CN114149501A

CN114149501A - anti-C5 antibodies and uses thereof

Info

Publication number: CN114149501A
Application number: CN202111444150.8A
Authority: CN
Inventors: 吕明启; 李剑; 黄瑞晶; 于永生; 王根辈; 王轶博; 李文蕾
Original assignee: Tasly Pharmaceutical Group Co Ltd
Current assignee: Tasly Pharmaceutical Group Co Ltd
Priority date: 2020-12-11
Filing date: 2021-11-30
Publication date: 2022-03-08
Anticipated expiration: 2041-11-30
Also published as: CN114149501B; WO2022122020A1

Abstract

The invention provides an anti-C5 antibody and application thereof, wherein a variable region of the anti-C5 antibody comprises an amino acid sequence shown in SEQ ID NO. 19-20, SEQ ID NO. 21-22 or SEQ ID NO. 23-24. The anti-C5 antibodies CR3#24, CR3#30 and CR3#75, especially CR3#75 have obvious binding capacity with humanized recombinant C5 protein, show obvious CDC inhibition effect, have better binding activity compared with Eculizumab and C5, have long half-life, good solubility and good thermal stability, and have potential application value in the aspect of pharmacy development.

Description

anti-C5 antibodies and uses thereof

The present application claims priority from the application serial No. 202011459619.0 (prior application filed 12/11/2020 entitled anti-C5 antibody and uses thereof).

Technical Field

The invention belongs to the technical field of biological medicines, and relates to an anti-C5 antibody and application thereof.

Background

The complement system consists of more than 30 plasma proteins and membrane proteins, is widely present on the surfaces of blood, tissue fluid and cells, promotes the phagocytosis of phagocytes and the capability of dissolving target cells, and is an important component of the immune defense mechanism of the body. Among them, C5 is an important complement, C5 convertase breaks C5 into C5a and C5b during inflammatory reaction, C5a is free in liquid phase and is an important inflammatory mediator, and C5b forms a Membrane Attack Complex (MAC) through a series of complement reactions, resulting in cell lysis death.

Normally complement activation and inhibition mechanisms work together to maintain a healthy body state, and when C5 is over-activated due to some conditions, abnormal autoimmunity is generated. For example, Paroxysmal Nocturnal Hemoglobinuria (PNH) is caused by impaired C5 inhibition due to a deficiency in CD59 of PNH red blood cells, and atypical hemolytic uremic syndrome (aHUS) is caused by excessive activation of the alternative complement pathway leading to the formation of membrane attack complexes and damage to microvascular endothelial cells, all of which share a common mechanism of action of C5 over-activation.

Disclosure of Invention

The anti-C5 antibody can be combined with C5 with high affinity, so that the C5 is prevented from being cracked by C5 invertase, the formation of a membrane attack blocking complex is facilitated, and the aim of relieving and improving clinical symptoms is fulfilled finally.

In order to achieve the purpose, the invention adopts the following technical scheme:

in a first aspect, the present invention provides an anti-C5 antibody, said anti-C5 antibody comprising a heavy chain variable region and a light chain variable region;

the heavy chain variable region comprises CDR3 shown in SEQ ID NO. 3, SEQ ID NO. 9 or SEQ ID NO. 15;

the light chain variable region includes CDR3 shown in SEQ ID NO 6, SEQ ID NO 12 or SEQ ID NO 18.

Preferably, the heavy chain variable region further comprises CDR1 shown in SEQ ID NO. 1, SEQ ID NO. 7 or SEQ ID NO. 13.

Preferably, the heavy chain variable region further comprises CDR2 shown in SEQ ID NO 2, SEQ ID NO 8 or SEQ ID NO 14.

Preferably, the light chain variable region further comprises CDR1 shown in SEQ ID NO. 4, SEQ ID NO. 10 or SEQ ID NO. 16.

Preferably, the light chain variable region further comprises CDR2 shown in SEQ ID NO 5, SEQ ID NO 11 or SEQ ID NO 17.

In the invention, CDRs 1-3 of a heavy chain variable region and CDRs 1-3 of a light chain variable region of an antibody jointly determine the specific recognition and binding capacity of the antibody to an antigen, and the antibody containing the CDRs of SEQ ID NO 1-6, SEQ ID NO 7-12 and SEQ ID NO 13-18 has remarkable binding capacity to C5 protein and shows obvious CDC inhibition effect.

In a specific embodiment, the heavy chain variable region of the anti-C5 antibody CR3#24 comprises the CDR1 of SEQ ID NO:1, the CDR2 of SEQ ID NO:2, the CDR3 of SEQ ID NO: 3;

the variable region of the light chain of the anti-C5 antibody CR3#24 comprises CDR1 shown in SEQ ID NO. 4, CDR2 shown in SEQ ID NO. 5 and CDR3 shown in SEQ ID NO. 6;

SEQ ID NO:1：DYYMH；

SEQ ID NO:2：VVDPEDGEIIYAEKFQG；

SEQ ID NO:3：SDYGAGSYMVDP；

SEQ ID NO:4：GGNNLESKIVS；

SEQ ID NO:5：YDTDRPS；

SEQ ID NO:6：QVWDSNSDHVI。

in the invention, the anti-C5 antibody CR3#24 comprising the heavy chain variable region CDRs of SEQ ID NOS: 1-3 and the light chain variable region CDRs of SEQ ID NOS: 4-6 has a C5 protein binding activity and shows an obvious CDC inhibitory action.

Preferably, the heavy chain variable region of the anti-C5 antibody CR #24 comprises the amino acid sequence shown in SEQ ID NO. 19, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO. 20;

SEQ ID NO:19：

EVQLVQSGAEVKKPGAAVKISCKVSGYTFTDYYMHWVQQAPGKGLEWMGVVDPEDGEIIYAEKFQGRITITADTSTDTVYMELSSLRSEDTAVYYCARSDYGAGSYMVDPWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK；

SEQ ID NO:20：

SYVLTQPPSVSVTPGKTARIACGGNNLESKIVSWYQQKPGQAPVLVIFYDTDRPSGISERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSNSDHVIFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLVSDFYPGAVTVAWKADGSPVKVGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCRVTHEGSTVEKTVATAECS。

in a specific embodiment, the heavy chain variable region of the anti-C5 antibody CR3#30 comprises the CDR1 of SEQ ID NO:7, the CDR2 of SEQ ID NO:8, the CDR3 of SEQ ID NO: 9;

the variable region of the light chain of the anti-C5 antibody CR3#30 comprises CDR1 shown in SEQ ID NO. 10, CDR2 shown in SEQ ID NO. 11 and CDR3 shown in SEQ ID NO. 12;

SEQ ID NO:7：SYYMH；

SEQ ID NO:8：IINPSGGSTSYAQKFQG；

SEQ ID NO:9：GTVDDAFDI；

SEQ ID NO:10：TGTSSDVGGYDLVS；

SEQ ID NO:11：DVSKRPS；

SEQ ID NO:12：SSFADSNNWEVV。

in the invention, the anti-C5 antibody CR3#30, which contains the heavy chain variable region CDRs of SEQ ID NO. 7-9 and the light chain variable region CDRs of SEQ ID NO. 10-12, has the protein binding activity of C5 and shows an obvious CDC (CDC inhibition) effect.

Preferably, the heavy chain variable region of the anti-C5 antibody CR3#30 comprises the amino acid sequence shown in SEQ ID NO. 21, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO. 22;

SEQ ID NO:21：

QVQLVESGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGTVDDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK；

SEQ ID NO:22：

QSALTQPPSASGSPGQSVTISCTGTSSDVGGYDLVSWYQQHPGKAPKLMIYDVSKRPSGVPDRFSGSKSGNTASLTVSGLQADDEADYYCSSFADSNNWEVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWESHRSYSCQVTHEGSTVEKTVAPTECS。

in a specific embodiment, the heavy chain variable region of the anti-C5 antibody CR3#75 comprises the CDR1 of SEQ ID NO. 13, the CDR2 of SEQ ID NO. 14, the CDR3 of SEQ ID NO. 15;

the variable region of the light chain of the anti-C5 antibody CR3#75 comprises CDR1 shown in SEQ ID NO:16, CDR2 shown in SEQ ID NO:17 and CDR3 shown in SEQ ID NO: 18.

SEQ ID NO:13：DYYTH；

SEQ ID NO:14：LVDPEGGETIYAEKFQG；

SEQ ID NO:15：GSFLAAPDY；

SEQ ID NO:16：KSSQSLLYNSNNKNYLA；

SEQ ID NO:17：WASTRES；

SEQ ID NO:18：QQYYSTPLT。

In the present invention, the anti-C5 antibody CR3#75 comprising the heavy chain variable region CDRs of SEQ ID NOS: 13-15 and the light chain variable region CDRs of SEQ ID NOS: 16-18 has a superior binding activity to that of Eculizumab (Soliris/Alexion) and C5, shows a very strong CDC inhibitory action, has a long half-life, can be retained for a long time, and can achieve a therapeutic purpose by reducing the frequency of administration and the amount of administration.

Preferably, the heavy chain variable region of the anti-C5 antibody CR3#75 comprises the amino acid sequence shown in SEQ ID NO. 23, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO. 24;

SEQ ID NO:23：

EVQLVQSGAEVKKPGATVKISCKVSGYTFTDYYTHWVQQAPGKGLEWMGLVDPEGGETIYAEKFQGRVTITADASTDTAYMELSSLRSEDTAVYYCATGSFLAAPDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK；

SEQ ID NO:24：

DIVMTQAPDSLAVSLGERATVNCKSSQSLLYNSNNKNYLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYFCQQYYSTPLTLGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC。

preferably, the anti-C5 antibody further comprises a constant region.

Preferably, the constant region is derived from any one of IgG4, IgG1 or IgG2, preferably IgG 4.

Preferably, the IgG4 has the S228P mutation, which helps to improve the stability of IgG 4.

Preferably, the IgG1 has the N297A and/or K322A mutation.

Preferably, the IgG2 has the S257A mutation.

In the present invention, the mutation sites N297A and K322A on IgG1 were used to abrogate antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), respectively.

Preferably, the IgG2 has low antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), and may have the S257A mutation.

Preferably, the anti-C5 antibody molecule can be modified at the N-, internal or C-terminus by oligomerization, glycosylation or conjugation to a label, thereby modulating the function of the antibody.

In a second aspect, the present invention provides a nucleic acid molecule comprising a DNA fragment encoding the anti-C5 antibody of the first aspect.

In a third aspect, the present invention provides an expression vector comprising the nucleic acid molecule of the second aspect.

In a fourth aspect, the present invention provides a recombinant cell expressing the anti-C5 antibody of the first aspect.

Preferably, the recombinant cell has integrated into its genome the nucleic acid molecule of the second aspect.

Preferably, the recombinant cell comprises the expression vector of the third aspect.

In a fifth aspect, the present invention provides a method for preparing the anti-C5 antibody of the first aspect, the method comprising the steps of:

(1) connecting the coding nucleic acid of the anti-C5 antibody into a plasmid, transferring the plasmid into a competent cell, and selecting a monoclonal cell for screening after culturing;

(2) extracting the expression vector of the screened positive clone, transferring the expression vector into host cells, culturing, collecting supernatant, and separating and purifying to obtain the antibody.

In a sixth aspect, the present invention provides a pharmaceutical composition comprising the anti-C5 antibody of the first aspect.

Preferably, the pharmaceutical composition further comprises an anti-tumor drug.

Preferably, the pharmaceutical composition further comprises any one or a combination of at least two of a pharmaceutically acceptable carrier, diluent or excipient.

In a seventh aspect, the present invention provides the use of the anti-C5 antibody of the first aspect, the nucleic acid molecule of the second aspect, the expression vector of the third aspect, the recombinant cell of the fourth aspect, or the pharmaceutical composition of the sixth aspect, in the preparation of a disease detection reagent and/or a disease treatment drug.

Preferably, the disease comprises an autoimmune disease resulting from excessive activation of C5.

Compared with the prior art, the invention has the following beneficial effects:

(1) the anti-C5 antibodies CR3#24, CR3#30 and CR3#75, especially CR3#75 have obvious binding capacity with human recombinant C5 protein and show obvious CDC inhibition, and the dissociation equilibrium constant KD of CR3#75 is 0.366-0.722 nM, so that the antibody has better binding activity compared with a control antibody Eculizumab and C5;

(2) the anti-C5 antibody CR3#75 shows excellent pharmacokinetics in a mouse model, and the half life period is as long as 15 days;

(3) the CR3#75/IgG4 monoclonal antibody has a stable structure, an isoelectric point of 6.0-6.9, good solubility and good thermal stability at a low concentration level (>1mg/mL), and a Tm value of 69.83 ℃; compared with CR3#75/IgG4, CR3#75/IgG1(N297A/K322A) has better thermal stability;

(4) the anti-C5 antibody has potential application value in the aspect of preventing and/or treating autoimmune diseases caused by excessive activation of C5.

Drawings

FIG. 1 shows the results of ELISA affinity detection of 128 pieces of C5 specific binding peptide fragments phase obtained by 3 rounds of screening;

FIG. 2 is an SDS-PAGE electrophoresis of C5-specific antibodies containing IgG constant regions, in which M-protein molecular weight Marker, 1-CR3#24, 2-CR3#30, 3-C5 Ref, 4-CR3#25, 5-CR3#29, 6-CR3#37, 7-CR3#47, 8-CR3#53, 9-CR3#56, 10-CR3#74, 11-CR3#75, 12-CR3#79, 13-CR3#110, 14-CR3#112, 15-CR3# 115;

FIG. 3 shows the results of the detection of the activity of the screened C5-specific antibody binding to C5 protein;

FIG. 4A shows the results of CDC inhibitory effect of the screened C5-specific antibody, and FIG. 4B shows the results of CDC inhibitory effect of different concentrations of the C5-specific antibody;

FIG. 5A is the dissociation curve for Eculizumab, and FIG. 5B is the dissociation curve for CR3# 75;

FIG. 6 is a graph of CR3#75/IgG4 metabolism in mice;

FIG. 7 shows the results of an integrity analysis of CR3#75/IgG4, in which panel I is under non-reducing conditions and panel II is under reducing conditions;

FIG. 8 shows the results of isoelectric point detection of CR3#75/IgG4, in which M-protein molecular weight Marker, 1-CR3#75/IgG4 monoclonal antibody;

FIG. 9 shows the results of SEC-HPLC analysis of low concentration CR3#75/IgG4(>1 mg/mL);

FIG. 10 shows the results of thermal stability analysis of CR3#75/IgG 4;

FIG. 11 shows the SEC-HPLC analysis of high concentration CR3#75/IgG4(>10 mg/mL);

fig. 12A is a statistical picture of AQP4 lesion deletion area in mouse brain tissue, and fig. 12B is a statistical picture of GFAP lesion deletion area in mouse brain tissue.

Detailed Description

To further illustrate the technical means adopted by the present invention and the effects thereof, the present invention is further described below with reference to the embodiments and the accompanying drawings. It is to be understood that the specific embodiments described herein are merely illustrative of the invention and are not limiting of the invention.

The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or apparatus used are conventional products commercially available from normal sources, not indicated by the manufacturer.

EXAMPLE 1 screening of active peptide fragments of C5 antibody

In order to obtain a C5 antibody having a therapeutic effect, the present example first screened a peptide fragment having an activity of specifically binding to C5 from the phage antibody library Omnimab. B cells from hundreds of healthy donors were collected from the phage antibody library OmniMab (AP Biosciences Inc.) and displayed based on hyperbage (m 13K07 Δ ρ Ι Ι, Progen, Heidelberg, Germany). Full-length C5 was referenced to NCBI Reference Sequence NP-001726.2 antigen and recombinantly expressed using HEK293 cells (Sino Biologica, Cat #13416-H18H) for enrichment and isolation of specifically binding peptide fragments from the Omnimab pool and HEK293 cells not expressing C5 as controls.

After 3 rounds of screening (CR3) and enrichment, C5 specific binding peptide fragments were identified and isolated using ELISA. As shown in fig. 1, based on clone-specific recognition of C5 activity, CR3#1 to CR3#128 were subjected to sequencing analysis, the sequence information and diversity of heavy and light chains were confirmed, and 15 candidate samples were selected therefrom for subsequent screening. Wherein, the heavy chain variable region and the light chain variable region of CR3#24 are shown as SEQ ID NO 19-20, the heavy chain variable region and the light chain variable region of CR3#30 are shown as SEQ ID NO 21-22, and the heavy chain variable region and the light chain variable region of CR3#75 are shown as SEQ ID NO 23-24.

EXAMPLE 2 expression and purification of C5-specific antibodies containing IgG constant regions

To further evaluate the physicochemical properties of the C5-specific binding peptide fragments after forming intact antibodies, the heavy and light chains of the 15 samples obtained in example 1 were subcloned into an IgG expression vector (AP Biosciences Inc.) carrying the constant region of IgG4(S228P), wherein the mutation site on IgG4 was used to increase the stability of IgG 4.

After the recombinant plasmid is identified by sequencing as a positive clone into which the target fragment is correctly inserted, a sufficient amount of sterile recombinant plasmid is prepared by using a plasmid extraction kit, and transfected to 6X 10 by using a transfection reagent (Invitrogen)⁶Antibody expression was performed in expihho. Recombinant ExpicHO cells at 37 deg.C, 5% CO₂Culturing for 6 days, secreting antibody into serum-free cell culture supernatant, affinity purifying antibody from cell culture supernatant by Protein A chromatography (Protein A chromatography), concentrating, lysing in DPBS buffer solution, detecting Protein concentration by NanoDrop2000, SDS-PAGE, non-reducing, and reducingPurity and integrity tests were performed under reducing conditions, with a protein loading of 5 μ g/lane. After Coomassie blue staining of the gel, ddH was used₂And O, cleaning. Meanwhile, a coulizumab (ecilizumab, Soliris/Alexion) control group (C5 Ref) was set.

As shown in FIG. 2, the integrity and purity of the C5 antibody secreted by ExpicHO cells reach more than 95%, and almost no visible fragments exist in SDS-PAGE gel, which indicates that the steps of screening, subcloning, expressing and purifying the antibody do not influence the structural stability of the antibody protein.

Example 3 detection of the binding Activity of C5-specific antibodies based on direct ELISA

This example examined the binding activity of a C5-specific antibody to a human recombinant C5 protein using a direct ELISA method, and set up an Eculizumab (Soliris/Alexion) control group (C5 Ref). The method comprises the following steps:

human C5 protein (Sino Biologica, Cat #13416-H18H) was diluted to 1. mu.g/mL with 1 XDPBS, added to the ELISA plate at 100. mu.L/well, incubated overnight at 4 ℃ and coated; discarding the coating solution, adding 350 μ L of 1% BSA blocking solution into each well, and blocking at room temperature for 1 h;

preparing a gradient diluted antibody solution to be detected (diluting 3 times from 30nM, diluting 10 times, adding a solution without antibody, and detecting 12 concentrations), adding 100 mu L of antibody solution to be detected with different concentrations into each well of the prepared ELISA plate, and incubating for 1h under room temperature oscillation; discarding the supernatant, adding 350. mu.L/well of washing solution (0.05% Tween-20+1 XDPBS), and washing for 3 times;

adding 100 mu L/well of HRP labeled antibody anti-Fc-Ab-HRP (0.4 mu g/mL) (Jackson immune Research, cat # 109-; discarding the supernatant, adding 350 μ L/hole washing solution, and washing for 3 times;

adding TMB for developing for 1-5 min, adding 50 μ L of stop solution (1N HCl) to stop the reaction, and placing on an enzyme-linked immunosorbent assay (ELISA) reader to read the light absorption value at 450 nm.

As a result, as shown in fig. 3, CR3#75 was screened to have activity of binding C5 protein very close to that of C5 Ref, and CR3#24, CR3#30, and CR3#53 also had significant C5 protein binding activity.

Example 4 functional validation of C5-specific antibodies based on CDC inhibition experiments

The C5-specific antibody was able to block CDC activation, and this example examined the function of the C5-specific antibody based on CDC experiments.

The method comprises the following steps:

to 2X 10⁴10 μ g/mL Rituximab (Rituximab) was added to each Raji cell, incubated at room temperature for 20min, followed by 20 μ g/mL anti-C5 antibody and complement-activated human serum (12.5%) to a final volume of 200 μ L;

incubation was carried out at 37 ℃ for 2h, dead cells were stained with propidium iodide (propidium iodide), and Flow detection was carried out (Attune NxT Flow Cytometer system, thermo fisher scientfic).

As a result, as shown in fig. 4A and 4B, CR3#24, CR3#30, and CR3#75 showed significant CDC inhibitory effects, in which CR3#75 had the strongest neutralizing activity, comparable to that of the control antibody (C5 Ref, Soliris).

Example 5 affinity detection of C5-specific antibodies to human recombinant C5 protein

This example adopts

(Menlo Park, Calif.) the biosensor analyzed the affinity of CR3#75 and C5 and set up an Eculizumab (Eculizumab) control group (C5 Ref). The method comprises the following steps:

briefly, antibodies were loaded onto AHC (Anti-Human IgG Fc Capture) biosensors (Cat #18-5060) at a concentration of 5 μ g/mL, the sensors were equilibrated offline in assay buffer, monitored online until the signal reached about 0.5 nm;

antibody-loaded sensors were exposed to different concentrations (0.4688nM, 0.9375nM, 1.88nM, 3.75nM, 7.5nM, 15nM and 30nM) of C5(Sino Biologica, cat #13416-H18H), followed by 1min incubation in PBST to determine baseline, 5min incubation in C5 solutions of different concentrations to detect the rate of binding of antibody to C5, and 5min incubation in PBST to detect the rate of dissociation of antibody from C5. Kinetic parameters were analyzed using Octet Data software.

As shown in Table 1, Table 2, FIG. 5A and FIG. 5B, CR3#75 has superior binding activity to C5 and a dissociation equilibrium constant KD of 0.366 to 0.722nM compared to the control antibody Eculizumab (KD ═ 0.371 to 0.553 nM).

TABLE 1 binding of Eculizumab to C5

Numbering	KD(M)	kon(1/Ms)	koff(1/s)
				1	5.53E-10	5.44E+05	3.00E-04
2	3.77E-10	3.94E+05	1.49E-04
				3	3.99E-10	3.81E+05	1.52E-04
4	4.35E-10	3.77E+05	1.64E-04
				5	3.71E-10	3.51E+05	1.30E-04

TABLE 2 binding force of CR3#75 to C5

Numbering	KD(M)	kon(1/Ms)	koff(1/s)
				1	3.66E-10	4.13E+05	1.51E-04
2	6.14E-10	4.88E+05	3.00E-04
				3	6.16E-10	4.52E+05	2.79E-04
4	7.22E-10	4.25E+05	3.07E-04
				5	7.20E-10	4.29E+05	3.09E-04

Example 6 pharmacokinetic assays

This example evaluates the half-life of the C5-specific antibody CR3#75/IgG4 in mice, AUC (area under the curve), Cmax, Cl (clearance) and Vd (volume of distribution).

SCID-Bg mice (CB.17/. Cg-PrkdcsccidLystbg-J/CrlBltw) were injected intravenously with 5mg/kg of CR3#75/IgG4 monoclonal antibody and blood samples were collected from the tail vein at various time points. The serum concentration of CR3#75 was measured by direct ELISA and pharmacokinetic parameters were calculated using PKSolver (ver.2.0).

As shown in table 3, table 4 and fig. 6, CR3#75 showed excellent pharmacokinetics in a mouse model, with a half-life of 361.5h (equivalent to 15 days), indicating that the antibody is metabolized slowly in the mouse and can be retained for a long time, reducing the administration frequency and achieving the therapeutic purpose by administering a low dose of the antibody in animal experiments.

TABLE 3

Time (h)	Concentration (μ g/mL)
		0.5	141.692
24	59.527
		72	53.084
168	38.222
		240	42.075
336	28.307
		504	12.811
672	23.378

TABLE 4

Parameter(s)	Unit of	Numerical value
			t1/2	h	361.451
Cmax	μg/mL	141.69
			AUC 0-t	μg/mL×h	22283.99
Vz_obs	(mg/kg)/(μg/mL)	0.0756
			Cl_obs	(mg/kg)/(μg/mL)/h	0.000145
Vss_obs	(mg/kg)/(μg/mL)	0.0838

Example 7 integrity analysis of CR3#75/IgG4 monoclonal antibody

This example uses LabChip GXII to test the integrity of the purified CR3#75/IgG4 monoclonal antibody. The method comprises the following steps:

the purified CR3#75/IgG4 monoclonal antibody was diluted to 1mg/mL with DPBS, 2.5. mu.L of the protein sample dilution was pipetted and mixed with 18. mu.L of sample buffer (containing 1M DTT or 250mM NEM), incubated at 70 ℃ for 10min, and then cooled to room temperature. The system was loaded to a LabChip gxi (Perkin Elmer, inc., cat. cls138160) after adding 35 μ L of deionized water for integrity analysis.

As shown in Table 5 and FIG. 7, the intact CR3#75/IgG4 monoclonal antibody (M.W.172.75kDa) was detected under non-reducing conditions, and the purity of the intact CR3#75/IgG4 monoclonal antibody was found to be 100% by Protein A chromatography (Protein A chromatography), indicating that the heavy chain and the light chain of the antibody are correctly paired, and the antibody can maintain the structural stability during the preparation and purification processes. Under reducing conditions, the sum of heavy and light chains (HC + LC) also reached 96.67%, indicating that most of the impurities were removed using a single purification step.

TABLE 5

EXAMPLE 8 isoelectric Point detection

The present example utilizes IEF gel electrophoresis to detect the charge of the CR3#75/IgG4 monoclonal antibody. The method comprises the following steps:

mu.L of protein sample and 5. mu.L of 2 Xsample buffer were mixed, and 200mL of 1 XIEF cathode buffer or 600mL of 1 XIEF anode buffer were filled in the upper and lower buffer chambers of the electrophoresis apparatus XCell SureLockTMMini-cell (Invitrogen), respectively, under electrophoresis conditions of 1h at 100V, 1h at 200V, and 30min at 500V. After the electrophoresis is finished, the gel is fixed by 12% TCA for 30min, and gel imaging is carried out after Coomassie brilliant blue staining.

As shown in FIG. 8, the CR3#75/IgG4 showed non-uniform charge distribution, and the isoelectric point measured by IEF gel electrophoresis was 6.0 to 6.9, and the predicted isoelectric point was 5.92.

Example 9 aggregative Property analysis

CR3#75/IgG4 antibody was purified and concentrated for SEC-HPLC analysis. The method comprises the following steps:

a concentrated Protein sample (>1mg/mL) was mixed with an equal volume of mobile phase buffer (25mM phosphate, 200mM NaCl, pH 6.8), filtered through a 0.22 μm filter (Millipore, Cat # SLGP003RB), applied to an Xbridge Protein BEH SEC Column (Waters, Cat #186007640) and subjected to SEC separation at 25 ℃ setting a flow rate of 0.4mL/min, sample addition volume of 10 μ L, and absorbance was measured at 280 nm. And finally, data analysis is carried out by adopting Empower 2.

As shown in FIG. 9, the main peak at 17.839min showed that the protein sample had a purity of 99.9% and a concentration of 1.014mg/mL, indicating that CR3#75/IgG4 maintained good solubility in DPBS and substantially no aggregation and precipitation occurred.

Example 10 thermal stability analysis

This example uses Differential Scanning Calorimetry (DSC) to measure the thermal stability of antibodies. The method comprises the following steps:

preparing a protein sample with the concentration of 0.2-1.0 mg/mL, performing DSC detection by using MicroCal PEAQ-DSC system (Northampton, Malvern) at the speed of 200 ℃/h from 10-100 ℃, drawing a thermal analysis diagram, and analyzing the thermal stability of the antibody according to the drawn thermal analysis diagram.

FIG. 10 shows the melting curve of CR3#75/IgG4 with a Tm of 69.83 ℃, indicating that the thermal stability of the antibody is normal.

Example 11 solubility analysis

a concentrated Protein sample (>10mg/mL) was mixed with an equal volume of mobile phase buffer (25mM phosphate, 200mM NaCl, pH 6.8), filtered through a 0.22 μm filter (Millipore, Cat # SLGP003RB), applied to an Xbridge Protein BEH SEC Column (Waters, Cat #186007640) and subjected to SEC separation at 25 ℃ setting a flow rate of 0.4mL/min, sample addition volume of 2 μ L, and absorbance was measured at 280 nm. And finally, data analysis is carried out by adopting Empower 2.

As shown in FIG. 11, the CR3#75/IgG4 antibody could be concentrated to 10mg/mL without macroscopic precipitation, 97.15% protein produced a single peak at 18.005min, and only < 4% protein precipitated, indicating that the antibody was soluble normally and could be further developed into a pharmaceutical.

Example 12 in vivo efficacy

This example demonstrates the in vivo potency of the CR3#75/IgG4 antibody in a NMO mouse model, as follows:

c57 female mice of 6-8 weeks old are anesthetized with chloral hydrate and then are preserved, the heads of the mice are fixed on a brain stereotaxic instrument, holes are drilled after the positioning is adjusted, a micro-sampling needle is vertically inserted into the brain tissue of the mice, an injection pump is started, mixed solution of CR3#75/IgG4 antibody, complement and pathogenic antibody is injected, the needle is pulled out after the injection is finished, the skin is sutured, and the mice are placed in a cage for feeding. Taking out brain tissue after 7 days of modeling to prepare frozen tissue sections, staining AQP4 and GFAP expressed by astrocytes by adopting an immunofluorescence technique, and counting the lesion deletion areas of AQP4 and GFAP of each group.

As a result, as shown in FIGS. 12A and 12B, the CR3#75/IgG4 antibody inhibited the formation of NMO-typical lesions at concentrations of 0.04mg/mL, 0.156mg/mL, 0.625mg/mL, and 2.5 mg/mL.

Example 13 antibody constant region substitutions

In this example, the heavy chain constant region was replaced with IgG1(N297A/K322A), IgG1(N297A) and IgG2 while maintaining the CR3#30/IgG4 and CR3#75/IgG4 variable regions unchanged, resulting in 6 new antibodies with the following amino acid sequences:

CR3#30/IgG1(N297A/K322A) heavy chain (SEQ ID NO: 25):

QVQLVESGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGTVDDAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK；

CR3#30/IgG1(N297A) heavy chain (SEQ ID NO: 26):

QVQLVESGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGTVDDAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK；

CR3#30/IgG2 heavy chain (SEQ ID NO: 27):

QVQLVESGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGTVDDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG。

the CR3#30/IgG1(N297A/K322A), CR3#30/IgG1(N297A) and CR3#30/IgG2 all adopt CR3#30/lambda light chain, and the sequences are shown as SEQ ID NO: 28:

SEQ ID NO:28：

QSALTQPPSASGSPGQSVTISCTGTSSDVGGYDLVSWYQQHPGKAPKLMIYDVSKRPSGVPDRFSGSKSGNTASLTVSGLQADDEADYYCSSFADSNNWEVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS。

CR3#75/IgG1(N297A/K322A) heavy chain (SEQ ID NO: 29):

EVQLVQSGAEVKKPGATVKISCKVSGYTFTDYYTHWVQQAPGKGLEWMGLVDPEGGETIYAEKFQGRVTITADASTDTAYMELSSLRSEDTAVYYCATGSFLAAPDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK；

CR3#75/IgG1(N297A) heavy chain (SEQ ID NO: 30):

EVQLVQSGAEVKKPGATVKISCKVSGYTFTDYYTHWVQQAPGKGLEWMGLVDPEGGETIYAEKFQGRVTITADASTDTAYMELSSLRSEDTAVYYCATGSFLAAPDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYASTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK；

CR3#75/IgG2 heavy chain (SEQ ID NO: 31):

EVQLVQSGAEVKKPGATVKISCKVSGYTFTDYYTHWVQQAPGKGLEWMGLVDPEGGETIYAEKFQGRVTITADASTDTAYMELSSLRSEDTAVYYCATGSFLAAPDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG。

the CR3#75/IgG1(N297A/K322A), CR3#75/IgG1(N297A) and CR3#75/IgG2 all adopt CR3#75/kappa light chain, and the sequences are shown as SEQ ID NO: 32.

SEQ ID NO:32：

And (3) carrying out gene synthesis on the antibody to construct recombinant plasmids, identifying the recombinant plasmids as positive clones which are correctly inserted into target fragments through sequencing, and preparing sufficient sterile recombinant plasmids by using a plasmid extraction kit for transient transfection to produce protein samples.

Example 14 production of antibodies to different constant regions

A total of 9 recombinant plasmids of CR3#24/IgG4, CR3#30/IgG4, CR3#75/IgG4, CR3#30/IgG1(N297A/K322A), CR3#30/IgG1(N297A), CR3#30/IgG2, CR3#75/IgG1(N297A/K322A), CR3#75/IgG1(N297A) and CR3#75/IgG2 were transfected with a transfection reagent (Invitrogen) to 6X 10⁶Antibody expression was performed in expihho.

Recombinant ExpicHO cells at 37 deg.C, 5% CO₂After medium culture for 9-11 days, the antibody is secreted into serum-free cell culture supernatant. And (3) carrying out ProteinA-HPLC on the fermentation liquor to detect the protein expression quantity. The method comprises the following steps:

samples of the fermentation broth were filtered through a 0.22 μm filter (Sartorius, Cat #17C07FT) and applied to Poros-20A Column (Thermo Scientific, Cat #1-5024-12) for ProteinA-HPLC at 25 ℃ with the chromatographic parameters shown in Table 6. Finally, data collection and analysis were performed using Empower 3, and the results are shown in table 7.

TABLE 6 ProteinA-HPLC chromatogram parameters

TABLE 7 ProteinA-HPLC detection of protein expression level in fermentation broth

After 9 batches of CHO cells are cultured for 9-11 days, SEC-HPLC is carried out on all fermentation liquor to detect the protein purity, and the steps are as follows:

after filtration of the protein sample through a 0.22 μm filter (Sartorius, Cat #17C07FT), the protein sample was applied to a TSKgel G3000 SWXL SEC Column (Tosoh, Cat #008541) and subjected to SEC separation at 25 ℃ using a Waters e2695 detector, a Waters 2998PDA detector as the detector, a 50mM phosphate +400mM sodium chloride (pH 7.0) buffer as the mobile phase, set at a flow rate of 0.5mL/min and a sample volume of 10 μ L, and the absorbance of each fraction was measured at 280 nm. Finally, data acquisition and analysis were performed using Empower 3, and the results are shown in table 8.

TABLE 8 SEC-HPLC DETECTION OF PURITY OF FERMENTATION LIQUID

Sample numbering	Sample name	High molecular weight (%)	Main Peak (%)	Low molecular weight(％)
					S125(38#)-1	CR3#24/IgG4	30.2	19.1	50.7
S125(38#)-2	CR3#30/IgG4	31.6	19.7	48.6
					S125(38#)-3	CR3#75/IgG4	28.4	24.8	46.8
S125(38#)-4	CR3#30/IgG1(N297A/K322A)	29.7	22.7	47.6
					S125(38#)-5	CR3#30/IgG1(N297A)	29.3	25.5	45.2
S125(38#)-6	CR3#30/IgG2	34.6	10.8	54.7
					S125(38#)-7	CR3#75/IgG1(N297A/K322A)	30.0	19.0	51.0
S125(38#)-8	CR3#75/IgG1(N297A)	29.6	19.8	50.5
					S125(38#)-9	CR3#75/IgG2	27.1	13.5	59.5

After the fermentation liquor of 9 candidate molecules is subjected to prism A capture, an ultraviolet spectrophotometer is utilized to measure the protein concentration, and the steps are as follows:

taking the sample solution, and taking the same batch of solvent for preparing the sample solution as a blank control. And (3) diluting the sample by using a blank solvent by using a 1cm quartz absorption cell, and placing the quartz absorption cell filled with the sample into a detection cell for detection. The UV spectrophotometer was IMPLEN, model NP 80.

The protein concentration calculation formula is as follows:

in the formula, X: dilution times;

test proteinThe absorption coefficient of (2).

The results are shown in Table 9.

TABLE 9 concentration of purified samples

Sample numbering	Sample name	Protein concentration (mg/mL)
			S125(38#)-1	CR3#24/IgG4	2.5
S125(38#)-2	CR3#30/IgG4	2.6
			S125(38#)-3	CR3#75/IgG4	2.5
S125(38#)-4	CR3#30/IgG1(N297A/K322A)	4.4
			S125(38#)-5	CR3#30/IgG1(N297A)	2.4
S125(38#)-6	CR3#30/IgG2	1.4
			S125(38#)-7	CR3#75/IgG1(N297A/K322A)	3.0
S125(38#)-8	CR3#75/IgG1(N297A)	2.0
			S125(38#)-9	CR3#75/IgG2	1.4

The fermentation liquor of 9 candidate molecules is captured by prism A and then analyzed by SEC-HPLC, and the steps are as follows:

after filtration of the protein sample through a 0.22 μm filter (Sartorius, Cat #17C07FT), the protein sample was applied to a TSKgel G3000 SWXL SEC Column (Tosoh, Cat #008541) and subjected to SEC separation at 25 ℃ using a Waters e2695, Waters 2998PDA detector as the detector, 50mM phosphate +400mM sodium chloride (pH 7.0) buffer as the mobile phase, set at a flow rate of 0.5mL/min and a sample volume of 10 μ L, and the absorbance was measured at 280 nm. Finally, data acquisition and analysis were performed using Empower 3, and the results are shown in table 10.

TABLE 10 SEC-HPLC DETECTION OF PURIFIED SAMPLE PURITY

By integrating the results, the 9 antibodies can smoothly complete sample production, the concentration is more than 1mg/mL, the purity is higher than 90%, and subsequent experiments can be carried out.

Example 15 aggregative Property analysis

In this example, the isoelectric point of each candidate molecule was determined using iCIEF as follows:

after desalting the sample, it was diluted with distilled water to a protein concentration of about 2mg/mL and prepared as follows (200. mu.L system): 8 μ L of 2% pharmalyte (pH 3-10) (GE, Cat #17045601), 70 μ L of 1% MC (ProteinSimple, Cat #1018761), 1 μ L of Low pI (ProteinSimple, Cat #102222), 1 μ L of High pI (ProteinSimple, Cat #101996), 100 μ L of 8M urea (Sigma, Cat # U0631-500G1), 20 μ L of sample.

The system is arranged in a centrifuge tube, then centrifuged for 5min at 10000g, supernatant (60-120 mu L) is taken and added into an inner cannula, the inner cannula is placed in the centrifuge tube, centrifuged for 5min at 10000g, the inner cannula is taken out, placed in a sample bottle and tested on a computer, and the equipment is protein simple iCE 3. As a result, as shown in Table 11, the actual isoelectric point was slightly higher than the predicted isoelectric point.

TABLE 11 isoelectric points of candidate molecules

Sample numbering	Sample name	Prediction of isoelectric point	iCIEF-isoelectric point
				S125(38#)-1	CR3#24/IgG4	5.97	6.13
S125(38#)-2	CR3#30/IgG4	6.18	6.30
				S125(38#)-3	CR3#75/IgG4	5.92	5.89
S125(38#)-4	CR3#30/IgG1(N297A/K322A)	7.24	7.33
				S125(38#)-5	CR3#30/IgG1(N297A)	7.56	7.57
S125(38#)-6	CR3#30/IgG2	6.50	6.97
				S125(38#)-7	CR3#75/IgG1(N297A/K322A)	6.36	6.58
S125(38#)-8	CR3#75/IgG1(N297A)	6.51	6.76
				S125(38#)-9	CR3#75/IgG2	5.97	6.18

EXAMPLE 16 capillary electrophoresis analysis

After prism A capture of the fermentation broth of 9 candidate molecules, CE-SDS analysis was performed, the procedure was as follows:

protein samples were ultrafiltered in 30 μm ultrafiltration centrifuge tubes (Sartorius, Cat # VS0122) and samples were prepared, heated at 70 ℃ and loaded, and CE-SDS separation was performed at 25 ℃ using SCIEX PA800 Plus as the apparatus and PDA as the detector, and absorbance was measured at 220 nm. Data collection and analysis were finally performed using 32karat, and the results are shown in Table 12, where CR3#75/IgG2 was less pure.

TABLE 12 capillary electrophoresis for purity determination of purified samples

Example 17 protein stability assay for C5-specific antibodies

This example employs a protein stability analysis system

Tm of (Uncariamed Labs, Calif.)&Tagg with optional DLS program, protein particle size and stability. Protein particle size and particle size distribution and thermal stability were analyzed from multiple angles by analyzing the resulting parameters Tm (protein melting temperature), Tagg266 (protein aggregation temperature), Tagg473 (protein aggregation temperature), Z-Ave. Dia (protein average particle size), PDI (polydispersity index), Pk1 Mode Dia, Pk 1Mass (%), Pk2 Mode Dia and Pk2 Mass (%) 9 parameters. The method comprises the following steps:

mu.L of antibody was added to uni tubes (Uncariamed Labs, Calif.) and 2 replicates of each antibody were run using the Tm & Tagg with optional DLS program.

The particle size results at 25 ℃ and 95 ℃ are shown in tables 13 and 14, respectively, and the Tm & Tagg results are shown in Table 15. Wherein, Z-Ave.Dia-sample average hydration kinetic diameter reflects the overall particle size condition of the sample; PDI-polydispersity index, PDI <0.1 indicates narrow particle size distribution, with larger values indicating more non-uniform particle size distribution and higher degree of dispersion; fit Var-fitting deviation, less than 0.01 indicates high credibility of Z-Ave.Dia and PDI, and the larger the value is, the lower the credibility is; pk1 Mode Dia-light intensity distribution shows multi-peak, the Mode particle size of the first main peak; pk 1Mass (%) -Mass ratio of the first peak.

As can be seen from Table 13, the average kinetic diameters of hydration of S125(38#) -3/6/7/9 and Pk1 model Dia. are both around 11nm, no large particle size component is present, PDI <0.1 indicates uniform particle size, and protein monomers are presumed; the remaining samples in the group all had larger particle size components, presumably aggregates. As can be seen from Table 14, the protein molecules produced significant aggregation upon heating, as seen by the average hydration kinetic diameters and PDI. As can be seen from Table 15, S125 # (38#) -4 and S125 #) -7Tm are high, and the conformational thermal stability is expected to be high, and S125 # (38#) -2/4/5/6/7Tagg266 is at 60 ℃ or higher, and the colloidal thermal stability is expected to be high.

TABLE 1325 ℃ particle size results

TABLE 1495 ℃ particle size results

TABLE 15 Tm & Tagg results

From the above results, it can be seen that the overall thermal stability of the S125(38#) -7 group is high, that is, CR3#75/IgG1(N297A/K322A) has thermal stability superior to CR3#75/IgG 4.

In conclusion, the anti-C5 antibody has remarkable binding capacity with the humanized recombinant C5 protein, long half-life period, good solubility and good thermal stability, is expected to be developed into a protein drug and is applied to preventing and/or treating autoimmune diseases caused by over-activation of C5.

The applicant states that the present invention is illustrated in detail by the above examples, but the present invention is not limited to the above detailed methods, i.e. it is not meant that the present invention must rely on the above detailed methods for its implementation. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.

Sequence listing

<110> Tianshili biomedical products Ltd

<120> anti-C5 antibody and use thereof

<130> 2021

<160> 32

<170> PatentIn version 3.3

<210> 1

<211> 5

<212> PRT

<213> Artificial sequence

<400> 1

Asp Tyr Tyr Met His

1 5

<210> 2

<211> 17

<212> PRT

<213> Artificial sequence

<400> 2

Val Val Asp Pro Glu Asp Gly Glu Ile Ile Tyr Ala Glu Lys Phe Gln

1 5 10 15

Gly

<210> 3

<211> 12

<212> PRT

<213> Artificial sequence

<400> 3

Ser Asp Tyr Gly Ala Gly Ser Tyr Met Val Asp Pro

1 5 10

<210> 4

<211> 11

<212> PRT

<213> Artificial sequence

<400> 4

Gly Gly Asn Asn Leu Glu Ser Lys Ile Val Ser

1 5 10

<210> 5

<211> 7

<212> PRT

<213> Artificial sequence

<400> 5

Tyr Asp Thr Asp Arg Pro Ser

1 5

<210> 6

<211> 11

<212> PRT

<213> Artificial sequence

<400> 6

Gln Val Trp Asp Ser Asn Ser Asp His Val Ile

1 5 10

<210> 7

<211> 5

<212> PRT

<213> Artificial sequence

<400> 7

Ser Tyr Tyr Met His

1 5

<210> 8

<211> 17

<212> PRT

<213> Artificial sequence

<400> 8

Ile Ile Asn Pro Ser Gly Gly Ser Thr Ser Tyr Ala Gln Lys Phe Gln

1 5 10 15

Gly

<210> 9

<211> 9

<212> PRT

<213> Artificial sequence

<400> 9

Gly Thr Val Asp Asp Ala Phe Asp Ile

1 5

<210> 10

<211> 14

<212> PRT

<213> Artificial sequence

<400> 10

Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr Asp Leu Val Ser

1 5 10

<210> 11

<211> 7

<212> PRT

<213> Artificial sequence

<400> 11

Asp Val Ser Lys Arg Pro Ser

1 5

<210> 12

<211> 12

<212> PRT

<213> Artificial sequence

<400> 12

Ser Ser Phe Ala Asp Ser Asn Asn Trp Glu Val Val

1 5 10

<210> 13

<211> 5

<212> PRT

<213> Artificial sequence

<400> 13

Asp Tyr Tyr Thr His

1 5

<210> 14

<211> 17

<212> PRT

<213> Artificial sequence

<400> 14

Leu Val Asp Pro Glu Gly Gly Glu Thr Ile Tyr Ala Glu Lys Phe Gln

1 5 10 15

Gly

<210> 15

<211> 9

<212> PRT

<213> Artificial sequence

<400> 15

Gly Ser Phe Leu Ala Ala Pro Asp Tyr

1 5

<210> 16

<211> 17

<212> PRT

<213> Artificial sequence

<400> 16

Lys Ser Ser Gln Ser Leu Leu Tyr Asn Ser Asn Asn Lys Asn Tyr Leu

1 5 10 15

Ala

<210> 17

<211> 7

<212> PRT

<213> Artificial sequence

<400> 17

Trp Ala Ser Thr Arg Glu Ser

1 5

<210> 18

<211> 9

<212> PRT

<213> Artificial sequence

<400> 18

Gln Gln Tyr Tyr Ser Thr Pro Leu Thr

1 5

<210> 19

<211> 448

<212> PRT

<213> Artificial sequence

<400> 19

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ala Val Lys Ile Ser Cys Lys Val Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Tyr Met His Trp Val Gln Gln Ala Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Val Val Asp Pro Glu Asp Gly Glu Ile Ile Tyr Ala Glu Lys Phe

50 55 60

Gln Gly Arg Ile Thr Ile Thr Ala Asp Thr Ser Thr Asp Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Ser Asp Tyr Gly Ala Gly Ser Tyr Met Val Asp Pro Trp Gly

100 105 110

Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser

115 120 125

Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala

130 135 140

Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val

145 150 155 160

Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala

165 170 175

Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val

180 185 190

Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His

195 200 205

Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly

210 215 220

Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro

260 265 270

Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440 445

<210> 20

<211> 214

<212> PRT

<213> Artificial sequence

<400> 20

Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Thr Pro Gly Lys

1 5 10 15

Thr Ala Arg Ile Ala Cys Gly Gly Asn Asn Leu Glu Ser Lys Ile Val

20 25 30

Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Phe

35 40 45

Tyr Asp Thr Asp Arg Pro Ser Gly Ile Ser Glu Arg Phe Ser Gly Ser

50 55 60

Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Arg Val Glu Ala Gly

65 70 75 80

Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Asn Ser Asp His

85 90 95

Val Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro Lys

100 105 110

Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu Gln

115 120 125

Ala Asn Lys Ala Thr Leu Val Cys Leu Val Ser Asp Phe Tyr Pro Gly

130 135 140

Ala Val Thr Val Ala Trp Lys Ala Asp Gly Ser Pro Val Lys Val Gly

145 150 155 160

Val Glu Thr Thr Lys Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala Ala

165 170 175

Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg Ser

180 185 190

Tyr Ser Cys Arg Val Thr His Glu Gly Ser Thr Val Glu Lys Thr Val

195 200 205

Ala Thr Ala Glu Cys Ser

210

<210> 21

<211> 445

<212> PRT

<213> Artificial sequence

<400> 21

Gln Val Gln Leu Val Glu Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Ile Ile Asn Pro Ser Gly Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Thr Val Asp Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys

210 215 220

Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu

225 230 235 240

Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu

245 250 255

Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln

260 265 270

Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys

275 280 285

Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu

290 295 300

Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys

305 310 315 320

Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys

325 330 335

Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser

340 345 350

Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys

355 360 365

Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln

370 375 380

Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly

385 390 395 400

Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln

405 410 415

Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn

420 425 430

His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440 445

<210> 22

<211> 218

<212> PRT

<213> Artificial sequence

<400> 22

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asp Leu Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Asp Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ala Asp Ser

85 90 95

Asn Asn Trp Glu Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser

115 120 125

Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp

130 135 140

Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro

145 150 155 160

Val Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn

165 170 175

Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Glu

180 185 190

Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val

195 200 205

Glu Lys Thr Val Ala Pro Thr Glu Cys Ser

210 215

<210> 23

<211> 445

<212> PRT

<213> Artificial sequence

<400> 23

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Val Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Tyr Thr His Trp Val Gln Gln Ala Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Leu Val Asp Pro Glu Gly Gly Glu Thr Ile Tyr Ala Glu Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Ala Ser Thr Asp Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Thr Gly Ser Phe Leu Ala Ala Pro Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro Cys

210 215 220

Pro Pro Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu

225 230 235 240

Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu

245 250 255

Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln

260 265 270

Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys

275 280 285

Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu

290 295 300

Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys

305 310 315 320

Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys

325 330 335

Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser

340 345 350

Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys

355 360 365

Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln

370 375 380

Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly

385 390 395 400

Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln

405 410 415

Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn

420 425 430

His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

435 440 445

<210> 24

<211> 220

<212> PRT

<213> Artificial sequence

<400> 24

Asp Ile Val Met Thr Gln Ala Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Val Asn Cys Lys Ser Ser Gln Ser Leu Leu Tyr Asn

20 25 30

Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Phe Cys Gln Gln

85 90 95

Tyr Tyr Ser Thr Pro Leu Thr Leu Gly Gly Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

<210> 25

<211> 448

<212> PRT

<213> Artificial sequence

<400> 25

Gln Val Gln Leu Val Glu Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Ile Ile Asn Pro Ser Gly Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Thr Val Asp Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Ala Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 26

<211> 448

<212> PRT

<213> Artificial sequence

<400> 26

Gln Val Gln Leu Val Glu Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Ile Ile Asn Pro Ser Gly Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Thr Val Asp Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 27

<211> 443

<212> PRT

<213> Artificial sequence

<400> 27

Gln Val Gln Leu Val Glu Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr

20 25 30

Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Ile Ile Asn Pro Ser Gly Gly Ser Thr Ser Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Thr Val Asp Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr

100 105 110

Met Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys

210 215 220

Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe

225 230 235 240

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

245 250 255

Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe

260 265 270

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

275 280 285

Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr

290 295 300

Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

305 310 315 320

Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr

325 330 335

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg

340 345 350

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

355 360 365

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

370 375 380

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser

385 390 395 400

Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln

405 410 415

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

420 425 430

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440

<210> 28

<211> 218

<212> PRT

<213> Artificial sequence

<400> 28

Gln Ser Ala Leu Thr Gln Pro Pro Ser Ala Ser Gly Ser Pro Gly Gln

1 5 10 15

Ser Val Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr

20 25 30

Asp Leu Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu

35 40 45

Met Ile Tyr Asp Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe

50 55 60

Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu

65 70 75 80

Gln Ala Asp Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ala Asp Ser

85 90 95

Asn Asn Trp Glu Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu

100 105 110

Gly Gln Pro Lys Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser

115 120 125

Glu Glu Leu Gln Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp

130 135 140

Phe Tyr Pro Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro

145 150 155 160

Val Lys Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn

165 170 175

Lys Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys

180 185 190

Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val

195 200 205

Glu Lys Thr Val Ala Pro Thr Glu Cys Ser

210 215

<210> 29

<211> 448

<212> PRT

<213> Artificial sequence

<400> 29

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Val Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Tyr Thr His Trp Val Gln Gln Ala Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Leu Val Asp Pro Glu Gly Gly Glu Thr Ile Tyr Ala Glu Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Ala Ser Thr Asp Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Thr Gly Ser Phe Leu Ala Ala Pro Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Ala Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 30

<211> 448

<212> PRT

<213> Artificial sequence

<400> 30

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Val Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Tyr Thr His Trp Val Gln Gln Ala Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Leu Val Asp Pro Glu Gly Gly Glu Thr Ile Tyr Ala Glu Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Ala Ser Thr Asp Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Thr Gly Ser Phe Leu Ala Ala Pro Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr

210 215 220

His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser

225 230 235 240

Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg

245 250 255

Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro

260 265 270

Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala

275 280 285

Lys Thr Lys Pro Arg Glu Glu Gln Tyr Ala Ser Thr Tyr Arg Val Val

290 295 300

Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr

305 310 315 320

Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr

325 330 335

Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu

340 345 350

Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys

355 360 365

Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser

370 375 380

Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp

385 390 395 400

Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser

405 410 415

Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala

420 425 430

Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys

435 440 445

<210> 31

<211> 443

<212> PRT

<213> Artificial sequence

<400> 31

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Thr Val Lys Ile Ser Cys Lys Val Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Tyr Thr His Trp Val Gln Gln Ala Pro Gly Lys Gly Leu Glu Trp Met

35 40 45

Gly Leu Val Asp Pro Glu Gly Gly Glu Thr Ile Tyr Ala Glu Lys Phe

50 55 60

Gln Gly Arg Val Thr Ile Thr Ala Asp Ala Ser Thr Asp Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Thr Gly Ser Phe Leu Ala Ala Pro Asp Tyr Trp Gly Gln Gly Thr

100 105 110

Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro

115 120 125

Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly

130 135 140

Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn

145 150 155 160

Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln

165 170 175

Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser

180 185 190

Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser

195 200 205

Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys

210 215 220

Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe

225 230 235 240

Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val

245 250 255

Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe

260 265 270

Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro

275 280 285

Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr

290 295 300

Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val

305 310 315 320

Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr

325 330 335

Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg

340 345 350

Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly

355 360 365

Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro

370 375 380

Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser

385 390 395 400

Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln

405 410 415

Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His

420 425 430

Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440

<210> 32

<211> 220

<212> PRT

<213> Artificial sequence

<400> 32

Asp Ile Val Met Thr Gln Ala Pro Asp Ser Leu Ala Val Ser Leu Gly

1 5 10 15

Glu Arg Ala Thr Val Asn Cys Lys Ser Ser Gln Ser Leu Leu Tyr Asn

20 25 30

Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln

35 40 45

Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val

50 55 60

Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr

65 70 75 80

Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Phe Cys Gln Gln

85 90 95

Tyr Tyr Ser Thr Pro Leu Thr Leu Gly Gly Gly Thr Lys Val Glu Ile

100 105 110

Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp

115 120 125

Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn

130 135 140

Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu

145 150 155 160

Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp

165 170 175

Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr

180 185 190

Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser

195 200 205

Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys

210 215 220

Claims

1. An anti-C5 antibody, wherein said anti-C5 antibody comprises a heavy chain variable region and a light chain variable region;

2. The anti-C5 antibody of claim 1, wherein the heavy chain variable region further comprises CDR1 shown in SEQ ID NO 1, SEQ ID NO 7 or SEQ ID NO 13;

preferably, the heavy chain variable region further comprises CDR2 shown in SEQ ID NO 2, SEQ ID NO 8 or SEQ ID NO 14;

preferably, the light chain variable region further comprises CDR1 shown in SEQ ID NO. 4, SEQ ID NO. 10 or SEQ ID NO. 16;

3. The anti-C5 antibody according to claim 1 or 2, wherein the heavy chain variable region of the anti-C5 antibody comprises CDR1 shown in SEQ ID NO:1, CDR2 shown in SEQ ID NO:2, CDR3 shown in SEQ ID NO: 3;

the light chain variable region of the anti-C5 antibody comprises CDR1 shown in SEQ ID NO. 4, CDR2 shown in SEQ ID NO. 5, and CDR3 shown in SEQ ID NO. 6;

preferably, the heavy chain variable region of the anti-C5 antibody includes CDR1 shown in SEQ ID NO. 7, CDR2 shown in SEQ ID NO. 8, CDR3 shown in SEQ ID NO. 9;

the light chain variable region of the anti-C5 antibody includes CDR1 shown in SEQ ID NO. 10, CDR2 shown in SEQ ID NO. 11, CDR3 shown in SEQ ID NO. 12;

preferably, the heavy chain variable region of the anti-C5 antibody includes CDR1 shown in SEQ ID NO. 13, CDR2 shown in SEQ ID NO. 14, CDR3 shown in SEQ ID NO. 15;

the variable region of the light chain of the anti-C5 antibody includes CDR1 shown in SEQ ID NO. 16, CDR2 shown in SEQ ID NO. 17, and CDR3 shown in SEQ ID NO. 18.

4. The anti-C5 antibody of any one of claims 1-3, wherein the heavy chain variable region of the anti-C5 antibody comprises the amino acid sequence set forth in SEQ ID NO 19 and the light chain variable region comprises the amino acid sequence set forth in SEQ ID NO 20;

preferably, the heavy chain variable region of the anti-C5 antibody comprises the amino acid sequence shown in SEQ ID NO. 21, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO. 22;

preferably, the heavy chain variable region of the anti-C5 antibody comprises the amino acid sequence shown in SEQ ID NO. 23, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO. 24;

preferably, the anti-C5 antibody further comprises a constant region;

preferably, the constant region is derived from any one of IgG4, IgG1, or IgG 2;

preferably, the IgG4 has the S228P mutation;

preferably, the IgG1 has the N297A and/or K322A mutation;

preferably, the IgG2 has the S257A mutation.

5. A nucleic acid molecule comprising a DNA fragment encoding the anti-C5 antibody of any one of claims 1-4.

6. An expression vector comprising the nucleic acid molecule of claim 5.

7. A recombinant cell expressing the anti-C5 antibody of any one of claims 1-4;

preferably, the recombinant cell has integrated into its genome the nucleic acid molecule of claim 5;

preferably, the recombinant cell comprises the expression vector of claim 6.

8. A method of producing the anti-C5 antibody of any one of claims 1-4, comprising the steps of:

9. A pharmaceutical composition comprising the anti-C5 antibody of any one of claims 1-4;

preferably, the pharmaceutical composition further comprises an anti-tumor drug;

10. Use of the anti-C5 antibody of any one of claims 1-4, the nucleic acid molecule of claim 5, the expression vector of claim 6, the recombinant cell of claim 7, or the pharmaceutical composition of claim 9 in the preparation of a disease detection reagent and/or a disease treatment drug;