Monoclonal antibody targeting human complement C5 and application thereof

Abstract

The invention relates to a monoclonal antibody targeting human complement C5 and application thereof, the monoclonal antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3. The antibody or the antigen binding fragment thereof has very strong binding capacity with the human complement C5, and blocks the cleavage activation effect of C5 invertase on the complement C5. In addition, C5 activation mediated by complement classical pathway activation and alternative pathway activation can be blocked at the same time.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine and relates to a monoclonal antibody targeting human complement C5 and an application thereof. Background Art

[0002] As an important component of innate immunity, the complement system participates in host defense, inflammation regulation, and tissue homeostasis maintenance through cascade reactions. Among them, complement component C5 is a key node in the terminal complement pathway, and its cleavage products C5a (a potent chemokine) and C5b (a precursor of the membrane attack complex C5b-9) play a core role in various pathological processes. Abnormally activated C5 can drive the progression of various immune-related diseases by recruiting inflammatory cells, mediating endothelial damage, and directly lysing target cells. Studies have shown that excessive activation of the C5 signaling pathway is closely related to diseases such as paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS), myasthenia gravis (gMG), and neuromyelitis optica spectrum disorder (NMOSD), and is significantly associated with pathological processes such as chronic inflammatory diseases, ischemia-reperfusion injury, and age-related macular degeneration (AMD).

[0003] Currently, targeted therapies for C5 (such as anti-C5 monoclonal antibodies Eculizumab, Ravulizumab and Crovalimab, etc.) have shown significant clinical efficacy by blocking C5 cleavage, but their application still faces many challenges: including decreased patient compliance due to frequent dosing, heterogeneity of drug responses due to genetic polymorphisms in some patients, and the risk of infection that may be caused by long-term inhibition of C5. Therefore, the development of new C5 inhibitors with improved pharmacokinetic properties, higher target affinity or the ability to overcome individual differences has become an important research direction for improving clinical efficacy and expanding the scope of indications. The C5 monoclonal antibody inhibitor of the present invention aims to provide a better therapeutic window and disease indication control potential through innovative epitope selection and antibody engineering technology. Summary of the Invention

[0004] In view of the shortcomings of the existing technology, the purpose of the present invention is to provide a monoclonal antibody targeting human complement C5 and its application.

[0005] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides a monoclonal antibody targeting human complement C5, wherein the monoclonal antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3.

[0007] The amino acid sequence of the HCDR1 includes the sequence shown in SEQ ID NO.1, the amino acid sequence of the HCDR2 includes the sequence shown in SEQ ID NO.2, and the amino acid sequence of the HCDR3 includes the sequence shown in SEQ ID NO.3;

[0008] The amino acid sequence of the LCDR1 includes the sequence shown in SEQ ID NO.4, the amino acid sequence of the LCDR2 includes the sequence shown in SEQ ID NO.5, and the amino acid sequence of the LCDR3 includes the sequence shown in SEQ ID NO.6.

[0009] Optionally, the amino acid sequence of the heavy chain variable region includes the sequence shown in SEQ ID NO.7.

[0010] Optionally, the amino acid sequence of the light chain variable region includes the sequence shown in SEQ ID NO.8.

[0011] In a second aspect, the present invention provides a nucleic acid molecule encoding the monoclonal antibody targeting human C5 according to the first aspect.

[0012] Preferably, the nucleotide sequence of the nucleic acid molecule includes the sequence shown in SEQ ID NO.9 and the sequence shown in SEQ ID NO.10.

[0013] In a third aspect, the present invention provides an expression vector comprising the nucleic acid molecule described in the second aspect.

[0014] In a fourth aspect, the present invention provides a host cell, wherein the host cell contains the expression vector described in the third aspect.

[0015] In a fifth aspect, the present invention provides a use of the monoclonal antibody targeting human complement C5 according to the first aspect or the expression vector according to the third aspect in preparing a detection reagent, wherein the target of the detection reagent is human complement C5.

[0016] In a sixth aspect, the present invention provides a pharmaceutical composition comprising the monoclonal antibody targeting human complement C5 as described in the first aspect, and a pharmaceutically acceptable carrier and / or diluent.

[0017] In the seventh aspect, the present invention provides an application of any one or a combination of at least two of the monoclonal antibody targeting human complement C5 according to the first aspect, the expression vector according to the third aspect, the host cell according to the fourth aspect, or the pharmaceutical composition according to the sixth aspect in the preparation of drugs for preventing, improving, and treating autoimmune diseases or drugs for inhibiting immune rejection.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] This antibody or its antigen-binding fragment has strong binding affinity to human complement C5 and blocks the enzymatic cleavage of C5 by complement C5 convertase. It also inhibits the classical and alternative complement signal transduction pathways and their downstream regulatory signals, blocking the activation of C5 by upstream activation signals. This antibody lacks ADCC, ADCP, and CDC effects, preventing immune cells from killing and phagocytosing target cells containing C5 after binding to C5. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is the test result of the binding activity of humanized antibody 6E3-H1L1 to complement C5;

[0021] Figure 2 It is the ability of the humanized antibody 6E3-H1L1 to block activation of the classical complement pathway;

[0022] Figure 3 It is the ability of the humanized antibody 6E3-H1L1 to block activation of the alternative classical pathway;

[0023] Figure 4 It is the humanized antibody 6E3-H1L1 that mediates ADCC. DETAILED DESCRIPTION

[0024] The technical solution of the present invention is further described below by way of specific embodiments. It should be understood by those skilled in the art that the embodiments are merely to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0025] Example 1

[0026] Preparation of mouse antibody against human complement C5

[0027] Eight-week-old female Balb / c mice were immunized with the full-length human complement C5 (P01031) protein sequence as the immunizing antigen. For the first immunization, purified antigen was thoroughly emulsified with complete Freund's adjuvant and injected subcutaneously at six injection sites, with a volume of 50 μL per site. For the second, third, and fourth immunizations, antigen was thoroughly emulsified with incomplete adjuvant and injected subcutaneously at six injection sites, with a volume of 50 μL per site. Seven days after the fourth immunization, blood was collected from the tail vein of the mice for titer determination. Mice with the highest titer were selected for a booster immunization via intraperitoneal injection of 25 μg of antigen per mouse. Three days later, spleens were harvested under sterile conditions. Spleens were dissected, minced, and prepared into single-cell suspensions. Splenocytes were mixed at a ratio of 10:1, and the harvested splenocytes were washed twice with IMEM medium. After centrifugation of the mixed cells, the supernatant was removed and PEG1500 was slowly added to the cell pellet. The pellet was gently shaken and incubated for 2 minutes. The cell fusion reaction was terminated by adding 2 mL of IMEM. Then, 10 mL of IMEM was slowly added and the pellet was centrifuged at 800 rpm, and the supernatant was discarded. The cells were then thoroughly resuspended in IMEM (1×HAT) containing the double-antibody and 15% FBS. 300 μL of the cell suspension was added to each well of a 96-well cell culture plate. The 96-well plate was incubated in a 37°C cell culture incubator. Hybridoma clones were obtained after 10 days. Positive clones were isolated by limiting dilution to obtain single-cell positive clones. These clones were expanded and the cell culture supernatant was harvested. The cell supernatant was purified using a Protein A / G affinity chromatography column to obtain anti-human complement C5 protein antibodies, resulting in the human complement C5 mouse antibody m6E3.

[0028] The amino acid sequence of the heavy chain variable region of m6E3 is shown in SEQ ID NO.11.

[0029] SEQ ID NO.11:

[0030] EVQLQQSGPELVKPGASVKISCKTS GYTFTENT MLWVKQSHGKSLEWIG G IIPNNGDT DYNQKFKGKATLTVDRSSSTAYMELRRSLTSEDSAVYYC ARSLLR LLPFDY WGQGTTLTVSS.

[0031] The amino acid sequence of HCDR1 in the heavy chain variable region is shown in SEQ ID NO.1, the amino acid sequence of HCDR2 is shown in SEQ ID NO.2, and the amino acid sequence of HCDR3 is shown in SEQ ID NO.3.

[0032] SEQ ID NO.1: GYTFTENT .

[0033] SEQ ID NO.2: IIPNNGDT .

[0034] SEQ ID NO.3: ARSLLRLLPFDY .

[0035] Optionally, the amino acid sequence of the heavy chain variable region includes the sequence shown in SEQ ID NO.7:

[0036] QMQLVQSGAEVKKPGSSVKVSCKAS GYSFISFY MHWVRQAHGQALEWI GY IDPFNGDT NYAQKFQDRVTITVDKSSTTAYMELSSLTSEDSSVYYC TRSGA YGHYGY WGQGTTVTVSS.

[0037] Optionally, the amino acid sequence of the light chain variable region includes the sequence shown in SEQ ID NO.8.

[0038] SEQ ID NO.8:

[0039] DIVLTQSPASLAVSPGQRATITCRAS ESVDSYDNSF MHWYQQKPGQPPKL LIY LVS NLDSGVPARFSGSGSGTDFTLTINPVEADDTATYYC HQNKEYPLT FGG GTKLELK.

[0040] The present invention provides an expression vector. The nucleotide sequence of the nucleic acid molecule contained in the expression vector includes the sequence shown in SEQ ID NO.9 and the sequence shown in SEQ ID NO.10.

[0041] SEQ ID NO.9:

[0042] CAGATGCAGCTGGTGCAGAGCGGCGCGGAAGTGAAAAAACCGGGCAGCAGCGTGAAAGTGAGCTGCAAAGCGAGCGGCTATAGCTTTATTAGCTTTTATATGCATTGGGTGCGCCAGGCGCATGGCCAGGCGCTGGAATGGATTGGCTATATTGATCCGTTTAACGGCGATACCAACTATGCGCAGAAATTTCAGGATCGCGTGACCATTACCGTGGATAAAAGCAGCACCACCGCGTATATGGAACTGAGCAGCCTGACCAGCGAAGATAGCAGCGTGTATTATTGCACCCGCAGCGGCGCGTATGGCCATTATGGCTATTGGGGCCAGGGCACCACCGTGACCGTGAGCAGC。

[0043] SEQ ID NO.10:

[0044] GATATTGTGCTGACCCAGAGCCCGGCGAGCCTGGCGGTGAGCCCGGGCCAGCGCGCGACCATTACCTGCCGCGCGAGCGAAAGCGTGGATAGCTATGATAACAGCTTTATGCATTGGTATCAGCAGAAACCGGGCCAGCCGCCGAAACTGCTGATTTATCTGGTGAGCAACCTGGATAGCGGCGTGCCGGCGCGCTTTAGCGGCAGCGGCAGCGGCACCGATTTTACCCTGACCATTAACCCGGTGGAAGCGGATGATACCGCGACCTATTATTGCCATCAGAACAAAGAATATCCGCTGACCTTTGGCGGCGGCACCAAACTGGAACTGAAA。

[0045] The nucleotide sequence of the heavy chain variable region of the indicated m6E3 is shown in SEQ ID NO.12.

[0046] SEQ ID NO.12:

[0047] GATATTGTGATGACCCAGAGCCATAAATTTATGAGCACCAGCGTGGGGCGATCGCGTGAGCATTACCTGCAAAGCGAGCCAGGATGTGAGCACCGCGGTGGCGTGGTATCAGCAGAAACCGGGCCTGAGCCCGAAACTGCTGATTTATTGGGCGAGCACCCGC CATACCGGCGTGCCGGATCGCTTTACCGGCAGCGGCAGCGGCACCGATTTTACCCTGACCATTAGCAGCGTGCAGGCGGAAGATCTGGAACTGTATTATTGCCAGCAGCATTATGATACCCCGTGGACCTTTGGCGGCGGCACCAAACTGGATATTAAACGC.

[0048] The amino acid sequence of the light chain variable region of M6E3 is shown in SEQ ID NO.13.

[0049] SEQ ID NO.13:

[0050] DIVMTQSHKFMSTSVGDRVSITCKAS QDVSTA VAWYQQKPGLSPKLLIY WAS TRHTGVPDRFTGSGSGTDFTLTISSVQAEDLELYYC QQHYDTPWT FGGG TKLDIKR.

[0051] The amino acid sequence of LCDR1 in the light chain variable region is shown in SEQ ID NO.4, the amino acid sequence of LCDR2 is shown in SEQ ID NO.5, and the amino acid sequence of LCDR3 is shown in SEQ ID NO.6.

[0052] SEQ ID NO.4: QDVSTA .

[0053] SEQ ID NO.5: WAS .

[0054] SEQ ID NO.6: QQHYDTPW .

[0055] The nucleotide sequence of the light chain variable region of m6E3 is shown in SEQ ID NO.14.

[0056] SEQ ID NO.14:

[0057] GAAGTGCAGCTGCAGCAGAGCGGCCCGGAACTGGTGAAACCGGGCGCGAGCGTGAAAATTAGCTGCAAAACCAGCGGCTATAACCTTTACCGAAAACACCATGCTGTGGGTGAAACAGAGCCATGGCAAAAGCCTGGAATGGATTGGCGGCATTATTCCGAACAACGGCGATACCGATTA TAACCAGAAATTTAAAGGCAAAGCGACCCTGACCGTGGATCGCAGCAGCAGCACCGCGTATATGGAACTGCGCAGCCTGACCAGCGAAGATAGCGCGGTGTATTATTGCGCGCGCAGCCTGCTGCGCCTGCTGCCGTTTGATTGGGGCCAGGGCACCACCCTGACCGTGAGCAGC.

[0058] Example 2

[0059] Preparation of humanized antibody against human complement C5

[0060] For the heavy and light chain variable region sequences of the m6E3 antibody, human antibody frameworks closest to the murine framework sequences were selected for CDR grafting. Humanizing mutations were performed on the amino acid sequences of portions of the FW region, and backmutations were performed on residues critical for VH and VL conformation, resulting in the humanized antibodies 6E3-HwLw, 6E3-H1L1, and 6E3-H2L1. The heavy chain sequence of the humanized antibody 6E3-H1L1 is shown in SEQ ID NO. 15, and the amino acid sequence of the light chain variable region includes the sequence shown in SEQ ID NO. 16.

[0061] SEQ ID NO.15:

[0062] QVQLVQSGAEVKKPGASVKVSCKTSGYTFTENTMLWVKQAPGQGLEWIGGIIPNNGDTDYNQKFKGRVTLTRDTSSSTAYMELSRLTSEDTAVYYCARSLLRLLPFDYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK。

[0063] SEQ ID NO.16:

[0064] DIVMTQSPDSLAVSLGERATINCKASQDVSTAVAWYQQKPGQPPKLLIYWASTRHTGVPDRFSGSGSGTDFTLTISSLQAEDVEVYYCQQHYDTPWTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC。

[0065] The nucleotide sequences of the heavy and light chains of the indicated 6E3-H1L1 are shown in SEQ ID NO.17 and SEQ ID NO.18.

[0066] SEQ ID NO.17

[0067]

[0068] SEQ ID NO.18

[0069] GATATTGTGATGACCCAGAGCCCGGATAGCCTGGCGGTGAGCCTGGGCGAACGCGCGACCATTAACTGCAAAGCGAGCCAGGATGTGAGCACCGCGGTGGCGTGGTATCAGCAGAAACCGGGCCAGCCGCCGAAACTGCTGATTTATTGGGCGAGCACCCGCCATACCGGCGTGCCGGATCGCTTTAGCGGCAGCGGCAGCGGCACCGATTTTACCCTGACCATTAGCAGCCTGCAGGCGGAAGATGTGGAAGTGTATTATTGCCAGCAGCATTATGATACCCCGTGGACCTTTGGCGGCGGCACCAAACTGGAAATTAAACGCACCGTGGCGGCGCCGAGCGTGTTTATTTTTCCGCCGAGCGATGAACAGCTGAAAAGCGGCACCGCGAGCGTGGTGTGCCTGCTGAACAACTTTTATCCGCGCGAAGCGAAAGTGCAGTGGAAAGTGGATAACGCGCTGCAGAGCGGCAACAGCCAGGAAAGCGTGACCGAACAGGATAGCAAAGATAGCACCTATAGCCTGAGCAGCACCCTGACCCTGAGCAAAGCGGATTATGAAAAACATAAAGTGTATGCGTGCGAAGTGACCCATCAGGGCCTGAGCAGCCCGGTGACCAAAAGCTTTAACCGCGGCGAATGC。

[0070] Example 3

[0071] Binding activity of human complement C5 antibody to C5

[0072] The binding activity of humanized complement C5 antibodies was assayed using an ELISA method. C5 protein was diluted to 1 μg / mL in CBS buffer and coated onto an ELISA plate at 50 μL / well. The plate was incubated at 4°C overnight. The plate was washed three times with 200 μL / well of PBST buffer. 1% BSA dissolved in PBS was then added and blocked at 37°C for 1 hour at 200 μL / well. After incubation, the plate was washed three times with 200 μL / well of PBST. The positive control, PcAb (Crovalimab, produced in-house using a Roche antibody sequence), and the humanized complement antibody were diluted to 3 μg / mL in PBST. A three-fold serial dilution was performed, and 100 μL of the aliquot was transferred to the ELISA plate using a pipette. The plate was incubated at 37°C for 1 hour. After incubation, the plate was washed three times with 200 μL / well of PBST and patted dry. Dilute HRP-labeled anti-human (or mouse) IgG antibody (all diluted 1:10000) with 1% BSA dissolved in PBS, 100 μL / well. Incubate at 37°C for 1 hour; after incubation, wash the plate 3 times with PBST, 200 μL / well, and pat dry. Add 50 μL TMB colorimetric solution to each well for 5 minutes, then add 50 μL / well hydrochloric acid to terminate the reaction. Use the absorbance mode of the enzyme reader to detect the absorbance value at 450nm (OD450). ELISA test results are as follows: Figure 1 shown.

[0073] Depend on Figure 1 It can be seen that the complement C5 humanized antibodies 6E3-HwLw, 6E3-H1L1, 6E3-H2L1 and m6E3 have strong binding ability to complement C5 protein. The EC50 of the obtained antibodies are: 6E3-HwLw = 0.039 μg / mL, 6E3-H1L1 = 0.019 μg / mL, 6E3-H2L1 = 0.020 μg / mL, Crovalimab = 0.058 μg / mL.

[0074] Example 4

[0075] Ability of 6E3-H1L1 antibody to block activation of the classical complement pathway (CP)

[0076] Human IgG was aggregated by heating to form HAIgG, an activator of the classical complement pathway. Standard reaction serum was used as a positive control for classical complement activation. An in vitro assay was established to evaluate the effect of antibodies on complement activity, using the terminal complement activation marker sC5b-9 as a detection indicator. After three freeze-thaw cycles, the cC5b-9 concentration in the standard reaction serum remained unchanged, demonstrating the stability of the serum sample. The experimental steps were briefly as follows: Plasma from healthy individuals was rapidly thawed in a 37°C water bath and immediately placed on ice. A negative control (addition of control antibody #A7001, Beyotime Biosciences), a positive control (addition of HAIgG), and an experimental group (test antibody, 6E3-H1L1, Crovalimab) were set up. 100 μL / well of mixed serum was added to each group. Samples were incubated in a 37°C water bath for 1 hour (mixed every 10 minutes). After incubation, sC5b-9 in the samples was quantified using the sC5b-9 detection kit.

[0077] From the results, it can be seen that after adding HAIgG to activate complement, the sC5b-9 concentration in the negative control group did not increase, while the sC5b-9 concentration in the positive control group increased significantly. 6E3-H1L1 and Crovalimab both have an inhibitory effect on the production of sC5b-9 activated by the classical pathway ( Figure 2 ), and the inhibitory effect of 6E3-H1L1 was better than that of Crovalimab.

[0078] Example 5

[0079] Ability of the 6E3-H1L1 antibody to block activation of the alternative complement pathway (AP)

[0080] Zymosan has the ability to activate the alternative complement pathway both in vitro and in vivo. Zymosan was added to act as an activator of the alternative complement pathway, and a standard reaction serum was used as a positive control for activation of the alternative complement pathway. An in vitro assay for evaluating antibody activation of the alternative complement pathway was established, using the terminal complement activation marker sC5b-9 as a detection indicator. After three freeze-thaw cycles, the concentration of cC5b-9 in the standard reaction serum remained unchanged, demonstrating the stability of the serum sample. The experimental steps were briefly as follows: Plasma from healthy individuals was rapidly thawed in a 37°C water bath and immediately placed on ice. A negative control (with control antibody #A7001, Beyotime Biotechnology Co., Ltd.), a positive control (with zymosan), and an experimental group (with the test antibody, 6E3-H1L1, Crovalimab) were set up. 100 μL of mixed serum was added to each well. Samples were incubated in a 37°C water bath for 1 hour, with mixing every 10 minutes. After the incubation, the sC5b-9 in the sample was quantitatively analyzed according to the method of the sC5b-9 detection kit.

[0081] From the results, it can be seen that after adding Zymosan to activate the alternative complement pathway, the sC5b-9 concentration in the negative control group did not increase, while the sC5b-9 concentration in the positive control group increased significantly. 6E3-H1L1 and Crovalimab both had an inhibitory effect on the production of sC5b-9 activated by the classical pathway ( Figure 3 ), and the inhibitory effect of 6E3-H1L1 was better than that of Crovalimab.

[0082] Example 6

[0083] C5 humanized antibody-mediated cell killing effect (CD16-NFκB-Luciferase reporter gene)

[0084] We constructed 293T-CD16 cells and then introduced the NFκB-Luciferase reporter gene vector to obtain the CD16-NFκB-Luciferase cell line. The Luciferase assay was used to detect the activation ability of the humanized C5 antibody on the 293T-CD16 cell signaling pathway. In the experiment, the weaker the detected fluorescence intensity, the stronger the activation activity of the corresponding C5 humanized antibody on the signaling pathway, which also means that the antibody-mediated cell killing effect (ADCC) activity is stronger. 293T-CD16 cells were collected and the number of cells was 2.0×10 450 μL / well of each sample was added to a 96-well plate. A negative control (PBS), a control antibody (human IgG1 antibody, #A7001, Beyotime Biosciences), a humanized C5 antibody, and a control antibody, Crovalimab, were added at the designated concentrations. Antibodies were diluted in a 1:3 series starting at 15 μg / mL. DMEM complete medium (containing 10% FBS) was used as the diluent. 50 μL / well was added to the corresponding wells. A blank control was treated with DMEM complete medium. Each sample was plated in duplicate. The 96-well plate was incubated in a 37°C, 5% CO2 incubator for 6 hours. After incubation, the 96-well plate was removed and 50 μL / well of luciferase assay reagent, equilibrated to room temperature, was added. The plate was incubated at room temperature for approximately 3 minutes. Chemiluminescent signals were detected using a multi-function microplate reader in the luminescence mode. Data were processed and histograms were generated using GraphPad Prism 8.0 data processing software. The experimental results showed that C5 humanized antibodies 6E7-HwLw, 6E3-H1L1, 6E3-H2L1 and the control antibody all showed weak luminescence signals, indicating weak ADCC activity ( Figure 4 There was little difference in ADCC effect between antibodies.

[0085] The applicant states that while the present invention uses the aforementioned embodiments to illustrate a monoclonal antibody targeting human complement C5 and its applications, the present invention is not limited to these embodiments, nor does it necessarily rely on these embodiments for implementation. Those skilled in the art will appreciate that any improvements to the present invention, equivalent substitutions for raw materials in the present invention, addition of auxiliary components, and selection of specific methods, etc., fall within the scope of protection and disclosure of the present invention.

[0086] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the scope of protection of the present invention.

[0087] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any appropriate manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A monoclonal antibody targeting human complement C5, characterized in that: The monoclonal antibody comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3; The amino acid sequence of the HCDR1 includes the sequence shown in SEQ ID NO.1, the amino acid sequence of the HCDR2 includes the sequence shown in SEQ ID NO.2, and the amino acid sequence of the HCDR3 includes the sequence shown in SEQ ID NO.3; The amino acid sequence of the LCDR1 includes the sequence shown in SEQ ID NO.4, the amino acid sequence of the LCDR2 includes the sequence shown in SEQ ID NO.5, and the amino acid sequence of the LCDR3 includes the sequence shown in SEQ ID NO.

6.

2. The monoclonal antibody targeting human complement C5 according to claim 1, characterized in that The amino acid sequence of the heavy chain variable region includes the sequence shown in SEQ ID NO.

7.

3. The monoclonal antibody targeting human complement C5 according to claim 1 or 2, characterized in that The amino acid sequence of the light chain variable region includes the sequence shown in SEQ ID NO.

8.

4. A nucleic acid molecule, characterized in that The nucleic acid molecule encodes the monoclonal antibody targeting human complement C5 according to any one of claims 1 to 3.

5. The nucleic acid molecule according to claim 4, characterized in that The nucleotide sequence of the nucleic acid molecule includes the sequence shown in SEQ ID NO.9 and the sequence shown in SEQ ID NO.

10.

6. An expression vector, characterized in that The expression vector contains the nucleic acid molecule according to claim 5.

7. A host cell, characterized in that The host cell contains the expression vector according to claim 6.

8. Use of the monoclonal antibody targeting human complement C5 according to any one of claims 1 to 3, the expression vector according to claim 6, or the host cell according to claim 7 in the preparation of a detection reagent, wherein the target of the detection reagent is complement C5.

9. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the monoclonal antibody targeting human complement C5 according to any one of claims 1 to 3, the expression vector according to claim 6, or the host cell according to claim 7, and a pharmaceutically acceptable carrier and / or diluent.

10. Use of any one or a combination of at least two of the monoclonal antibody targeting human complement C5 according to any one of claims 1 to 3, the expression vector according to claim 6, the host cell according to claim 7, or the pharmaceutical composition according to claim 9 in the preparation of drugs for preventing, improving, and treating rare diseases, autoimmune diseases, chronic kidney disease, etc., or drugs for inhibiting immune rejection.

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