys Ser Ile Leu Pro Ser Leu Ile Leu 115 120 125Leu Asn Met Tyr Ala Ser Ile Leu Leu Leu Ala Thr Ile Ser Ala Asp 130 135 140Arg Phe Leu Leu Val Phe Lys Pro Ile Trp Cys Gln Asn Phe Arg Gly145 150 155 160Ala Gly Leu Ala Trp Ile Ala Cys Ala Val Ala Trp Gly Leu Ala Leu 165 170 175Leu Leu Thr Ile Pro Ser Phe Leu Tyr Arg Val Val Arg Glu Glu Tyr 180 185 190Phe Pro Pro Lys Val Leu Cys Gly Val Asp Tyr Ser His Asp Lys Arg 195 200 205Arg Glu Arg Ala

Val Ala Ile Val Arg Leu Val Leu Gly Phe Leu Trp 210 215 220Pro Leu Leu Thr Leu Thr Ile Cys Tyr Thr Phe Ile Leu Leu Arg Thr225 230 235 240Trp Ser Arg Arg Ala Thr Arg Ser Thr Lys Thr Leu Lys Val Val Val 245 250 255Ala Val Val Ala Ser Phe Phe Ile Phe Trp Leu Pro Tyr Gln Val Thr 260 265 270Gly Ile Met Met Ser Phe Leu Glu Pro Ser Ser Pro Thr Phe Leu Leu 275 280 285Leu Asn Lys Leu Asp Ser Leu Cys Val Ser Phe Ala Tyr Ile Asn Cys 290 295 300Cys Ile Asn Pro Ile Ile Tyr Val Val Ala Gly Gln Gly Phe Gln Gly305 310 315 320Arg Leu Arg Lys Ser Leu Pro Ser Leu Leu Arg Asn Val Leu Thr Glu 325 330 335Glu Ser Val Val Arg Glu Ser Lys Ser Phe Thr Arg Ser Thr Val Asp 340 345 350Thr Met Ala Gln Lys Thr Gln Ala Val Asp Ile Gly Ala Arg Ala Ser 355 360 365Val Leu Ser Gly Gly Glu Leu Asp Arg Trp Glu Lys Ile Arg Leu Arg 370 375 380Pro Gly Gly Lys Lys Lys Tyr Lys Leu Lys His Ile Val Trp Ala Ser385 390 395 400Arg Glu Leu Glu Arg Phe Ala Val Asn Pro Gly Leu Leu Glu Thr Ser 405 410 415Glu Gly Cys Arg Gln Ile Leu Gly Gln Leu Gln Pro Ser Leu Gln Thr 420 425 430Gly Ser Glu Glu Leu Arg Ser Leu Tyr Asn Thr Val Ala Thr Leu Tyr 435 440 445Cys Val His Gln Arg Ile Glu Ile Lys Asp Thr Lys Glu Ala Leu Asp 450 455 460Lys Ile Glu Glu Glu Gln Asn Lys Ser Lys Lys Lys Ala Gln Gln Ala465 470 475 480Ala Ala Asp Thr Gly His Ser Ser Gln Val Ser Gln Asn Tyr Pro Ile 485 490 495Val Gln Asn Ile Gln Gly Gln Met Val His Gln Ala Ile Ser Pro Arg 500 505 510Thr Leu Asn Ala Trp Val Lys Val Val Glu Glu Lys Ala Phe Ser Pro 515 520 525Glu Val Ile Pro Met Phe Ser Ala Leu Ser Glu Gly Ala Thr Pro Gln 530 535 540Asp Leu Asn Thr Met Leu Asn Thr Val Gly Gly His Gln Ala Ala Met545 550 555 560Gln Met Leu Lys Glu Thr Ile Asn Glu Glu Ala Ala Glu Trp Asp Arg 565 570 575Val His Pro Val His Ala Gly Pro Ile Ala Pro Gly Gln Met Arg Glu 580 585 590Pro Arg Gly Ser Asp Ile Ala Gly Thr Thr Ser Thr Leu Gln Glu Gln 595 600 605Ile Gly Trp Met Thr Asn Asn Pro Pro Ile Pro Val Gly Glu Ile Tyr 610 615 620Lys Arg Trp Ile Ile Leu Gly Leu Asn Lys Ile Val Arg Met Tyr Ser625 630 635 640Pro Thr Ser Ile Leu Asp Ile Arg Gln Gly Pro Lys Glu Pro Phe Arg 645 650 655Asp Tyr Val Asp Arg Phe Tyr Lys Thr Leu Arg Ala Glu Gln Ala Ser 660 665 670Gln Glu Val Lys Asn Trp Met Thr Glu Thr Leu Leu Val Gln Asn Ala 675 680 685Asn Pro Asp Cys Lys Thr Ile Leu Lys Ala Leu Gly Pro Ala Ala Thr 690 695 700Leu Glu Glu Met Met Thr Ala Cys Gln Gly Val Gly Gly Pro Gly His705 710 715 720Lys Ala Arg Val Leu Ala Glu Ala Met Ser Gln Val Thr Asn Thr Ala 725 730 735Thr Ile Met Met Gln Arg Gly Asn Phe Arg Asn Gln Arg Lys Met Val 740 745 750Lys Cys Phe Asn Cys Gly Lys Glu Gly His Thr Ala Arg Asn Cys Arg 755 760 765Ala Pro Arg Lys Lys Gly Cys Trp Lys Cys Gly Lys Glu Gly His Gln 770 775 780Met Lys Asp Cys Thr Glu Arg Gln Ala Asn Phe Leu Gly Lys Ile Trp785 790 795 800Pro Ser Tyr Lys Gly Arg Pro Gly Asn Phe Leu Gln Ser Arg Pro Glu 805 810 815Pro Thr Ala Pro Pro Phe Leu Gln Ser Arg Pro Glu Pro Thr Ala Pro 820 825 830Pro Glu Glu Ser Phe Arg Ser Gly Val Glu Thr Thr Thr Pro Pro Gln 835 840 845Lys Gln Glu Pro Ile Asp Lys Glu Leu Tyr Pro Leu Thr Ser Leu Arg 850 855 860Ser Leu Phe Gly Asn Asp Pro Ser Ser Gln865 87026908PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 26Asp Gly Ser His His His His His His Gly Thr Met Asp Pro Ile Asp1 5 10 15Asn Ser Ser Phe Glu Ile Asn Tyr Asp His Tyr Gly Thr Met Ala Pro 20 25 30Asn Ile Pro Ala Asp Gly Ile His Leu Pro Lys Arg Gln Pro Gly Asp 35 40 45Val Ala Ala Leu Ile Ile Tyr Ser Val Val Phe Leu Val Gly Val Pro 50 55 60Gly Asn Ala Leu Val Val Trp Val Thr Ala Phe Glu Ala Arg Arg Ala65 70 75 80Val Asn Ala Ile Trp Phe Leu Asn Leu Ala Val Ala Asp Leu Leu Ser 85 90 95Cys Leu Ala Leu Pro Val Leu Phe Thr Thr Val Leu Asn His Asn Tyr 100 105 110Trp Tyr Phe Asp Ala Thr Ala Cys Ile Val Leu Pro Ser Leu Ile Leu 115 120 125Leu Asn Met Tyr Ala Ser Ile Leu Leu Leu Ala Thr Ile Ser Ala Asp 130 135 140Arg Phe Leu Leu Val Phe Lys Pro Ile Trp Cys Gln Lys Val Arg Gly145 150 155 160Thr Gly Leu Ala Trp Met Ala Cys Gly Val Ala Trp Val Leu Ala Leu 165 170 175Leu Leu Thr Ile Pro Ser Phe Val Tyr Arg Glu Ala Tyr Lys Asp Phe 180 185 190Tyr Ser Glu His Thr Val Cys Gly Ile Asn Tyr Gly Gly Gly Ser Phe 195 200 205Pro Lys Glu Lys Ala Val Ala Ile Leu Arg Leu Met Val Gly Phe Val 210 215 220Leu Pro Leu Leu Thr Leu Asn Ile Cys Tyr Thr Phe Leu Leu Leu Arg225 230 235 240Thr Trp Ser Arg Lys Ala Thr Arg Ser Thr Lys Thr Leu Lys Val Val 245 250 255Met Ala Val Val Ile Cys Phe Phe Ile Phe Trp Leu Pro Tyr Gln Val 260 265 270Thr Gly Val Met Ile Ala Trp Leu Pro Pro Ser Ser Pro Thr Leu Lys 275 280 285Arg Val Glu Lys Leu Asn Ser Leu Cys Val Ser Leu Ala Tyr Ile Asn 290 295 300Cys Cys Val Asn Pro Ile Ile Tyr Val Met Ala Gly Gln Gly Phe His305 310 315 320Gly Arg Leu Leu Arg Ser Leu Pro Ser Ile Ile Arg Asn Ala Leu Ser 325 330 335Glu Asp Ser Val Gly Arg Asp Ser Lys Thr Phe Thr Pro Ser Thr Thr 340 345 350Asp Thr Ser Thr Arg Lys Ser Gln Ala Val Asp Ile Asp Tyr Lys Asp 355 360 365Asp Asp Asp Lys Ile Glu Gly Arg Met Asp Gly Ala Arg Ala Ser Val 370 375 380Leu Ser Gly Gly Glu Leu Asp Arg Trp Glu Lys Ile Arg Leu Arg Pro385 390 395 400Gly Gly Lys Lys Lys Tyr Lys Leu Lys His Ile Val Trp Ala Ser Arg 405 410 415Glu Leu Glu Arg Phe Ala Val Asn Pro Gly Leu Leu Glu Thr Ser Glu 420 425 430Gly Cys Arg Gln Ile Leu Gly Gln Leu Gln Pro Ser Leu Gln Thr Gly 435 440 445Ser Glu Glu Leu Arg Ser Leu Tyr Asn Thr Val Ala Thr Leu Tyr Cys 450 455 460Val His Gln Arg Ile Glu Ile Lys Asp Thr Lys Glu Ala Leu Asp Lys465 470 475 480Ile Glu Glu Glu Gln Asn Lys Ser Lys Lys Lys Ala Gln Gln Ala Ala 485 490 495Ala Asp Thr Gly His Ser Ser Gln Val Ser Gln Asn Tyr Pro Ile Val 500 505 510Gln Asn Ile Gln Gly Gln Met Val His Gln Ala Ile Ser Pro Arg Thr 515 520 525Leu Asn Ala Trp Val Lys Val Val Glu Glu Lys Ala Phe Ser Pro Glu 530 535 540Val Ile Pro Met Phe Ser Ala Leu Ser Glu Gly Ala Thr Pro Gln Asp545 550 555 560Leu Asn Thr Met Leu Asn Thr Val Gly Gly His Gln Ala Ala Met Gln 565 570 575Met Leu Lys Glu Thr Ile Asn Glu Glu Ala Ala Glu Trp Asp Arg Val 580 585 590His Pro Val His Ala Gly Pro Ile Ala Pro Gly Gln Met Arg Glu Pro 595 600 605Arg Gly Ser Asp Ile Ala Gly Thr Thr Ser Thr Leu Gln Glu Gln Ile 610 615 620Gly Trp Met Thr Asn Asn Pro Pro Ile Pro Val Gly Glu Ile Tyr Lys625 630 635 640Arg Trp Ile Ile Leu Gly Leu Asn Lys Ile Val Arg Met Tyr Ser Pro 645 650 655Thr Ser Ile Leu Asp Ile Arg Gln Gly Pro Lys Glu Pro Phe Arg Asp 660 665 670Tyr Val Asp Arg Phe Tyr Lys Thr Leu Arg Ala Glu Gln Ala Ser Gln 675 680 685Glu Val Lys Asn Trp Met Thr Glu Thr Leu Leu Val Gln Asn Ala Asn 690 695 700Pro Asp Cys Lys Thr Ile Leu Lys Ala Leu Gly Pro Ala Ala Thr Leu705 710 715 720Glu Glu Met Met Thr Ala Cys Gln Gly Val Gly Gly Pro Gly His Lys 725 730 735Ala Arg Val Leu Ala Glu Ala Met Ser Gln Val Thr Asn Thr Ala Thr 740 745 750Ile Met Met Gln Arg Gly Asn Phe Arg Asn Gln Arg Lys Met Val Lys 755 760 765Cys Phe Asn Cys Gly Lys Glu Gly His Thr Ala Arg Asn Cys Arg Ala 770 775 780Pro Arg Lys Lys Gly Cys Trp Lys Cys Gly Lys Glu Gly His Gln Met785 790 795 800Lys Asp Cys Thr Glu Arg Gln Ala Asn Phe Leu Gly Lys Ile Trp Pro 805 810 815Ser Tyr Lys Gly Arg Pro Gly Asn Phe Leu Gln Ser Arg Pro Glu Pro 820 825 830Thr Ala Pro Pro Phe Leu Gln Ser Arg Pro Glu Pro Thr Ala Pro Pro 835 840 845Glu Glu Ser Phe Arg Ser Gly Val Glu Thr Thr Thr Pro Pro Gln Lys 850 855 860Gln Glu Pro Ile Asp Lys Glu Leu Tyr Pro Leu Thr Ser Leu Arg Ser865 870 875 880Leu Phe Gly Asn Asp Pro Ser Ser Gln Val Asn Ser Arg Gly Leu Asn 885 890 895Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp His Glu 900 905275PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic peptide" 27Ser Tyr Ala Met His1 52819PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic peptide" 28Arg Ile Lys Ser Lys Ala Gln Gly Gly Thr Thr Asp Tyr Ala Ala His1 5 10 15Val Lys Gly298PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic peptide" 29Val Ser Phe Ser Thr Phe Asp Val1 5307PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic peptide" 30Gly Phe Thr Phe Ser Ser Tyr1 5318PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic peptide" 31Lys Ser Lys Ala Gln Gly Gly Thr1 53213PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic peptide" 32Ser Gly Ser Ser Ser Asn Ile Gly Ser Tyr Tyr Val Ser1 5 10337PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic peptide" 33Arg Asn Asn Gln Arg Pro Ser1 53410PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic peptide" 34Asp Ser Trp Asp His Ser Ser Met Asn Val1 5 1035119PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 35Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Gly Arg Ile Lys Ser Lys Ala Gln Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60His Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr65 70 75 80Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95Tyr Cys Ala Arg Val Ser Phe Ser Thr Phe Asp Val Trp Gly Gln Gly 100 105 110Thr Leu Val Thr Val Ser Ser 11536111PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 36Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Tyr 20 25 30Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Val Leu 35 40 45Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln65 70 75 80Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Asp Ser Trp Asp His Ser Ser 85 90 95Met Asn Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 11037449PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 37Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Gly Arg Ile Lys Ser Lys Ala Gln Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60His Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr65 70 75 80Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95Tyr Cys Ala Arg Val Ser Phe Ser Thr Phe Asp Val Trp Gly Gln Gly 100 105 110Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp145 150 155 160Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys 210 215 220Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Glu Gly Ala Pro225 230 235 240Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu305 310 315 320Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys 325 330 335Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu385 390 395 400Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420

425 430Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445Lys38215PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 38Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Tyr 20 25 30Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Val Leu 35 40 45Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln65 70 75 80Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Asp Ser Trp Asp His Ser Ser 85 90 95Met Asn Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro 100 105 110Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu 115 120 125Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro 130 135 140Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala145 150 155 160Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala 165 170 175Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg 180 185 190Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr 195 200 205Val Ala Pro Thr Glu Cys Ser 210 2153919PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic peptide" 39Arg Ile Lys Ser Val Ala Gln Gly Gly Thr Thr Asp Tyr Ala Ala His1 5 10 15Val Lys Gly408PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic peptide" 40Val Ser His Ser Thr Phe Asp Val1 5418PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic peptide" 41Lys Ser Val Ala Gln Gly Gly Thr1 542119PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 42Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Gly Arg Ile Lys Ser Val Ala Gln Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60His Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr65 70 75 80Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95Tyr Cys Ala Arg Val Ser His Ser Thr Phe Asp Val Trp Gly Gln Gly 100 105 110Thr Leu Val Thr Val Ser Ser 11543111PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 43Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Tyr 20 25 30Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Val Leu 35 40 45Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln65 70 75 80Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Asp Ser Trp Asp His Ser Ser 85 90 95Met Asn Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln 100 105 11044449PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 44Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Gly Arg Ile Lys Ser Val Ala Gln Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60His Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr65 70 75 80Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 85 90 95Tyr Cys Ala Arg Val Ser His Ser Thr Phe Asp Val Trp Gly Gln Gly 100 105 110Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp145 150 155 160Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys 210 215 220Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Glu Gly Ala Pro225 230 235 240Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu305 310 315 320Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys 325 330 335Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu385 390 395 400Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445Lys45215PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 45Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Tyr 20 25 30Tyr Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Val Leu 35 40 45Ile Tyr Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln65 70 75 80Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Asp Ser Trp Asp His Ser Ser 85 90 95Met Asn Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro 100 105 110Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu 115 120 125Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro 130 135 140Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala145 150 155 160Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala 165 170 175Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg 180 185 190Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr 195 200 205Val Ala Pro Thr Glu Cys Ser 210 2154615DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 46agctatgcga tgcac 154757DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 47cgtatcaaat ccaaagccca gggcggtacg accgactacg cggcgcacgt gaaaggc 574824DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 48gtttctttct ccactttcga tgtt 244921DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 49ggatttacct tcagcagcta t 215024DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 50aaatccaaag cccagggcgg tacg 245139DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 51agcggcagct cctccaatat tggtagctat tacgtgagc 395221DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 52cgtaataatc aacgtcctag c 215330DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 53gacagctggg atcacagctc catgaatgtt 3054357DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 54gaggtgcaat tggtggaaag cggcggtggc ctggtgaaac caggcggcag cctgcgcctg 60agctgcgccg cctccggatt taccttcagc agctatgcga tgcactgggt gcgccaggcc 120ccgggcaaag gtctcgaatg ggtgggtcgt atcaaatcca aagcccaggg cggtacgacc 180gactacgcgg cgcacgtgaa aggccgcttt accattagcc gcgatgattc gaaaaacacc 240ctgtatctgc aaatgaacag cctgaaaacc gaagatacgg ccgtgtatta ttgcgcgcgt 300gtttctttct ccactttcga tgtttggggc caaggcaccc tggtgactgt ctcgagc 35755333DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 55cagagcgtgc tgacccagcc tcctagcgtg agcggtgcac cgggccagcg cgtgaccatt 60agctgtagcg gcagctcctc caatattggt agctattacg tgagctggta tcagcagctg 120ccgggcacgg cgccgaaagt tctgatctat cgtaataatc aacgtcctag cggcgtgccg 180gatcgcttta gcggatccaa aagcggcacc agcgccagcc tggcgattac cggcctgcaa 240gcagaagatg aagcggatta ttactgcgac agctgggatc acagctccat gaatgttttt 300ggcggcggta ccaagctgac cgtgctgggc cag 333561347DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 56gaggtgcaat tggtggaaag cggcggtggc ctggtgaaac caggcggcag cctgcgcctg 60agctgcgccg cctccggatt taccttcagc agctatgcga tgcactgggt gcgccaggcc 120ccgggcaaag gtctcgaatg ggtgggtcgt atcaaatcca aagcccaggg cggtacgacc 180gactacgcgg cgcacgtgaa aggccgcttt accattagcc gcgatgattc gaaaaacacc 240ctgtatctgc aaatgaacag cctgaaaacc gaagatacgg ccgtgtatta ttgcgcgcgt 300gtttctttct ccactttcga tgtttggggc caaggcaccc tggtgactgt ctcgagcgcg 360tcgaccaaag gccccagcgt gttccctctg gcccccagca gcaagagcac ctctggcgga 420acagccgccc tgggctgcct ggtcaaggac tacttccccg agcccgtgac cgtgtcctgg 480aactctggcg ccctgaccag cggcgtgcac acctttccag ccgtgctcca gagcagcggc 540ctgtacagcc tgagcagcgt cgtgaccgtg cccagcagca gcctgggcac ccagacctac 600atctgcaacg tgaaccacaa gcccagcaac acaaaggtgg acaagcgggt ggaacccaag 660agctgcgaca agacccacac ctgtcccccc tgccctgccc ctgaagcgga gggagccccc 720tccgtgttcc tgttcccccc aaagcctaag gacaccctga tgatcagccg gacccccgaa 780gtgacctgcg tggtggtgga cgtgtcccac gaggaccctg aagtgaagtt taattggtac 840gtggacggcg tggaagtgca caacgccaag accaagccca gagaggaaca gtacaacagc 900acctaccggg tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa cggcaaagag 960tacaagtgca aggtgtccaa caaggccctg ccttcctcca tcgagaaaac catcagcaag 1020gccaaaggcc agccccgcga gccccaggtg tacacactgc cccctagccg ggaagagatg 1080accaagaacc aggtgtccct gacctgcctc gtgaagggct tctaccccag cgacattgcc 1140gtggaatggg agagcaacgg ccagcccgag aacaactaca agaccacccc ccctgtgctg 1200gacagcgacg gctcattctt cctgtacagc aagctgaccg tggacaagag ccggtggcag 1260cagggcaacg tgttcagctg ctccgtgatg cacgaggccc tgcacaacca ctacacccag 1320aagtccctga gcctgagccc cggcaag 134757645DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 57cagagcgtgc tgacccagcc tcctagcgtg agcggtgcac cgggccagcg cgtgaccatt 60agctgtagcg gcagctcctc caatattggt agctattacg tgagctggta tcagcagctg 120ccgggcacgg cgccgaaagt tctgatctat cgtaataatc aacgtcctag cggcgtgccg 180gatcgcttta gcggatccaa aagcggcacc agcgccagcc tggcgattac cggcctgcaa 240gcagaagatg aagcggatta ttactgcgac agctgggatc acagctccat gaatgttttt 300ggcggcggta ccaagctgac cgtgctgggc cagcccaaag ccgcccctag cgtgaccctg 360ttccccccct cgagtgagga actccaggcc aacaaggcca ccctcgtgtg cctgatcagc 420gacttctacc ctggcgccgt gaccgtggcc tggaaggccg atagcagccc tgtgaaggcc 480ggcgtggaaa ccaccacccc cagcaagcag agcaacaaca aatacgccgc cagcagctac 540ctgagcctga cccccgagca gtggaagtcc cacagatcct acagctgcca ggtcacacac 600gagggcagca ccgtggaaaa gaccgtggcc cccaccgagt gcagc 6455815DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 58agctatgcga tgcac 155957DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 59cgtatcaaat ccgtggccca gggcggtacg accgactacg cggcgcacgt gaaaggc 576024DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 60gtttctcatt ccactttcga tgtt 246121DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 61ggatttacct tcagcagcta t 216224DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 62aaatccgtgg cccagggcgg tacg 246339DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 63agcggcagct cctccaatat tggtagctat tacgtgagc 396421DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 64cgtaataatc aacgtcctag c 216530DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 65gacagctggg atcacagctc catgaatgtt 3066357DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 66gaggtgcaat tggtggaaag cggcggtggc ctggtgaaac caggcggcag cctgcgcctg 60agctgcgccg cctccggatt taccttcagc agctatgcga tgcactgggt gcgccaggcc 120ccgggcaaag gtctcgaatg ggtgggtcgt atcaaatccg tggcccaggg cggtacgacc 180gactacgcgg cgcacgtgaa aggccgcttt accattagcc gcgatgattc gaaaaacacc 240ctgtatctgc aaatgaacag cctgaaaacc gaagatacgg ccgtgtatta ttgcgcgcgt 300gtttctcatt ccactttcga tgtttggggc caaggcaccc tggtgactgt ctcgagc 35767333DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 67cagagcgtgc tgacccagcc tcctagcgtg agcggtgcac cgggccagcg cgtgaccatt 60agctgtagcg gcagctcctc caatattggt agctattacg tgagctggta tcagcagctg 120ccgggcacgg cgccgaaagt tctgatctat cgtaataatc aacgtcctag cggcgtgccg 180gatcgcttta gcggatccaa aagcggcacc agcgccagcc tggcgattac cggcctgcaa 240gcagaagatg aagcggatta ttactgcgac agctgggatc acagctccat gaatgttttt 300ggcggcggta ccaagctgac cgtgctgggc cag 333681347DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 68gaggtgcaat tggtggaaag cggcggtggc ctggtgaaac caggcggcag cctgcgcctg 60agctgcgccg cctccggatt taccttcagc agctatgcga tgcactgggt gcgccaggcc 120ccgggcaaag gtctcgaatg ggtgggtcgt atcaaatccg tggcccaggg cggtacgacc 180gactacgcgg cgcacgtgaa aggccgcttt accattagcc gcgatgattc gaaaaacacc 240ctgtatctgc aaatgaacag cctgaaaacc gaagatacgg ccgtgtatta ttgcgcgcgt 300gtttctcatt ccactttcga tgtttggggc caaggcaccc tggtgactgt ctcgagcgcg 360tcgaccaaag gccccagcgt gttccctctg gcccccagca gcaagagcac ctctggcgga 420acagccgccc tgggctgcct ggtcaaggac tacttccccg agcccgtgac cgtgtcctgg 480aactctggcg ccctgaccag cggcgtgcac acctttccag ccgtgctcca

gagcagcggc 540ctgtacagcc tgagcagcgt cgtgaccgtg cccagcagca gcctgggcac ccagacctac 600atctgcaacg tgaaccacaa gcccagcaac acaaaggtgg acaagcgggt ggaacccaag 660agctgcgaca agacccacac ctgtcccccc tgccctgccc ctgaagcgga gggagccccc 720tccgtgttcc tgttcccccc aaagcctaag gacaccctga tgatcagccg gacccccgaa 780gtgacctgcg tggtggtgga cgtgtcccac gaggaccctg aagtgaagtt taattggtac 840gtggacggcg tggaagtgca caacgccaag accaagccca gagaggaaca gtacaacagc 900acctaccggg tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa cggcaaagag 960tacaagtgca aggtgtccaa caaggccctg ccttcctcca tcgagaaaac catcagcaag 1020gccaaaggcc agccccgcga gccccaggtg tacacactgc cccctagccg ggaagagatg 1080accaagaacc aggtgtccct gacctgcctc gtgaagggct tctaccccag cgacattgcc 1140gtggaatggg agagcaacgg ccagcccgag aacaactaca agaccacccc ccctgtgctg 1200gacagcgacg gctcattctt cctgtacagc aagctgaccg tggacaagag ccggtggcag 1260cagggcaacg tgttcagctg ctccgtgatg cacgaggccc tgcacaacca ctacacccag 1320aagtccctga gcctgagccc cggcaag 134769645DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 69cagagcgtgc tgacccagcc tcctagcgtg agcggtgcac cgggccagcg cgtgaccatt 60agctgtagcg gcagctcctc caatattggt agctattacg tgagctggta tcagcagctg 120ccgggcacgg cgccgaaagt tctgatctat cgtaataatc aacgtcctag cggcgtgccg 180gatcgcttta gcggatccaa aagcggcacc agcgccagcc tggcgattac cggcctgcaa 240gcagaagatg aagcggatta ttactgcgac agctgggatc acagctccat gaatgttttt 300ggcggcggta ccaagctgac cgtgctgggc cagcccaaag ccgcccctag cgtgaccctg 360ttccccccct cgagtgagga actccaggcc aacaaggcca ccctcgtgtg cctgatcagc 420gacttctacc ctggcgccgt gaccgtggcc tggaaggccg atagcagccc tgtgaaggcc 480ggcgtggaaa ccaccacccc cagcaagcag agcaacaaca aatacgccgc cagcagctac 540ctgagcctga cccccgagca gtggaagtcc cacagatcct acagctgcca ggtcacacac 600gagggcagca ccgtggaaaa gaccgtggcc cccaccgagt gcagc 6457015DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 70agctacgcta tgcac 157157DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 71cggatcaaga gcaaggctca aggcggcacc accgattacg ccgctcatgt gaagggc 577224DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 72gtgtccttct ccaccttcga tgtg 247321DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 73ggcttcacct tctccagcta c 217424DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 74aagagcaagg ctcaaggcgg cacc 247539DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 75tccggctcct cctccaacat cggctcctac tacgtgtcc 397621DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 76cggaacaacc agcggccttc t 217730DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 77gactcttggg accactcctc catgaacgtg 3078357DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 78gaagtgcagc tggtggaatc tggcggcgga cttgtgaaac ctggcggctc tctgagactg 60tcttgtgccg cttccggctt caccttctcc agctacgcta tgcactgggt ccgacaggcc 120cctggcaaag gattggagtg ggtcggacgg atcaagagca aggctcaagg cggcaccacc 180gattacgccg ctcatgtgaa gggcagattc accatctctc gggacgactc caagaacacc 240ctgtacctgc agatgaactc cctgaaaacc gaggacaccg ccgtgtacta ctgcgccaga 300gtgtccttct ccaccttcga tgtgtggggc cagggcacac tggttacagt ctcgagc 35779333DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 79cagtccgtgc tgacccagcc tccttctgtt tctggtgctc ctggccagag agtgaccatc 60tcttgctccg gctcctcctc caacatcggc tcctactacg tgtcctggta tcagcagctg 120cctggcaccg ctcctaaggt gctgatctac cggaacaacc agcggccttc tggcgtgccc 180gatagattct ccggctctaa gtctggcacc tctgccagcc tggctatcac tggactgcag 240gctgaggacg aggccgacta ctactgcgac tcttgggacc actcctccat gaacgtgttc 300ggcggaggta ccaagctgac cgtgctggga cag 333801347DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 80gaagtgcagc tggtggaatc tggcggcgga cttgtgaaac ctggcggctc tctgagactg 60tcttgtgccg cttccggctt caccttctcc agctacgcta tgcactgggt ccgacaggcc 120cctggcaaag gattggagtg ggtcggacgg atcaagagca aggctcaagg cggcaccacc 180gattacgccg ctcatgtgaa gggcagattc accatctctc gggacgactc caagaacacc 240ctgtacctgc agatgaactc cctgaaaacc gaggacaccg ccgtgtacta ctgcgccaga 300gtgtccttct ccaccttcga tgtgtggggc cagggcacac tggttacagt ctcgagcgcc 360tccaccaaag gaccctctgt gtttcctctg gctccctcca gcaagtctac ctctggtgga 420acagctgccc tgggctgcct ggtcaaggat tactttcctg agcctgtgac cgtgtcctgg 480aactctggcg ctctgacatc tggcgtgcac acctttccag ctgtgctgca gtcctctggc 540ctgtacagcc tgtcctctgt cgtgaccgtg ccttctagct ctctgggcac ccagacctac 600atctgcaatg tgaaccacaa gccttccaac accaaggtgg acaagagagt ggaacccaag 660tcctgcgaca agacccacac ctgtcctcca tgtcctgctc cagaagctga gggcgctcct 720tccgtgttcc tgtttcctcc aaagcctaag gacaccctga tgatctctcg gacccctgaa 780gtgacctgcg tggtggtgga tgtgtctcac gaggacccag aagtgaagtt caattggtac 840gtggacggcg tggaagtgca caacgccaag accaagccta gagaggaaca gtacaactcc 900acctacagag tggtgtccgt gctgaccgtg ctgcaccagg attggctgaa cggcaaagag 960tacaagtgca aggtgtccaa caaggccctg ccttccagca tcgaaaagac catctccaag 1020gccaagggcc agcctaggga accccaggtt tacaccctgc ctccaagccg ggaagagatg 1080accaagaacc aggtgtccct gacctgcctc gtgaagggct tctacccttc cgatatcgcc 1140gtggaatggg agagcaatgg ccagcctgag aacaactaca agacaacccc tcctgtgctg 1200gactccgacg gctcattctt cctgtactcc aagctgacag tggacaagtc cagatggcag 1260cagggcaacg tgttctcctg ctccgtgatg cacgaggccc tgcacaatca ctacacacag 1320aagtccctgt ctctgtcccc tggcaag 134781645DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 81cagtccgtgc tgacccagcc tccttctgtt tctggtgctc ctggccagag agtgaccatc 60tcttgctccg gctcctcctc caacatcggc tcctactacg tgtcctggta tcagcagctg 120cctggcaccg ctcctaaggt gctgatctac cggaacaacc agcggccttc tggcgtgccc 180gatagattct ccggctctaa gtctggcacc tctgccagcc tggctatcac tggactgcag 240gctgaggacg aggccgacta ctactgcgac tcttgggacc actcctccat gaacgtgttc 300ggcggaggta ccaagctgac cgtgctggga cagcctaagg ctgccccttc cgtgacactg 360ttccctccat cctctgagga actgcaggcc aacaaggcta ccctcgtgtg cctgatctcc 420gacttttacc ctggcgctgt gaccgtggcc tggaaggctg atagttctcc tgtgaaggcc 480ggcgtggaaa ccaccacacc ttccaagcag tccaacaaca aatacgccgc ctcctcctac 540ctgtctctga cccctgaaca gtggaagtcc caccggtcct acagctgcca agtgacccat 600gagggctcca ccgtggaaaa gaccgtggct cctaccgagt gctct 6458215DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 82agctacgcta tgcac 158357DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 83cggatcaaga gcgttgccca aggcggcacc accgattacg ctgctcatgt gaagggc 578424DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 84gtgtcccact ctaccttcga tgtg 248521DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 85ggcttcacct tctccagcta c 218624DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 86aagagcgttg cccaaggcgg cacc 248739DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 87tccggctcct cctccaacat cggctcctac tacgtgtcc 398821DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 88cggaacaacc agcggccttc t 218930DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic oligonucleotide" 89gactcttggg accactcctc catgaacgtg 3090357DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 90gaagtgcagc tggtggaatc tggcggcgga cttgtgaaac ctggcggctc tctgagactg 60tcttgtgccg cttccggctt caccttctcc agctacgcta tgcactgggt ccgacaggcc 120cctggcaaag gattggagtg ggtcggacgg atcaagagcg ttgcccaagg cggcaccacc 180gattacgctg ctcatgtgaa gggcagattc accatcagcc gggacgactc caagaacacc 240ctgtacctgc agatgaactc cctgaaaacc gaggacaccg ccgtgtacta ctgcgccaga 300gtgtcccact ctaccttcga tgtgtggggc cagggcacac tggttacagt ctcgagc 35791333DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 91cagtccgtgc tgacccagcc tccttctgtt tctggtgctc ctggccagag agtgaccatc 60tcttgctccg gctcctcctc caacatcggc tcctactacg tgtcctggta tcagcagctg 120cctggcaccg ctcctaaggt gctgatctac cggaacaacc agcggccttc tggcgtgccc 180gatagattct ccggctctaa gtctggcacc tctgccagcc tggctatcac tggactgcag 240gctgaggacg aggccgacta ctactgcgac tcttgggacc actcctccat gaacgtgttc 300ggcggaggta ccaagctgac cgtgctggga cag 333921347DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 92gaagtgcagc tggtggaatc tggcggcgga cttgtgaaac ctggcggctc tctgagactg 60tcttgtgccg cttccggctt caccttctcc agctacgcta tgcactgggt ccgacaggcc 120cctggcaaag gattggagtg ggtcggacgg atcaagagcg ttgcccaagg cggcaccacc 180gattacgctg ctcatgtgaa gggcagattc accatcagcc gggacgactc caagaacacc 240ctgtacctgc agatgaactc cctgaaaacc gaggacaccg ccgtgtacta ctgcgccaga 300gtgtcccact ctaccttcga tgtgtggggc cagggcacac tggttacagt ctcgagcgcc 360tccaccaaag gaccctctgt gtttcctctg gctccctcca gcaagtctac ctctggtgga 420acagctgccc tgggctgcct ggtcaaggat tactttcctg agcctgtgac cgtgtcctgg 480aactctggcg ctctgacatc tggcgtgcac acctttccag ctgtgctgca gtcctctggc 540ctgtacagcc tgtcctctgt cgtgaccgtg ccttctagct ctctgggcac ccagacctac 600atctgcaatg tgaaccacaa gccttccaac accaaggtgg acaagagagt ggaacccaag 660tcctgcgaca agacccacac ctgtcctcca tgtcctgctc cagaagctga gggcgctcct 720tccgtgttcc tgtttcctcc aaagcctaag gacaccctga tgatctctcg gacccctgaa 780gtgacctgcg tggtggtgga tgtgtctcac gaggacccag aagtgaagtt caattggtac 840gtggacggcg tggaagtgca caacgccaag accaagccta gagaggaaca gtacaactcc 900acctacagag tggtgtccgt gctgaccgtg ctgcaccagg attggctgaa cggcaaagag 960tacaagtgca aggtgtccaa caaggccctg ccttccagca tcgaaaagac catctccaag 1020gccaagggcc agcctaggga accccaggtt tacaccctgc ctccaagccg ggaagagatg 1080accaagaacc aggtgtccct gacctgcctc gtgaagggct tctacccttc cgatatcgcc 1140gtggaatggg agagcaatgg ccagcctgag aacaactaca agacaacccc tcctgtgctg 1200gactccgacg gctcattctt cctgtactcc aagctgacag tggacaagtc cagatggcag 1260cagggcaacg tgttctcctg ctccgtgatg cacgaggccc tgcacaatca ctacacacag 1320aagtccctgt ctctgtcccc tggcaag 134793645DNAArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polynucleotide" 93cagtccgtgc tgacccagcc tccttctgtt tctggtgctc ctggccagag agtgaccatc 60tcttgctccg gctcctcctc caacatcggc tcctactacg tgtcctggta tcagcagctg 120cctggcaccg ctcctaaggt gctgatctac cggaacaacc agcggccttc tggcgtgccc 180gatagattct ccggctctaa gtctggcacc tctgccagcc tggctatcac tggactgcag 240gctgaggacg aggccgacta ctactgcgac tcttgggacc actcctccat gaacgtgttc 300ggcggaggta ccaagctgac cgtgctggga cagcctaagg ctgccccttc cgtgacactg 360ttccctccat cctctgagga actgcaggcc aacaaggcta ccctcgtgtg cctgatctcc 420gacttttacc ctggcgctgt gaccgtggcc tggaaggctg atagttctcc tgtgaaggcc 480ggcgtggaaa ccaccacacc ttccaagcag tccaacaaca aatacgccgc ctcctcctac 540ctgtctctga cccctgaaca gtggaagtcc caccggtcct acagctgcca agtgacccat 600gagggctcca ccgtggaaaa gaccgtggct cctaccgagt gctct 64594454PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 94Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Val Met His Trp Val Arg Gln Ala Thr Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Ala Ile Asp Thr Gly Gly Gly Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60Gly Arg Phe Thr Ile Ser Arg Glu Asn Ala Lys Asn Ser Leu Tyr Leu65 70 75 80Gln Met Asn Ser Leu Arg Ala Gly Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Asp Tyr Tyr Tyr Tyr Ala Ser Gly Ser Tyr Tyr Lys Ala Phe Asp 100 105 110Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 130 135 140Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn 195 200 205Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro 210 215 220Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu225 230 235 240Ala Glu Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 245 250 255Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 260 265 270Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 275 280 285Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 290 295 300Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp305 310 315 320Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 325 330 335Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 340 345 350Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 355 360 365Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 370 375 380Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr385 390 395 400Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 405 410 415Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 420 425 430Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 435 440 445Ser Leu Ser Pro Gly Lys 45095214PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 95Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Arg 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Pro Leu 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205Phe Asn Arg Gly Glu Cys 21096400PRTArtificial Sequencesource/note="Description of Artificial Sequence Synthetic polypeptide" 96Ala Glu Pro Arg Leu Pro Leu Met Tyr His Leu Ala Ala Val Ser Asp1 5 10 15Leu Ser Thr Gly Leu Pro Ser Phe Trp Ala Thr Gly Trp Leu Gly Ala 20 25 30Gln Gln Tyr Leu Thr Tyr Asn Asn Leu Arg Gln Glu Ala Asp Pro Cys 35

40 45Gly Ala Trp Ile Trp Glu Asn Gln Val Ser Trp Tyr Trp Glu Lys Glu 50 55 60Thr Thr Asp Leu Lys Ser Lys Glu Gln Leu Phe Leu Glu Ala Ile Arg65 70 75 80Thr Leu Glu Asn Gln Ile Asn Gly Thr Phe Thr Leu Gln Gly Leu Leu 85 90 95Gly Cys Glu Leu Ala Pro Asp Asn Ser Ser Leu Pro Thr Ala Val Phe 100 105 110Ala Leu Asn Gly Glu Glu Phe Met Arg Phe Asn Pro Arg Thr Gly Asn 115 120 125Trp Ser Gly Glu Trp Pro Glu Thr Asp Ile Val Gly Asn Leu Trp Met 130 135 140Lys Gln Pro Glu Ala Ala Arg Lys Glu Ser Glu Phe Leu Leu Thr Ser145 150 155 160Cys Pro Glu Arg Leu Leu Gly His Leu Glu Arg Gly Arg Gln Asn Leu 165 170 175Glu Trp Lys Glu Pro Pro Ser Met Arg Leu Lys Ala Arg Pro Gly Asn 180 185 190Ser Gly Ser Ser Val Leu Thr Cys Ala Ala Phe Ser Phe Tyr Pro Pro 195 200 205Glu Leu Lys Phe Arg Phe Leu Arg Asn Gly Leu Ala Ser Gly Ser Gly 210 215 220Asn Cys Ser Thr Gly Pro Asn Gly Asp Gly Ser Phe His Ala Trp Ser225 230 235 240Leu Leu Glu Val Lys Arg Gly Asp Glu His His Tyr Gln Cys Gln Val 245 250 255Glu His Glu Gly Leu Ala Gln Pro Leu Thr Val Asp Leu Asp Ser Pro 260 265 270Ala Arg Ser Ser Val Asn Ser Arg Gly Leu Asn Asp Ile Phe Glu Ala 275 280 285Gln Lys Ile Glu Trp His Glu His His His His His His Ile Gln Lys 290 295 300Thr Pro Gln Ile Gln Val Tyr Ser Arg His Pro Pro Glu Asn Gly Lys305 310 315 320Pro Asn Phe Leu Asn Cys Tyr Val Ser Gln Phe His Pro Pro Gln Ile 325 330 335Glu Ile Glu Leu Leu Lys Asn Gly Lys Lys Ile Pro Asn Ile Glu Met 340 345 350Ser Asp Leu Ser Phe Ser Lys Asp Trp Ser Phe Tyr Ile Leu Ala His 355 360 365Thr Glu Phe Thr Pro Thr Glu Thr Asp Val Tyr Ala Cys Arg Val Lys 370 375 380His Val Thr Leu Lys Glu Pro Lys Thr Val Thr Trp Asp Arg Asp Met385 390 395 400

Claims

1: An isolated antibody or antibody fragment specific for human C5aR, wherein said antibody or antibody fragment comprises a) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 28, the HCDR3 region of SEQ ID NO: 29, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34, or b) the HCDR1 region of SEQ ID NO: 27, the HCDR2 region of SEQ ID NO: 39, the HCDR3 region of SEQ ID NO: 40, the LCDR1 region of SEQ ID NO: 32, the LCDR2 region of SEQ ID NO: 33 and the LCDR3 region of SEQ ID NO: 34.

2: The isolated antibody or antibody fragment according to claim 1, wherein said antibody or antibody fragment comprises a) the VH of SEQ ID NO: 35 and the VL of SEQ ID NO 36, or b) the VH of SEQ ID NO: 42 and the VL of SEQ ID NO 43.

3: The isolated antibody or antibody fragment according to claim 1, wherein said antibody or antibody fragment comprises a) the HC of SEQ ID NO: 37 and the LC of SEQ ID NO 38, or b) the HC of SEQ ID NO: 44 and the LC of SEQ ID NO 45.

4: The isolated antibody or antibody fragment according to claim 1, wherein said antibody or antibody fragment is of the human IgG1 class.

5: The isolated antibody or antibody fragment according to claim 1, wherein said antibody or antibody fragment does not substantially induce effector function in vitro.

6: The isolated antibody or antibody fragment according to claim 1, wherein said antibody or antibody fragment comprises one or more amino acid substitution selected from the group of: L234A, L235E, G237A, A330S and P331S, with numbering according to EU index.

7: The isolated antibody or antibody fragment according to claim 1, wherein said isolated antibody or antibody fragment is specific for human C5aR and cynomolgus C5aR.

8: The isolated antibody or antibody fragment according to claim 1, wherein said isolated antibody or antibody fragment inhibits human C5a induced CD11b expression in human granulocytes with an IC.sub.50 concentration of 42 nM in the presence of 150 nM human C5a in vitro.

9: The isolated antibody or antibody fragment according to claim 1, wherein said isolated antibody or antibody fragment is a monoclonal antibody or antibody fragment.

10: The isolated antibody or antibody fragment according to claim 1, wherein said isolated antibody or antibody fragment is a human, humanized or chimeric antibody or antibody fragment.

11: The A method for altering biological activity of human C5aR comprising administering the isolated antibody or antibody fragment of claim 1.

12: A nucleic acid composition comprising a nucleic acid sequence or a plurality of nucleic acid sequences encoding the isolated antibody or antibody fragment according to claim 1.

13: A vector composition comprising a vector or a plurality of vectors comprising the nucleic acid composition of claim 12.

14: A host cell comprising the vector composition of claim 13.

15: A pharmaceutical composition comprising the isolated antibody or antibody fragment according to claim 1 and a pharmaceutically acceptable carrier or excipient.

16: A host cell comprising the nucleic acid composition of claim 12.

17: A method for treating a disease in a subject comprising administering to the subject the antibody or antibody fragment of claim 1.

18: A method for detecting C5aR in a subject or a sample, said method comprising the step of contacting said subject or sample with the antibody or antibody fragment of claim 1.