Anti-angptl3 humanized monoclonal antibody and use thereof

Abstract

Provided are an anti-ANGPTL3 humanized monoclonal antibody and a use thereof. An original antibody is used to perform humanized transformation, and the prepared anti-ANGPTL3 humanized monoclonal antibody has higher affinity than the original antibody, indicating that the humanized antibody has high affinity with an antigen and exhibits good human-mouse cross-species reactivity, showing a good treatment effect on a mouse model of kidney disease; and the humanized monoclonal antibody has the characteristic of long half-life period, can be stably present in a mouse body for a long time, and has high safety.

Description

Anti-angptl3 humanized monoclonal antibody and application thereof TECHNICAL FIELD

[0001] The present application belongs to the field of biotechnology and antibody engineering pharmaceutical, and specifically relates to an anti-angptl3 humanized monoclonal antibody and application thereof. BACKGROUND

[0002] Angiopoietin-like protein 3(ANGPTL3) is a secreted protein, which is normally expressed at low levels in the kidney and is significantly increased in various kidney pathological conditions. It consists of an amino-terminal CCD and a carboxy-terminal FLD in its molecular structure. ANGPTL3-CCD can inhibit lipoprotein lipase, which is the functional domain of ANGPTL3 in lipid metabolism regulation(T.M.Teslovich, K.Musunuru, A.V.Smith et al. Biological, clinical and population relevance of 95 loci for blood lipids[J]. Nature, 2010, 466(7307): 707-713). ANGPTL3 also has the function of promoting angiogenesis, which is realized through ANGPTL3-FLD domain with integrin avB3 as the receptor(A.Angelakopoulou, T.Shah, R, Sofat et al. Comparative analysis of genome-wide association studies signals for lipids, diabetes, and coronary heart disease: Cardiovascular Biomarker Genetics Collaboration[J]. Eur Heart J, 2012, 33(3): 393-407.). In terms of the role of ANGPTL3 in the kidney, the expression level of ANGPTL3 in the kidney is significantly increased in patients with various kidney diseases and animal models of kidney disease(S.Romeo, W.Yin, J.Kozlitina et al. Rare loss-of-function mutations in ANGPTL family members contribute to plasma triglyceride levels in humans[I. J Clin Invest, 2009, 119(1): 70-79).At present, the research of ANGPTL3 and kidney injury is mostly focused on non-diabetic patients. Studies have found that serum ANGPTL3 levels in patients with hyperlipidemia-related proteinuria are positively correlated with 24-hour urinary protein quantification. ANGPTL3 is considered to be the connecting point of hyperlipidemia and proteinuria, and may be closely related to the generation of proteinuria associated with lipid metabolism disorders in chronic kidney disease (Yu Cai-Guo, Yuan Sha-Sha, Yang Long-Yan, et al. Angiopoietin-like 3 Is a Potential B-iomarker for Retinopathy in Type 2 Diabetic Patients [J]. Am J Ophthalmol, 2018, 191: 34-41). It is well known that podocytes cover the outside of the glomerular basement membrane, and they are an important barrier to prevent protein loss (Zhong P, Gu X, Cheng R, et al. αvβ3 integrin-targeted micellar mertansine prodrug effectively inhibits triple-negative breast cancer in vivo [J]. Int J Nanomedicine, 2017, 12: 7913-7921.). There is a small amount of ANGPTL3 deposition in normal kidney tissue, but ANGPTL3 has different degrees of increased expression in the glomerulus and renal tubules of children with primary nephrotic syndrome. The expression is significantly increased in children with minimal change nephrotic syndrome (a pathological type that highlights podocyte damage) and is positively correlated with the degree of proteinuria, so it is believed that ANGPTL3 is related to the generation of proteinuria through damage to podocytes (Luo Fei, Wu Panyun, Chen Jingfei, et al. ANGPTL3 possibly promotes cardiac angiogenesis through improving proangiogenic ability of endothelial progenitor cells after myocardial infarction [J]. Lipids Health Dis, 2018, 17(1): 184.).

[0003] Anti-ANGPTL3 monoclonal antibodies can specifically block or interfere with the activity of circulating ANGPTL3 (Dewey FE, Gusarova V, Dunbar RL, et al. Genetic and Pharmacologic Inactivation of ANGPTL3 and Cardiovascular Disease [J]. N Engl J Med, 2017, 377(3): 211-221.), and after injection of molecular targeted drugs Evinacumab and Vupanorsen, the triglyceride and cholesterol levels of dyslipidemia healthy volunteers can be dose-dependently reduced, and it is considered that ANGPTL3 can be used as a potential target for treating mixed hyperlipidemia.

[0004] The purpose of humanization of the mouse monoclonal antibody is to reduce its immunogenicity in the human body while ensuring that the binding site and affinity of the monoclonal antibody remain essentially unchanged, and to facilitate clinical transformation. The modification strategy is to transplant the complementarity determining region (CDR) sequence in the variable region, which is essential for antibody binding, into a framework composed of human sequences, and to replace the constant region as a whole with human sequences. The light chain adopts human kappa light chain constant region, and the heavy chain adopts human IgG1 heavy chain constant region.

[0005] SUMMARY

[0006] The present application utilizes the original antibody for humanization, and the modified antibody has higher affinity and high safety compared with the original antibody. Based on this, the present application is completed.

[0007] In a first aspect, the present application provides an anti-ANGPTL3 humanized monoclonal antibody, which comprises a heavy chain variable region (VH) and a light chain variable region (VL), the heavy chain variable region sequence is shown as SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 7 or SEQ ID NO. 9, and the light chain variable region sequence is shown as SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8 or SEQ ID NO. 10.

[0008] Further, when the heavy chain variable region amino acid sequence is as shown in SEQ ID NO. 3, the light chain variable region amino acid sequence is as shown in SEQ ID NO. 4; when the heavy chain variable region amino acid sequence is as shown in SEQ ID NO. 5, the light chain variable region amino acid sequence is as shown in SEQ ID NO. 6; when the heavy chain variable region amino acid sequence is as shown in SEQ ID NO. 7, the light chain variable region amino acid sequence is as shown in SEQ ID NO. 8; when the heavy chain variable region amino acid sequence is as shown in SEQ ID NO. 9, the light chain variable region amino acid sequence is as shown in SEQ ID NO. 10.

[0009] Further, the anti-ANGPTL3 humanized monoclonal antibody also includes a derivative thereof.

[0010] In a second aspect, the present application provides a nucleic acid molecule encoding the anti-ANGPTL3 humanized monoclonal antibody according to the first aspect of the present application.

[0011] In a third aspect, the present application provides an antibody conjugate comprising the monoclonal antibody according to the first aspect of the present application.

[0012] In a fourth aspect, the present application provides a composition comprising the anti-ANGPTL3 humanized monoclonal antibody according to the present application or a pharmaceutically acceptable carrier thereof.

[0013] In a fifth aspect, the present application provides use of the monoclonal antibody according to the first aspect of the present application, the antibody conjugate according to the third aspect of the present application or the composition according to the fourth aspect of the present application in the preparation of a medicament for preventing and / or treating a kidney disease or a metabolic related disease.

[0014] Further, the kidney disease includes but is not limited to various kidney diseases accompanied by proteinuria and podocyte injury such as minimal change disease, focal segmental glomerulosclerosis, diabetic nephropathy, IgA nephropathy, Henoch-Schonlein purpura nephritis, lupus nephritis, etc.

[0015] Further, the metabolic related disease includes but is not limited to diabetes, hypercholesterolemia, hypertriglyceridemia and other dyslipidemia diseases.

[0016] In a sixth aspect, the present application provides use of an agent for detecting an ANGPTL3 protein, wherein the agent comprises the anti-ANGPTL3 humanized monoclonal antibody.

[0017] In a seventh aspect, the present application provides a kit for detecting an ANGPTL3 protein, wherein the kit comprises an instruction and a detection agent, and the detection agent is the agent according to the sixth aspect of the present application. Advantages

[0018] The prepared anti-ANGPTL3 humanized monoclonal antibody has higher affinity than the original antibody, which indicates that the humanized antibody has high affinity with the antigen, has good human-mouse cross reactivity, and shows good therapeutic effect on a mouse model of kidney disease; the humanized monoclonal antibody has the characteristics of long half-life, can stably exist in the mouse body for a long time, and has high safety. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 shows the affinity ratio results of the humanized monoclonal antibody and the mouse-derived monoclonal antibody to the antigen ANGPTL3.

[0020] Figure 2 shows the SDS-PAGE results of the humanized monoclonal antibody.

[0021] Figure 3 shows the SEC-HPLC results of the humanized monoclonal antibody.

[0022] Figure 4 shows the DLS detection results of the humanized monoclonal antibody.

[0023] Figure 5 shows the SPR detection results of the humanized monoclonal antibody.

[0024] Figure 6 shows the body weight and biochemical index results of the in vivo therapeutic effect verification experiment of the humanized monoclonal antibody.

[0025] Figure 7 shows the kidney tissue pathological results of the in vivo therapeutic effect verification experiment of the humanized monoclonal antibody.

[0026] Figure 8 shows the blood concentration-time curve of the humanized monoclonal antibody in mice.

[0027] Figure 9 shows the pharmacokinetic parameters of the humanized monoclonal antibody in mice.

[0028] Figure 10 shows the body weight changes of the humanized mouse safety evaluation.

[0029] Figure 11 shows the organ index of the humanized mouse safety evaluation.

[0030] Figure 12 shows the blood routine results of the humanized mouse safety evaluation.

[0031] Figure 13 shows the biochemical index results of the humanized mouse safety evaluation.

[0032] Figure 14 shows the main organ tissue pathological results of the humanized mouse safety evaluation. DETAILED DESCRIPTION

[0033] The specific embodiments of the present application are further described below. It should be noted that the description of these embodiments is used to help understand the present application, but does not constitute a limitation on the present application. In addition, the technical features involved in the following described embodiments can be combined with each other as long as they do not conflict with each other.

[0034] The term "SDS-PAGE" refers to sodium dodecyl sulfate polyacrylamide gel electrophoresis, which is the most commonly used technique for protein expression analysis in polyacrylamide gel electrophoresis. The principle of this technique is to separate proteins in the sample according to their molecular weight. SDS-PAGE is an essential operation in the expression and purification of foreign proteins in eukaryotic cell systems, and is usually used to detect protein expression (expression level, expression distribution), and to analyze the purity of the target protein, etc.

[0035] The term "SEC-HPLC" refers to a chromatographic separation technique, also known as gel filtration chromatography. It is a method of separating molecules according to their particle size, and is commonly used for large molecules or macromolecular complexes such as proteins and polymers. SEC-HPLC has become the gold standard for assessing the purity and aggregation of recombinant proteins, and can quickly detect the purity, repeatability and batch stability of proteins.

[0036] The term "DLS" refers to Dynamic Light Scattering, which is an effective method for measuring the size and distribution of submicron and nanometer particles. The particles in the dispersion system are in a state of random motion due to the collision of solvent molecules, which is Brownian motion. DLS measures the Brownian motion of particles in the dispersion system to determine the hydrodynamic size of the particles. The rate of Brownian motion can be quantified by the translational diffusion coefficient, usually represented by D. In a dispersion system, smaller particles diffuse faster and larger particles diffuse slower. Since the measured particles may have different shapes and the particle surface may be combined with other substances such as ions or polymers, we generally equate the shape of the measured particles to a sphere with the same diffusion rate, and the diameter of the sphere is taken as the hydrodynamic size of the particle.

[0037] The term "SPR" is used herein: SPR detection technology is a new detection and analysis technology based on optics and label-free, which can be used to dynamically monitor the whole process of interaction between biomolecules. SPR technology has the characteristics of high throughput, high flexibility and high sensitivity, which can allow researchers to characterize the interaction of biomolecules in binding studies, and can be widely used for other different molecules such as ions, molecular fragments, small molecules, proteins, viruses, etc. SPR is to detect the interaction between ligands and analytes on a biosensor chip, and then to detect the properties and structures of substances. In this way, we can analyze various organic or inorganic substances in the sample in real time and accurately, and infer the binding constant Ka, dissociation constant Kd, affinity constant KD and binding kinetics between molecules.

[0038] The experimental methods in the following examples are all conventional methods unless otherwise specified. The experimental materials used in the following examples are all commercially available unless otherwise specified.

[0039] Example 1 Humanization modification

[0040] According to the provided original antibody sequence information, the homology model of the antibody is obtained by modeling, and the CDRs are analyzed The framework amino acids in the range usually affect the conformation or antigen binding activity of the CDRs. The human germline is obtained by IMGT analysis, and the selected human germline framework is spliced with the CDRs of the antibody. The framework region sequence of the designed humanized antibody is compared with that of the original antibody. By analyzing the homology modeling results of the parent antibody, the amino acids similar to the surface residues of the human antibody are selected for replacement on the basis of maintaining the activity of the antibody and reducing heterogeneity, and the humanized antibody sequence is designed.

[0041] The sequence of the variable region of the original antibody is as follows:

[0042] The sequence of the variable region of the heavy chain (SEQ ID NO. 1) is as follows:

[0043] The sequence of the variable region of the light chain (SEQ ID NO. 2) is as follows:

[0044] The specific operation is as follows:

[0045] (1) Homology modeling of parent antibody and determination of key amino acids in FR region

[0046] A. Determine the variable region and the complementarity determining region (CDR);

[0047] B. Search for homologous proteins of antibody variable region, determine template protein structure and sequence;

[0048] C. Structural similarity analysis of template proteins to determine the structural conserved regions (SCR) and random loop regions (Loop) of the template proteins;

[0049] D. Building the SCR spatial structure, searching and analyzing the Loop conformation, and building and obtaining the initial conformation of antibody VH and VL from scratch;

[0050] E. Further optimization of the initial conformation, first adjusting the dihedral angle of the model conformation, then optimizing the antibody variable region conformation through steps such as steepest descent, conjugate gradient, Newton mechanics under CVFF, Gromos force field, and finally obtaining a stable spatial conformation through dynamics optimization, and evaluating the obtained antibody variable region conformation;

[0051] F. Selecting the same template containing VH and VL conformation as a reference, considering the interaction mode of VH and VL, analyzing the accessible surface and apparent electrostatic distribution, and obtaining the antibody V region conformation;

[0052] G. Investigating antigen-antibody interaction, predicting the binding mode and affinity of the two, and thus performing antigen epitope prediction, antibody humanization, and affinity modification;

[0053] H. Calculating the accessible surface area and apparent electrostatic distribution to determine the important amino acid residues in the antibody FR region.

[0054] (2) Acquisition of human FR region template, CDR grafting, and computer structure simulation analysis

[0055] A. Retrieving mouse antibody V region homologous sequences in the human source library, and comparing and inducing high-frequency amino acid residues and positions in the human antibody FR;

[0056] B. According to the results of accessible surface area, apparent electrostatic distribution, and sequence alignment, determine the amino acid residues that need to be changed during humanization;

[0057] C. Simulate the V region conformation of the humanized antibody, and compare the conformation of the parent antibody and the humanized antibody through structure superposition;

[0058] D. Simulate and optimize the structure of the antibody-antigen complex after humanization, and evaluate the biological activity of the heterologous antibody after humanization through interaction energy, intermolecular hydrogen bonds, and recognition epitopes.

[0059] According to the above humanization modification process, four pairs of humanized light and heavy chain variable region sequences are designed, wherein the four pairs of humanized light and heavy chain variable region sequences are as follows:

[0060] Heavy chain variable region sequence (SEQ ID NO. 3):

[0061] Light chain variable region sequence (SEQ ID NO. 4):

[0062] Heavy chain variable region sequence (SEQ ID NO. 5):

[0063] Light chain variable region sequence (SEQ ID NO. 6)

[0064] Heavy chain variable region sequence (SEQ ID NO. 7):

[0065] Light chain variable region sequence (SEQ ID NO. 8):

[0066] Heavy chain variable region sequence (SEQ ID NO. 9):

[0067] Light chain variable region sequence (SEQ ID NO. 10):

[0068] Example 2 Affinity comparison of humanized mAb and pre-engineered murine mAb to antigen ANGPTL3

[0069] The affinity of recombinantly expressed humanized mAb and murine mAb to antigen ANGPTL3 was measured and compared using the SPR method, and the results showed that the affinity constant K D = 1.859E-9 M for humanized mAb hAb to hANGPTL3, the affinity constant K D = 6.728E-9 M for murine mAb mAb to hANGPTL3; the affinity constant K D = 2.066E-9 M for humanized mAb hAb to hANGPTL3-FLD, the affinity constant K D = 7.795E-9 M for murine mAb mAb to hANGPTL3-FLD; the affinity constant K D = 4.665E-9 M for humanized mAb hAb to mANGPTL3, the affinity constant K D = 1.626E-8 M for murine mAb mAb to mANGPTL3; the affinity of humanized antibody is better than that of murine antibody, whether it is for human ANGPTL3 or mouse ANGPTL3, or for ANGPTL3 full-length protein or ANGPTL3-FLD fragment.

[0070] Example 3 Antibody characterization

[0071] 3.1 Experimental methods

[0072] (1) SDS-PAGE detection of the molecular weight of the antibody: Tris-glycine gel was configured for electrophoresis, and after electrophoresis, the gel was taken out, stained, and then rinsed overnight, and then imaged, and the molecular weight was determined according to the band position.

[0073] (2) SEC-HPLC detection of the purity of the antibody: after diluting the purified protein, injecting the chromatographic column, and judging the purity of the antibody.

[0074] (3) DLS detection of the stability, particle size, and dispersion coefficient of the antibody: the polydispersity, particle size, and particle size distribution of the sample were obtained, and the Tm and Tagg values were determined to reveal the thermal stability and colloidal stability of the antibody.

[0075] (4) SPR detection of the affinity of the antibody to the antigen: human IgG capture antibodies were pre-fixed on a CM5 chip (GE Healthcare, USA), the antibody was captured on the chip, and then the antigen hANGPTL3 or mANGPTL3 was flowed through the chip, bound and dissociated, and the affinity was calculated.

[0076] 3.2 Experimental results

[0077] (1) SDS-PAGE

[0078] After the humanized monoclonal antibody was expressed and purified, SDS-PAGE electrophoresis was performed, and the molecular weight was 137.41 kDa under non-reducing conditions, and the molecular weights of the light and heavy chains were 30.28 kDa and 51.09 kDa, respectively, under reducing conditions. The protein band was clear and had good purity (Figure 2).

[0079] (2) SEC-HPLC

[0080] After the humanized monoclonal antibody was expressed and purified, SEC-HPLC detection was performed, and the main peak molecular weight was 171.8 kDa, the area percentage was 98.8%, the secondary peak molecular weight was 377.7 kDa, and the area percentage was 1.2%, suggesting that the purity of this batch of humanized monoclonal antibody was high (Figure 3).

[0081] (3) DLS

[0082] Using Uncle all-in-one protein stability analyzer, dynamic light scattering DLS technology and temperature rise experiment were used to detect the stability, particle size and particle size distribution of the humanized monoclonal antibody. The results showed that the Tm value was 62.08℃, the Tagg value was 54.09℃, the particle size was 10.99 nm, and the PDI coefficient was 0.108, indicating that the humanized monoclonal antibody had good thermal stability, colloidal stability and polydispersity (Figure 4).

[0083] (4) SPR

[0084] The affinity of the humanized monoclonal antibody and hANGPTL3-His antigen and mANGPTL3-His antigen was detected using SPR technology with a Biacore T200 instrument, and the results showed that the affinity constant of the humanized monoclonal antibody to hANGPTL3-His was KD = 4.740E-10 M, and the affinity constant to mANGPTL3-His was KD = 4.953E-10 M, indicating that the humanized antibody had high affinity to the antigen and had good human-mouse cross-reactivity (Figure 5).

[0085] Example 4 Therapeutic Effect Verification of ANGPTL3 Humanized Monoclonal Antibody

[0086] 4.1 Experimental Method

[0087] To verify the kidney protection effect of the anti-ANGPTL3 humanized monoclonal antibody on the adriamycin nephropathy mouse model, and compare the therapeutic effect with the positive drug hormone, the experimental scheme is designed as follows: male Balb / c mice are selected and randomly divided into 5 groups. The control group (Control) is injected with physiological saline 10.5 mg / kg once by tail vein, and the model group (ADR) is injected with adriamycin 10.5 mg / kg once by tail vein. The humanized monoclonal antibody group (hAb) is modeled as above, and hAb 20 mg / kg is injected intraperitoneally twice a week. The positive drug group (steroid) is modeled as above, and prednisone 2 mg / kg is administered once a day by gavage. The drug administration is started the day after modeling, and the mice are sacrificed after 3 weeks of continuous administration. The body weight, urinary albumin creatinine ratio (UACR), serum creatinine (SCR), blood urea nitrogen (BUN), triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) are detected, and the kidney tissue is stained with PAS to observe and count the mesangial proliferation.

[0088] 4.2 Experimental Results

[0089] (1) Body weight and biochemical indicators

[0090] In the in vivo therapeutic effect verification experiment of the humanized monoclonal antibody on the adriamycin nephropathy mouse model, the model group showed significant weight loss, increased urinary protein, impaired renal function, and elevated blood lipids compared to the control group. After treatment with the humanized monoclonal antibody, urinary protein was significantly reduced, kidney function was protected, and blood lipids were reduced, showing good therapeutic effect. The humanized monoclonal antibody performed significantly better than the positive drug hormone in reducing urinary protein and blood lipid levels (Figure 6).

[0091] (2) Kidney tissue pathology

[0092] In the in vivo efficacy verification experiment of humanized monoclonal antibody in adriamycin nephropathy mouse model, compared with the control group, the modeling group showed pathological changes of mesangial cell proliferation and matrix increase in the glomerular region, accompanied by focal segmental glomerulosclerosis, while the humanized monoclonal antibody treatment significantly improved this pathological damage, showing reduced cell proliferation and matrix deposition in the mesangial region, restored normal course and opening of the capillary loop, and the improvement effect was significantly better than that of the positive drug hormone (Figure 7).

[0093] Pharmacokinetics of Example 5

[0094] To clarify the pharmacokinetic characteristics of anti-ANGPTL3 humanized monoclonal antibody in mice, as a reference for the development of the administration scheme, the experimental scheme is designed as follows: male Balb / c mice are selected and randomly divided into 3 dose groups, and single tail vein injection of humanized monoclonal antibody 5 mg / kg, 10 mg / kg and 20 mg / kg is performed, respectively. Blood is taken at 15 min, 1 d, 3 d, 7 d, 14 d, 21 d and 28 d after administration, the blood drug concentration is detected, the blood drug concentration-time curve is drawn, and the pharmacokinetic parameters are calculated.

[0095] The change of blood drug concentration in 3 dose groups in mice with time is shown in the blood drug concentration-time curve (Figure 8), and the corresponding pharmacokinetic parameters are fitted and calculated. It can be seen that with the increase of the administration concentration, the drug concentration in mice increases, the elimination rate accelerates, and the half-life prolongs, and the half-life corresponding to the low to high dose is 10.4, 12.0 and 14.1 days, respectively, indicating that the humanized monoclonal antibody has the characteristics of long half-life and can exist stably in mice for a long time (Figure 9).

[0096] Safety evaluation of ANGPTL3 humanized monoclonal antibody of Example 6

[0097] To evaluate the safety of anti-ANGPTL3 humanized monoclonal antibody in mice, in order to predict possible adverse reactions in the future, the experimental scheme is designed as follows: male Balb / c mice are selected and randomly divided into 2 groups, the control group (Control) PBS and the humanized monoclonal antibody group (hAb) 50 mg / kg are injected intraperitoneally 2 times a week, and the mice are sacrificed after 4 weeks of continuous administration. The changes of body weight, organ index, blood routine, serum biochemical indicators and histopathological conditions of main parenchymal organs are monitored.

[0098] The results show that after 4 weeks of continuous administration of humanized monoclonal antibody, there is no significant change in body weight between the administration group and the control group (Figure 10), and there is no significant change in organ index except for the slight increase in liver index (Figure 11), and there is no significant difference in blood routine indexes between the two groups (Figure 12), and there is no significant difference in liver and kidney function and blood lipid level between the two groups, but the LDH and LDH1 in the heart muscle enzyme spectrum in the administration group appear significantly increased, and the CK and CK-MB appear an increasing trend, indicating the risk of myocardial injury (Figure 13), and in the aspect of histopathology, the liver of the mice in the administration group appears mild water degeneration, and the rest of the heart, spleen, lung, kidney, brain and other parenchymal organs show no significant abnormalities (Figure 14).

Claims

1. An anti-ANGPTL3 humanized monoclonal antibody, comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the heavy chain variable region sequence is as shown in SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 7 or SEQ ID NO. 9, and the light chain variable region sequence is as shown in SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 8 or SEQ ID NO.

10. 2.The monoclonal antibody of claim 1, wherein when the heavy chain variable region amino acid sequence is as shown in SEQ ID NO. 3, the light chain variable region amino acid sequence is as shown in SEQ ID NO. 4; when the heavy chain variable region amino acid sequence is as shown in SEQ ID NO. 5, the light chain variable region amino acid sequence is as shown in SEQ ID NO. 6; when the heavy chain variable region amino acid sequence is as shown in SEQ ID NO. 7, the light chain variable region amino acid sequence is as shown in SEQ ID NO. 8; and when the heavy chain variable region amino acid sequence is as shown in SEQ ID NO. 9, the light chain variable region amino acid sequence is as shown in SEQ ID NO.

10. 3.The monoclonal antibody of claim 1, wherein the anti-ANGPTL3 humanized monoclonal antibody further comprises a derivative thereof. 4.A nucleic acid molecule encoding the anti-ANGPTL3 humanized monoclonal antibody of the first aspect of the present application. 5.An antibody conjugate comprising the monoclonal antibody of claim 1. 6.A composition comprising the anti-ANGPTL3 humanized monoclonal antibody of claim 1 or a pharmaceutically acceptable carrier thereof. 7.Use of the monoclonal antibody of the first aspect of the present application, the antibody conjugate of the third aspect of the present application or the composition of the fourth aspect of the present application in the preparation of a medicament for preventing and / or treating a kidney disease or a metabolic related disease. 8.The use of claim 1, wherein the kidney disease comprises, but is not limited to, various types of kidney diseases accompanied by proteinuria and podocyte injury, such as minimal change disease, focal segmental glomerulosclerosis, diabetic nephropathy, IgA nephropathy, Henoch-Schonlein purpura nephritis, lupus nephritis, etc., and the metabolic related disease comprises, but is not limited to, diabetes, hypercholesterolemia, hypertriglyceridemia and other dyslipidemia diseases. 9.Use of an agent for detecting ANGPTL3 protein, wherein the agent comprises the anti-ANGPTL3 humanized monoclonal antibody. 10.A kit for detecting ANGPTL3 protein, comprising an instruction and a detection agent, wherein the detection agent is the agent of claim 9.

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