Anti-IL-5 antibodies and their applications

Abstract

The present invention belongs to the field of biomedicine and discloses anti-IL-5 antibodies and their applications. The complementary determining regions H_CDR1, H_CDR2 and H_CDR3 of the heavy chain variable region of the anti-IL-5 antibody are SGYWV, SISYSGTTYYNPSLKS and GIPMDS, respectively; the complementary determining regions L_CDR1, L_CDR2 and L_CDR3 of the light chain variable region are RASESVDSYGNSFMH, RASNLES and QQSNEDPYT, respectively. The IL-5 antibodies of some examples of the present invention have undergone humanized modification, maintaining their affinity and proliferation blocking effect, and have a significant ability to block the binding of IL-5 to IL-5Rα, with an affinity greater than 10 ‑11 M, and effectively reduced immune side effects.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to anti-IL-5 antibodies and applications thereof. Background Art

[0002] Severe eosinophilic asthma (SEA) is a difficult-to-treat respiratory disease with poor clinical control. Eosinophils are crucial in the pathogenesis of SEA, causing airway epithelial damage and bronchial remodeling. Eosinophils are associated with a higher frequency of exacerbations, which reduces lung function. The current treatment for asthma is inhaled corticosteroids, but long-term use can lead to osteoporosis and hypertension. Biologic agents have shown promising results in treating severe asthma. Type 2 inflammation occurs in 50% to 70% of asthma patients. Th2 cell-mediated production of inflammatory cytokines such as IL-5, IL-4, and IL-13 increases EOS, IgE, and exhaled nitric oxide (FeNO). Therefore, reducing eosinophil counts by inhibiting proinflammatory cytokines is a highly beneficial treatment option for SEA.

[0003] IL-5 is a homodimeric cytokine linked by interchain disulfide bonds. It is crucial for regulating the proliferation, activation, and maturation of eosinophils, promoting their release from the bone marrow into the circulation. While Th2 lymphocytes are the primary source of IL-5 production, mast cells, type II innate lymphocytes (ILC2s), natural killer T cells, and even eosinophils also express and produce IL-5. IL-5Rα is an IL-5 binding receptor expressed on the surface of mature eosinophils and basophils. The intermediate complex formed by IL-5 binding to IL-5Rα recruits the β common (βc) receptor (CD131) to form a ternary complex, which then activates the JAK / STAT signaling cascade, promoting the proliferation of eosinophils (EOS). Currently available anti-IL-5 antibodies include mepolizumab and reslizumab, which neutralize IL-5 levels in vivo, thereby reducing eosinophil levels, inhibiting the differentiation and activation of EOS, and ultimately reducing the rate of asthma exacerbations. In clinical trials, antibodies targeting the IL-5 / IL-5Rα complex have been shown to reduce the rate of severe asthma exacerbations, improve forced expiratory volume in 1 second (FEV1), and reduce the use of oral corticosteroids, among other indications. Furthermore, because EOS is associated with mucus formation, reducing the number of airway eosinophils can subsequently reduce the formation of airway obstruction, thereby improving lung function. Anti-IL-5 antibodies have been shown to be effective as an emergency treatment for steroid-refractory acute severe eosinophilic asthma. In addition, the approved IL-5-targeting antibody mepolizmab has been approved for the treatment of granulomatosis with polyangiitis, eosinophilic granuloma, chronic rhinosinusitis with nasal polyposis, and Churg-Strauss syndrome.

[0004] Currently available monoclonal antibodies targeting IL-5 are all imported, expensive, and limited in accessibility. Therefore, developing a novel anti-IL-5 monoclonal antibody for injection has promising clinical application prospects and is expected to break the foreign monopoly and benefit more patients. Summary of the Invention

[0005] The object of the present invention is to overcome at least one deficiency of the prior art and provide an anti-IL-5 antibody and its use.

[0006] The technical solution adopted by the present invention is:

[0007] The first aspect of the present invention provides:

[0008] An anti-IL-5 antibody comprises a heavy chain and a light chain, wherein the heavy chain variable region comprises three complementarity determining regions H_CDR1, H_CDR2, and H_CDR3, and the light chain variable region comprises three complementarity determining regions L_CDR1, L_CDR2, and L_CDR3, and the amino acid sequences of the complementarity determining regions are as follows:

[0009] H_CDR1: SGYWV (SEQ ID NO. 1), H_CDR2: SISYSGTTYYNPSLKS (SEQ ID NO. 2) and H_CDR3: GIPMDS (SEQ ID NO. 3);

[0010] L_CDR1: RASESVDSYGNSFMH (SEQ ID NO. 4), L_CDR2: RASNLES (SEQ ID NO. 5), and L_CDR3: QQSNEDPYT (SEQ ID NO. 6).

[0011] In some examples of anti-IL-5 antibodies, the framework region of the heavy chain variable region is a humanized framework region.

[0012] In some examples of anti-IL-5 antibodies, the amino acid sequence of the heavy chain variable region is selected from one of the following:

[0013] MIL-A97-H, the amino acid sequence of which is shown in SEQ ID NO. 7;

[0014] VH1, the amino acid sequence of which is shown in SEQ ID NO. 9;

[0015] VH2, the amino acid sequence of which is shown in SEQ ID NO. 10;

[0016] VH3, the amino acid sequence of which is shown in SEQ ID NO. 11;

[0017] VH4, the amino acid sequence of which is shown in SEQ ID NO. 12;

[0018] VH5, the amino acid sequence of which is shown in SEQ ID NO. 13;

[0019] VH6, the amino acid sequence of which is shown in SEQ ID NO. 14;

[0020] The amino acid sequence of the light chain variable region is selected from the following, and its amino acid sequence is shown in SEQ ID NO. 8, 15 to 18;

[0021] MIL-A97-L, the amino acid sequence of which is shown in SEQ ID NO. 8;

[0022] VL1, the amino acid sequence of which is shown in SEQ ID NO. 15;

[0023] VL2, the amino acid sequence of which is shown in SEQ ID NO. 16;

[0024] VL3, the amino acid sequence of which is shown in SEQ ID NO. 17;

[0025] VL4, the amino acid sequence of which is shown in SEQ ID NO. 18.

[0026] In some examples of anti-IL-5 antibodies, the amino acid sequence combinations of their heavy chain variable regions and light chain variable regions are: MIL-A97-H+MIL-A97-L, VH1+VL1, VH1+VL2, VH1+VL3, VH1+VL4, VH2+VL2, VH2+VL3, VH2+VL4, VH3+VL2, VH3+VL3, VH3+VL4, VH4+VL2, VH4+VL3, VH4+VL4, VH5+VL3, VH5+VL4, VH6+VL3, VH6+VL4.

[0027] In some examples of anti-IL-5 antibodies, the antibody is a monoclonal antibody, a Fab antibody, a single domain antibody, or a multivalent antibody. Specific antibodies can be designed by combining existing technologies.

[0028] The second aspect of the present invention provides:

[0029] The first aspect of the present invention relates to the use of the anti-IL-5 antibody in the preparation of a medicament for treating IL-5-related respiratory diseases.

[0030] In some application examples, the drug is an injection or an inhalant.

[0031] In some application examples, the IL-5-related respiratory disease is selected from severe eosinophilic asthma, granulomatosis with polyangiitis, eosinophilic granuloma, chronic sinusitis with nasal polyps, Churg-Strauss syndrome, and pulmonary eosinophilia.

[0032] The third aspect of the present invention provides:

[0033] The first aspect of the present invention relates to the use of the anti-IL-5 antibody in the preparation of an IL-5 detection kit.

[0034] A fourth aspect of the present invention provides:

[0035] A gene encoding the anti-IL-5 antibody according to the first aspect of the present invention.

[0036] The specific gene sequence can be designed and synthesized using existing methods.

[0037] The beneficial effects of the present invention are:

[0038] The IL-5 antibodies of some embodiments of the present invention have been humanized and have maintained their affinity and proliferation blocking effect, and have a significant ability to block the binding of IL-5 to IL-5Rα, with an affinity greater than 10 -11 M, and effectively reduced immune side effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 This is the result of the antibody recombinant supernatant binding to human IL-5.

[0040] Figure 2 It is the result of recombinant supernatant ELISA blocking test.

[0041] Figure 3 It is the result of monoclonal antibody FACS blocking test.

[0042] Figure 4 This is the result of the monoclonal antibody TF-1 cell proliferation inhibition test.

[0043] Figure 5 These are the results of epitope analysis of hu39D10 (P33964) and MIL-A97 (P41791).

[0044] Figure 6 These are the results of epitope analysis of Mepolizumab (P33965) and MIL-A97 (P41791).

[0045] Figure 7 It is the affinity test result.

[0046] Figure 8 and Figure 9This is the result of FACS blocking test of MIL-A97 humanized antibody.

[0047] Figures 10 to 14 These are the results of TF-1 proliferation inhibition assays using different MIL-A97 humanized antibodies. DETAILED DESCRIPTION

[0048] The technical solution of the present invention is further illustrated below with reference to examples.

[0049] Example 1: Mouse immunization

[0050] BalB / C mice were grouped and immunized with proteins A (hIL-5-hFc), B (hIL-5-his), C (cyIL-5-His), and D (hIL-5-hFc+cyIL-5-His). After four rounds of routine immunization, serum was collected for titer detection. ELISA plates were then coated with IL-5-hFc, hIL-5-his, and cyIL-5-His as antigens and tested separately. All mice were then boosted with immunizations.

[0051] Example 2: Construction of mouse immune library

[0052] RNA was extracted from the spleen tissue of each mouse in Example 1 using the Trizol method. Reverse transcription was performed using the TaKaRa reverse transcription kit. PCR amplification was performed using heavy and light chain primers using cDNA as a template. A Fab phage display library was constructed using existing methods.

[0053] Example 3: Antibody Screening

[0054] The experiment used the magnetic bead method and the immune tube method to screen the mouse immune library. According to a certain inoculation amount, the mouse immune phage library in Example 2 was inoculated into 2YT (C + -T + ) medium, and cultured with shaking until OD 600 ≈0.5, add Helper Phage and infect for 1 h to 1.5 h, then centrifuge and discard the supernatant, pour into 2YT (C + -K + ) overnight in culture medium. The next day, centrifuge and remove the supernatant. Add PEG6000, mix thoroughly, and place on ice for precipitation. Centrifuge 1 hour later, discard the supernatant, add pre-chilled PBS, and pipette to dissolve the phage suspension.

[0055] 3.1 Magnetic bead screening

[0056] Magnetic bead screening is based on biotin labeling of the antigen protein (human IL-5 or monkey IL-5) and then binding it to magnetic beads coupled with streptavidin. The selection process involves incubating the antigen-bound magnetic beads with the antibody gene phage display library, washing, and eluting.

[0057] The mouse immune library suspension was diluted with 2.5% BSA and blocked for negative selection (the positive selection antigen was Fc-tagged, so negative selection protein was added during incubation at 10-fold the amount of positive selection protein). Phages were collected after negative selection. Binding and washing were performed according to the Kingfisher magnetic bead selection system protocol, and the phages were finally eluted with trypsin. The eluted phages were then used to infect SS320 cells in the logarithmic growth phase. The resulting phages were used for the next round of panning. After 3-4 rounds of panning, a pool of phage specific for IL-5 was enriched.

[0058] 3.2 Immunotube screening

[0059] Immunotube screening involves coating the antigen protein (human IL-5 or monkey IL-5) on the surface of an immunotube with high adsorption capacity, adding the phage-displayed antibody library to the immunotube, and incubating, washing, and eluting the antigen protein adsorbed on the surface of the immunotube.

[0060] 3.3 Monoclonal selection

[0061] The clone pool with good enrichment effect was selected for monoclonal plating, and the monoclonal supernatant Fab expression was induced. The ELISA method was used to detect the binding signal of monoclonal supernatant with IL-5 using IL-5 as the antigen. A total of 555 positive clones were obtained. The positive clones were selected for sequencing analysis, and a total of 50 positive antibodies with unique sequences were obtained. Through sequence similarity analysis and modification analysis, 46 antibodies were finally selected for recombinant expression.

[0062] Example 4: Recombinant expression of monoclonal antibodies and detection of IL-5 protein binding activity

[0063] The heavy chain of 46 Fab sequences was constructed on a pcDNA3.4 vector containing an Fc tag to obtain a recombinant plasmid expressing the complete heavy chain. The light chain was constructed on a pcDNA3.4 vector to obtain a recombinant plasmid expressing the complete light chain. The recombinant plasmids were recombinantly expressed in expi CHO-S cells, and the supernatant was collected for ELISA binding assay.

[0064] Human IL-5-His was diluted to 2 μg / mL with PBS and added to the ELISA plate at 30 μL per well. The plate was coated overnight, washed five times with PBST, and blocked with 5% milk at 37°C for 1 h. A gradient dilution of the recombinant antibody was added and the plate was incubated at 37°C for 1 h. The plate was washed five times with PBST, and the secondary antibody Anti-human-IgG-Fc-HRP (diluted 1:10,000 with PBS) was added and incubated at 37°C for 1 h. The plate was then washed five times with PBST, and 30 μL of TMB colorimetric solution was added to each well. The plate was reacted at room temperature in the dark for 5 min, and then 2 M sulfuric acid was added to terminate the reaction. The absorbance was measured at 450 nm using a microplate reader.

[0065] Figure 1 The binding of some antibody supernatants to human IL-5 is shown, with IPI and milk as negative controls.

[0066] Example 5: ELISA blocking activity detection of recombinant supernatant

[0067] Dilute hIL-5-hFc to 2 μg / mL in PBS, add 30 μL to an ELISA plate, and incubate overnight at 4°C. Wash the plate three times with PBST and block with 5% milk at room temperature for 2 h. Wash the plate three times with PBST, dilute the recombinant antibody to be tested in 1% milk, add 30 μL per well to the ELISA plate, and incubate at room temperature for 1 h. Without washing, add hIL-5Rα-cHis (4 μg / mL) and incubate at room temperature for 1 h. Wash the plate three times with PBST, add 6× his-HRP (Proteintech; HRP-66005) at a 1:4000 dilution, and incubate at room temperature for 1 h. After washing, add 30 μL of TMB colorimetric solution to each well and incubate at room temperature for 5 min in the dark. The reaction is terminated by adding 50 μL of 2 M sulfuric acid, and the absorbance is measured at 450 nm using a microplate reader.

[0068] Figure 2 The results show that some antibody supernatants blocked the binding of IL-5 to IL-5R at the ELISA level. The negative controls are IPI and milk. IPI was not successfully fitted, and milk is single-point data.

[0069] Example 6: Antibody purification and blocking activity detection

[0070] The cell supernatant with blocking effect in Example 5 was purified using a protein A purification column and its purity was analyzed by SDS-PAGE. Referring to the method of Example 5, the purified monoclonal antibody was retested by ELISA blocking assay.

[0071] A CHO-S stable cell line co-expressing IL-5Rα and CD131 proteins was constructed. CHO-S-IL-5Rα-CD131 cells were collected by centrifugation, washed with FACS staining buffer, and resuspended to prepare a cell suspension. The cell density was adjusted to 1×10 6 cells / mL, centrifuge and discard the supernatant. Refer to the method of patent CN116606375A for FACS blocking test. The results are as follows Figure 3 As shown, MIL-A97 antibody can block the binding of IL-5 to cell surface IL-5Rα at the FACS level. Figure 3 The data show the blocking effect of some antibodies on the binding of IL-5 to IL-5R at the FACS level, with hIL-5-mFc and cell only as single-point data.

[0072] Example 7: Proliferation inhibition assay of recombinant antibody TF-1

[0073] The recombinant antibody MIL-A97 was used to detect the proliferation inhibition of TF-1. The specific detection method was carried out according to the method disclosed in CN116606375A. The results are shown in Figure 2. Figure 4 As shown, MIL-A97 can significantly inhibit the proliferation of the TF-1-IL-5Rα-A17 cell line, and the blocking effect of MIL-A97 is comparable to that of the positive control hu39D10. The amino acid sequences of the heavy chain variable region and light chain variable region of the MIL-A97 antibody are as follows:

[0074] MIL-A97 heavy chain variable region: QVQLKESGPSLVKPSQTLSLTCSVTGDSVT- SGYW -WIRKFPGNKLEYMG- SISYSGTTYYNPSLKS -RISITRDTSMNQYYLQLNSVTTEDTATYYCAG- GIPMDS -WGQGTSVTVSS (SEQ ID NO. 7)

[0075] MIL-A97 light chain variable region: DIVLTQSPASLAVSLGQRATISC- RASESVDSYGNSFMH -WYQQKPGQPPKLLIY- RASNLES -GIPARFSGSGSRTDFTLTINPVEADDVATYYC- QQSNEDPYT -FGGGTKLEIK (SEQ ID NO. 8).

[0076] In the above amino acid sequence, the italicized regions are the CDR regions determined according to the Kabat database.

[0077] Example 8: Antibody epitope analysis

[0078] 8.1 Epitope Competition Analysis with hu39D10

[0079] hu39D10 is Reslizumab.

[0080] Human IL-5-His was diluted to 1 μg / mL in PBS and added to an ELISA plate (30 μL per well) for overnight incubation at 4°C. After washing the plate three times, the plate was blocked with 5% skim milk for 2 hours at room temperature. The antibody to be analyzed was diluted in 1% milk and added to the plate for 1 hour at room temperature. 0.5 μg / mL hu39D10-Biotin was added to the plate (30 μL per well) for 1 hour at room temperature. The plate was washed three times with PBST and NeutrAvidin-HRP (Thermo Fisher; 31001) diluted 1:2000 was added and incubated for 1 hour at room temperature. After washing, 30 μL of TMB colorimetric solution was added to each well and the plate was incubated at room temperature for 5 minutes in the dark. The reaction was terminated with 50 μL of 2 M sulfuric acid and the absorbance was measured at 450 nm using a microplate reader.

[0081] The results are as follows Figure 5 As shown, there is an epitope competition relationship between hu39D10 (P33964) and MIL-A97 (P41791).

[0082] 8.2 Epitope Competition Analysis between Candidate Antibodies and Mepolizumab

[0083] Human IL-5-His was diluted to 4 μg / mL in PBS and added to an ELISA plate at 30 μL per well for overnight incubation at 4°C. After washing the plate three times, the plate was blocked with 5% skim milk powder for 2 hours at room temperature. The antibody to be analyzed was diluted in 1% milk and added to the plate for 1 hour at room temperature. Mepolizumab-Biotin-Biotin (0.5 μg / mL) was added to the plate at 30 μL per well for 1 hour at room temperature. The plate was washed three times with PBST and NeutrAvidin-HRP (ThermoFisher; 31001) diluted 1:2000 was added for 1 hour at room temperature. After washing, 30 μL of TMB colorimetric solution was added to each well and the plate was incubated in the dark for 5 minutes at room temperature. The reaction was terminated with 50 μL of 2 M sulfuric acid and the absorbance was measured at 450 nm using a microplate reader.

[0084] The results are as follows Figure 6 As shown, MIL-A97 (P41791) and Mepolizumab (P33965) are not the same epitope.

[0085] Example 9: ELISA-based detection of the binding activity of candidate antibodies to monkey IL-5 protein and mouse IL-5 protein

[0086] Monkey IL-5-His, rat IL-5-His, and mouse IL-5-His proteins were coated on ELISA plates, and MIL-A97 (P41791) was added for binding activity detection. The specific method was described in Example 4. The results are shown in Table 1.

[0087] Table 1: Results of the binding activity test between MIL-A97 and IL-5 from different animals (EC 50 )

[0088]

[0089] The results showed that MIL-A97 has cross-reactivity between humans, mice and monkeys.

[0090] Example 10: Antibody affinity determination

[0091] Antibody affinity was determined using Biacore T200 (Cytiva). Protein A chips were used to capture the antibodies to be tested, and different concentrations of antigen IL-5-His were added for affinity testing. The results are shown in Figure 2. Figure 7 and as shown in Table 2.

[0092] Table 2: Antibody affinity determination

[0093]

[0094] The results show that the affinity of MIL-A97 is comparable to that of the positive control hu39D10.

[0095] Example 11: ELISA binding and blocking assay of MIL-A97 humanized antibody

[0096] Based on the affinity blocking activity test results, the MIL-A97 antibody (named A97-mVH-A97-mVL) was selected for humanization. The germline with the highest similarity to MIL-A97 was selected by IgBlast, the CDR region of MIL-A97 was transplanted, and different sites were selected for back mutation. The sequence was synthesized and constructed into an expression vector. The sequence is shown in Table 3 and expressed in expiCHO-S cells. Purification was performed using a protein A protein purification column. ELISA binding and ELISA blocking activity tests were performed according to the methods of Examples 4-5. The results are shown in Table 4.

[0097] Table 3: Amino acid sequences of the heavy and light chain variable regions of the humanized MIL-A97 antibody

[0098]

[0099] Note: In the table, VH refers to heavy chain and VL refers to light chain.

[0100] Table 4: ELISA activity test results of different humanized monoclonal antibodies against different IL-5 / EC 50 (μg / mL)

[0101]

[0102] Note: In the table, VH1 to VH6 refer to the heavy chains with the same numbers in Table 3, and VL1 to VL4 refer to the light chains with the same numbers in Table 3.

[0103] Example 12: FACS blocking assay of MIL-A97 humanized antibody

[0104] The FACS blocking test of the humanized antibody was performed with reference to the method of Example 6. The results are as follows: Figure 8 and Figure 9 As shown (hIL-5-mFc is single point data), A97-huVH1-A97-huVL1, A97-huVH1-A97-huVL2, A97-huVH1-A97-huVL3, A97-huVH2-A97-huVL2, A97-huVH3-A97-huVL2, A97-huVH3-A97-huVL3, huVH4-A97-huVL2, and A97-huVH4-A97-huVL3 have better blocking effects.

[0105] Example 13: Affinity detection of MIL-A97 humanized antibody

[0106] Humanized antibodies with good ELISA binding, blocking, and FACS blocking abilities were selected for affinity testing. Affinity testing of the humanized antibodies was performed using the method described in Example 10. The results, as shown in Table 5, showed that the affinity of the candidate humanized antibodies was comparable to that of the parent antibody.

[0107] Table 5 Affinity detection of humanized antibodies

[0108] .

[0109] Example 14: MIL-A97 humanized antibody TF-1 proliferation inhibition assay

[0110] Humanized antibodies were used to detect TF-1 proliferation inhibition. The specific detection method refers to patent CN116606375A. The results are as follows Figure 10-14 As shown (IgG1 and Cell+IL-5-hFc are single-point data), the candidate humanized antibodies all have the effect of inhibiting TF-1 cell proliferation.

[0111] The above is a further detailed description of the present invention and should not be considered as a limitation on the specific implementation of the present invention. For those skilled in the art, simple deductions or substitutions that do not depart from the concept of the present invention are within the scope of protection of the present invention.

Claims

1. An anti-IL-5 antibody comprising a heavy chain and a light chain, wherein the heavy chain variable region comprises three complementarity determining regions H_CDR1, H_CDR2, and H_CDR3, and the light chain variable region comprises three complementarity determining regions L_CDR1, L_CDR2, and L_CDR3, characterized in that: The amino acid sequence of the complementary determining region is as follows: H_CDR1:SGYWV H_CDR2:SISYSGTTYYNPSLKS H_CDR3:GIPMDS L_CDR1:RASESVDSYGNSFMH L_CDR2:RASNLES L_CDR3:QQSNEDPYT.

2. The anti-IL-5 antibody according to claim 1, characterized in that The framework region of the heavy chain variable region is a humanized framework region.

3. The anti-IL-5 antibody according to claim 1, wherein The amino acid sequence of the heavy chain variable region is selected from the following: MIL-A97-H: QVQLKESGPSLVKPSQTLSLTCSVTGDSVTSGYWVWIRKFPGNKLEYMGSISYSGTTYYNPSLKSRISITRDTSMNQYYLQLNSVTTEDTATYYCAGGIPMDSWGQGTSVTVSS VH1: QVQLKESGPGLVKPSQTLSLTCSVTGDSVTSGYWVWIRKHPGKGLEYMGSISYSGTTYYNPSLKSRITISRDTSMNQYYLKLSSVTAADTAVYYCAGGIPMDSWGQGTTVTVSS VH2: QVQLQESGPGLVKPSQTLSLTCSVTGDSVTSGYWVWIRQHPGKGLEYMGSISYSGTTYYNPSLKSRITISRDTSMNQYSLKLSSVTAADTAVYYCAGGIPMDSWGQGTTVTVSS VH3: QVQLQESGPGLVKPSQTLSLTCSVTGDSVTSGYWVWIRQHPGKGLEYMGSISYSGTTYYNPSLKSRVTISRDTSKNQYSLKLSSVTAADTAVYYCAGGIPMDSWGQGTTVTVSS VH4: QVQLQESGPGLVKPSQTLSLTCTVSGDSVTSGYWVWIRQHPGKGLEYMGSISYSGTTYYNPSLKSRVTISRDTSKNQYSLKLSSVTAADTAVYYCAGGIPMDSWGQGTTVTVSS VH5: QVQLQESGPGLVKPSQTLSLTCTVSGDSVTSGYWVWIRQHPGKGLEYIGSISYSGTTYYNPSLKSRVTISRDTSKNQFSLKLSSVTAADTAVYYCAGGIPMDSWGQGTTVTVSS VH6: QVQLQESGPGLVKPSQTLSLTCTVSGDSISSGYWVWIRQHPGKGLEYIGSISYSGTTYYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAGGIPMDSWGQGTTVTVSS; The amino acid sequence of the light chain variable region is selected from the following: MIL-A97-L: DIVLTQSPASLAVSLGQRATISCRASESVDSYGNSFMHWYQQKPGQPPKLLIYRASNLESGIPARFSGSGSRTDFTLTINPVEADDVATYYCQQSNEDPYTFGGGTKLEIK VL1: DIQLTQSPSSLSVSVGDRATISCRASESVDSYGNSFMHWYQQKPGQPPKLLIYRASNLESGIPARFSGSGSRTDFTLTISSVQAEDFATYYCQQSNEDPYTFGGGTKLEIK VL2: DIQLTQSPSSLSVSVGDRATITCRASESVDSYGNSFMHWYQQKPGKAPKLLIYRASNLESGIPARFSGSGSRTDFTLTISSLQPEDFATYYCQQSNEDPYTFGGGTKLEIK VL3: DIQLTQSPSSSLSASVGDRVTITCRASESVDSYGNSFMHWYQQKPGKAPKLLIYRASNLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSNEDPYTFGGGTKLEIK VL4: DIQMTQSPSSSLSASVGDRVTITCRASESVDSYGNSFMHWYQQKPGKAPKLLIYRASNLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSNEDPYTFGGGTKLEIK.

4. The anti-IL-5 antibody according to claim 3, characterized in that The amino acid sequence combination of its heavy chain variable region and light chain variable region is: MIL-A97-H+MIL-A97-L, VH1+VL1, VH1+VL2, VH1+VL3, VH1+VL4, VH2+VL2, VH2+VL3, VH2+VL4, VH3+VL2, VH3+VL3, VH3+VL4, VH4+VL2, VH4+VL3, VH4+VL4, VH5+VL3, VH5+VL4, VH6+VL3, VH6+VL4.

5. The anti-IL-5 antibody according to claim 1, wherein The antibody is a monoclonal antibody.

6. The anti-IL-5 antibody according to claim 1, characterized in that The antibody is a Fab antibody.

7. The anti-IL-5 antibody according to claim 1, characterized in that The antibody is a multivalent antibody.

8. Use of the anti-IL-5 antibody according to any one of claims 1 to 7 in the preparation of an IL-5 detection kit.

9. A gene encoding the anti-IL-5 antibody according to any one of claims 1 to 7.

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